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User Manual LABScreen HLA Antibody Detection

1. A Zu G tari nah 237585 4753 110 PCIHC Ratio 180 3 OOF Pos 234814 476 2 118 119 0 2 2 188 PCIHC Ratio a 69 0 1 2 152 138 0 2 172 S s EIS AE Pos O76 Fos 19451 0 388 8 120 O77 Pos 3d 0 7 180 LABScreen V3 1 IVD Rev 1 pdf Appendix A LABScreen Reports 58 of 72 OLI UD 0176 022007 Mixed Data Individual Report MESF conversion values are shown only on Mixed Individual Reports e Generated by selecting LABScan 100 gt Import Reprocess Mixed gt Individual in the Data Import Report dialog Figure A 5 Mixed Data Individual Report Cateye LABScreen Mixed Data Page 1 CatIDvLot LSMT2 012 Sample ID 16030 est Date 05 26 2006 03 17 57 Tested By AW Session D X ABSCREEM DATAFILESXSMTZ 012 OUTPUT CSS default Background Values pf Overall Beads Resut Rawdata Ratio Count Class Pos 235620 590403 5 J3b38l 592260 2 420040 5640915 Ja B85 595407 0 230140 5964500 2307 0 5838933 Class Il Pos 355255 33480 b 34495 4 MICA Pos O16 194510 491273 3 017 Fos 397 0 318158 Controls MC bl f 223885 FC 1062 0 468334 5 xem LLL LABScreen V3 1 IVD Rev 1 pdf Appendix A LABScreen Reports 59 of 72 OLI UD 0176 022007 Mixed Data Summary Report Summarizes the results of all samples within a selected batch on one line per sample Generated by selecting LABScan 100 gt Import Reprocess Mixed batch then sele
2. Figure 3 7 Creating a New MESF Conversion Lot Current Heads ID WiFI 44 245 1038 Tasso 8 mm Lot Update Initial Pairs Save Save ae MyLoto22d Update default values 3 Transfer the data from the Median and SFI columns in the Quantiplex Beads Data Sheet Figure 3 6 to the Current MFI and MESF columns in the MFI to MESF Conversion Settings form Figure 3 7 Note that the Beads ID numbers are not necessarily in ascending order but the EFI values are Make sure that the order in the Conversion Settings fields is the same as in the Beads Data Sheet 4 Enter a name in the Save As Lot field and press Save As You don t need to clear the Update Default Val ues check box as this option applies only to the simple Save button directly above it 5 You can confirm the existence of the new conversion lot in the LABScreen database table by accessing the MESF Lot Selection look up Figure 3 8 Do this by clicking the search button to the right of the Lot LABScreen_V3 1_IVD Rev 1 pdf Chapter 3 Analysis 22 of 72 OLI UD 0176 022007 3 7 07 field at the top of the MFI to MESF Conversion Settings form After you have confirmed that the new record 1s there close this form and return to the main menu Figure 3 8 Lot Selection Look Up Table MESF Lot Selection E 2007012213332 FOo701LZ81426 DRAFTL O 200701291
3. LABScreen V3 1 IVD Rev 1 pdf Chapter 3 Analysis 36 of 72 OLI UD 0176 022007 3 7 07 Processing Multiple Samples Figure 3 30 Antibody Screening By Batch LABScreen Antibody Screening by batch l Serum ID to Analyze Select specific samples within the batch to analyze v Class v Class I To Printer By Session ID P Analyze ey Range From as to nn Cancel By Tech ID 185 cats Lott C By Test Date 7 CO Not Yet Analyzed cand saved You can process multiple samples after importing as follows Select Analysis gt Analyze By Batch from the Main menu to access the LABScreen Antibody Screening By Batch dialog Figure 3 30 2 Choose from among these subtractive filters to specify how you want to process the batch e By Session ID processes a batch based on the session ID e By Range processes a specified range of samples with adjacent Sample IDs e By Tech ID processes samples loaded into LABScreen by the user with the specified ID this ID is the one supplied when you Import a LABScan file it can be viewed and edited in the Maintenance gt Reading Data Entry form e By Cat Lot as stated Cat IDs and Lot can be viewed and edited in the Maintenance gt Reading gt Data Entry form e By Test Date as stated this ID can be viewed and edited in the Maintenance gt Reading gt Data Entry form e Not Yet Analyzed only those samples that have not yet been Saved Accepted by th
4. SA Analysis Report This report can be printed from the Analysis window When MEFS conversion is activated the report also contains MESF values Figure A 9 Single Antigen Analysis Report 02 16 2007 LABScreen Data Page 1 atlD Lot L81 amp 1 82 001 Session ID 4 A amp BSCREEN DATAFILES LS1A1 2 C8w Test Date 09 30 2002 05 12 05 PM Tested By A Hc 2400 Pe 5 456 0 Pe Nc 220 Sample ID 419 9003 08 30 02 fo Positive Percent 8 5 000 Percent 6 4 18 750 Percent 8 4 2 31 250 Ratio Maximum F atio 51 625 Minimum Ratio O 515 Cutoff ford 35 322 Cutoff far 4 15 818 Cutott far 2 ob Specificity 531010 B20 4350 7 0 A3362 0 A32 7 0 B8 101 00 B60 C1 0 B27 2 0 Abb E20 4330170 A251 0 D AAT Bait A23 O0 B4810 8296010 BB EOD ATT OL AL 3 B4210 B7310 Overall True Positives 25 False Positives 0 False Maegatives 4 True Maegatives 53 CREG Possible CHEG antibody s are specificity TruePas FalseMeg K val AFC z 2 0315 200 E b 343 lt Ar C gt B7 B27 B13 B40 B60 B61 B4 lt 20 C gt A10 A25 A25 A34 Abb Al A25 ABB ABS ASD A31 Ass MESF MESF Beads Lot DRAFT10 Processed on 200702161558 Y 4175 21 8 20 30519x Beads ID Ren Reagent ID Ravwdata Ratio Count Specificities 015 B P A311 14160 291697 7 31 5 175 e O19 B RBU ZUO2 13766 264103 6 51 3 174 By BWG O14 B FP A3O1 12825 2645905 48 3 FF ASO UT B P A301 104 78r 216934 0 39 6 223 A33 016 4 R A201 12079 243442 5 33 5 1838 A32 O81 4 RBS 01 117930 243635 3
5. gens Serum Information analysis results and demographic information for samples that have been Saved Accepted upon analysis in the Class I or Class II Analysis Results window Figure 3 13 Results for samples that have not yet been Saved Accepted can be accessed by selecting Analysis Serum Information HLA Typing Information accesses tables that display cell demographic information serological antibody typ ing from LABScreen analyses and DNA allele assignments from LABType analyses for samples that have been Saved Accepted Similar tables for samples that have not been Saved Accepted can be accessed by selecting Analysis HLA Typing Information Class Class Il Analysis Results the principal interfaces where analyses of Class I and Class II results are carried out See Analyzing Data p 24 LABScan 100 Raw Data accesses panels that display raw data for each tested serum including PC and NC val ues session ID and NIH scores Archive LABScreen Info performs an archiving function for LABScreen catalog information similar to that described under the File Archive and Delete Data topic Update LABScreen Info performs a retrieval function for LABScreen catalog information similar to that described under the File Retrieve Data topic Add New Lot for LABScreen Mixed a tool for loading product catalog information into the database and assigning a new lot number to the product Reports Menu Options
6. Rxn AD NWCnA OO eC Nn is the MC value for each bead n that will be provided for each lot MC iz the median value af NC bead 1 for each test PC iz the median value of PC bead 2 for each test Problematic Negative Weak Postive strong Very Strang Positive Positive Positive Cutotts ere 0 00 HA 1 5 HE 5 0 XC 10 0 HE 20 0 Symbols 0 1 J 4 G 5 Cancel Cutotts were initialized ta 1 5 5 10 20 Symbols were set to be 01 24 6 5 respectively SA Advanced Parameter Settings The SA Advanced Settings form Figure 5 3 is used to set cutoff levels and NIH score symbols for Single Antigen analyses It is accessed by selecting Update Parameters gt LABScreen SA Adv Settings from the Main menu Table 5 3 Update Parameters gt SA Advanced Settings ew ree States the formulas used to determine reac Reaction Positive Classification tion strengths in assigning NIH scores based Guidelines and Formulas p 49 on the Working Range of the reaction Cutoffs Cutoff constants used in the Reaction Classification formulas LABScreen V3 1 IVD Rev 1 pdf Chapter 5 Parameters and Algorithms 43 of 72 OLI UD 0176 022007 3 7 07 Table 5 3 Update Parameters gt SA Advanced Settings pomi Symbols NIH score symbols used the reports and data base tables By default these are 1 2 4 and 8 A bead is designated as Problematic if it does not have a reading In contrast to PRA scoring Single Antigen NIH sc
7. laboratory director or technologist trained in histocompatibility testing 1s required to review all data to detect any problems with the software Product Description One Lambda LABScreen antibody screening tests use color coded microbeads coated with purified Class I or Class II HLA antigens to detect Class I or Class II HLA antibodies in human sera Up to 100 beads may be combined in one suspension for a single test The sera to be tested are incubated with LABScreen beads HLA antibodies present in the test sera bind to the antigens and are then labeled with R Phycoerythrin PE conju gated goat anti human immunoglobulin G IgG IgG is the major circulating antibody in mammals and par ticipates in many immune responses The LABScan 100 flow analyzer detects the fluorescent emission of the phycoerythrin from each bead as it is extracted from the tray well The reaction pattern of the test serum is compared to the lot specific worksheet defining the antigen array From this comparison percent PRA and HLA specificity can be made If done by hand this comparison is a tedious and error prone undertaking When done using LABScreen Software the assignments can be made in seconds In a typical LABScan data collection session the analyzer will continue to draw beads from the tray well until the minimum bead count threshold has been reached by all the beads This minimum bead count threshold is generally set at 100 beads LABScreen products are ava
8. 1 pdf co 0 C Co OF Co OF co OF co oo co co co co co co cn Percent Percent 5 6 99 10 Percent 85 4 100 00 Percento 6 4 2 Hc a L Pe 3 096 0 00 00 100 007 Pe Nc 3 7 MESF Beads Lot 1234567890 Processed an 2007072211054 Y 1372 1835 25 03853 ME E18278 C09 E5061 0469 E5482 01 06 GEB C 4513 E12535 C0086 PU C0406 E3825 E140772 E 20366 E18401 040 E1 3066 El fe OLI UD 0176 022007 Rawdata Ratio Count 370 22245 1 0 295 24189 558653 6 31 9 137 23905 552110 6 129 2 146 23509 542975 5 98 9 150 23936 5520133 1347 140 20311 469309 5 106 3 243 23454 5417202 89 0 155 22241 5137744 9 21 8 161 24090 5563613 53 1 110 23992 554115 0 276 145 23461 541870 0 443 158 3096 0 72699 8 D 1 257 24036 555128 7 769 101 24274 5560811 9 22 8 100 23540 5437015 477 155 23853 5508026 79 0 245 21178 489284 3 55 1 186 z3405 540581 3 41 3 187 24084 556234 5 544 155 23596 544991 7 474 150 specificities Ad 2 B57 8201 BV 6 CAG10 Aq 80 B18 50 BWE CVV 6 A11 23 B49 52 Bud CUNT 12 Add 24 B27 48 Bid B CA B Add 24 654 59 BWE 4 CH A11 24 659 60 B4 6 CHI 10 A11 29 616 38 BWE 4 C2 A11 33 B51 54 Bud 6 CHE 10 A11 B38 75 BB CNT B A211 513 62 Er 6 CY E A211 B51 76 Era 6 CVO As bo B44 47 EV Cy Ass 6 B53 53 Ba CMS 14 Ass 4 Bes 44 Av G Cig iT Ass 4 BS fo Be Cv 16 As4 B56 61 BWG OVW 15 Age r4 B45 72 Be Cv CVE Ad 69 B35 48 Ave 4 CVWE T2 A
9. 123 LSIPRA 007 lt 014 gt 42 B13 Ba Cg aie Bd GiG TP 11 6 151 LS1IPRA OOF 090 A34 BS6 BB2 C4 Cag Bye lt TH 4 T 117 LSIPRA 007 020 A24 B59 B60 CAN OO Big Bie lt TN gt 4 69 149 LSIPRA DD7 022 311 A23 B49 B52 CWT C2 Ed lt TN gt 4 5 6 181 LS1PRA 007 O85 A11 B39 B75 Cii OWS EE Table 3 3 Adjust Cutoffs and Change Background Controls opin Box increments decrements all NBG Background Values by the same amount Applies value specified in the Spin Box Apply Resets BG values to Default values before Defaults any user edits Applies any changes and recalculates the NIH Update scores applies yellow alert color to any NBG ratio that is 0 25 of the cutoff between NIH scores of 1 and 2 Restore NBG Values to previous values MEN Display Cutoff Adjustment entry fields for Cutoff Weak Positive Positive Strong Positive amp Very Strong Positive PRA or Negative Grey Area amp Positive Mixed Saves changes returns to Analysis Results window Cancels returns to Analysis Results window Saves background values for current sample Save to to a BGV file Loads background values from a saved BGV Update from file for analysis of current sample LABScreen_V3 1_IVD Rev 1 pdf Chapter 3 Analysis 33 of 72 OLI UD 0176 022007 3 7 07 Table 3 3 Adjust Cutoffs and Change Background Controls Replaces the NBG values with the Raw Data Values for the sample Used when processing cm a sample as a ne
10. Specificities Table Figure 3 14 Percent Positives Panel Figure 3 15 Control Panel Figure 3 16 Chi Squares Table Figure 3 17 and Figure 3 18 CREG Display Figure 3 19 LABScreen_V3 1_IVD Rev 1 pdf Chapter 3 Analysis 25 of 72 OLI UD 0176 022007 3 7 07 Figure 3 13 Class Analysis Results Window Class I Antibody Analysis Result for 41 W4136 l C Specificitiez Average TruePas FalzePoz FalseNeq TruehMec R value Strength Chisquare Percent Positives Overall 6 667 4 5 1 40 0 699 33 33 25 8 Ad 6 571 T T 1 40 0 588 28 57 19 0 Percent 8 5 45 AT4 7 000 2 5 0 4n 0 504 50 010 11 3 Percent 8 6 25 45 Percent 8 5 4 54 55 Percent 8 5 4 7 arar 24 6 5 reactions are defined by cutotts of 1 50 5 00 10 00 20 00 times normalized background values Cat ID s Analyzect LS1PRA 007 7 Print before save Chiu Specificties TP FP FH TN R Val X E Demographics Into Ve Pun 1 i s g B T A33 2 3 2 38 O39 B 236 2 3 2 38 0 38 Clear Reset Recatc Change Parameters 332 E13 1 4 1 39 027 3 2 B18 1 4 1 3d 0 27 FpiEn cre CREG By Ag Exclude Antigens Exclude Own Antigens 1 4 1 Control saluez JLETPRA OOF NC413 5 PC7396 0 PC MCH 7 8 Background Default Possible CREG antibody s are Specificity TruePos FalseNeg R val 102 12 15 8 428 4102 gt AT Ad6 APY n23 AZH A11 Figure 3 14 Analysis Result Specificities Table
11. Updated 200702197055 v MFI to MESF conversion activate Current Heads ID MFI MESF Beadl s 2 77 Minimum RS 0 299 Head 7 255 Maximum MET 23000 Heads 57 1031 25855 Calc Head4d o7 5176 115350 urrent y 20 3228 x 4151 0809 req 9987825596 Heads g 22540 460793 Lot Update Initial Pairs Save Save ac MyLot 220 cancel Restore from Default Update default values Algorithms Calculation of the Normalized BackGround Ratio The reactivity of a test sample is calculated based on the median fluorescence values which are one of the data type in the LABScan csv file The strength of the reactivity is defined as the ratio R of the normalized sam ple median to the normalized background median The abbreviations used in this section are defined below NBG Normalized BackGround SN Median fluorescence value for Sample bead N Sync bead Sample specific median fluorescence value for Negative Control bead BGy BackGround fluorescence value for bead N is by default a predetermined back ground value from OLI reference data BGyc bead BackGround fluorescence value for Negative Control bead the negative control bead is not coated with HLA antigen and is used to determine the non specific binding for each bead e For LABScreen PRA or LABScreen Single Antigen the Normalized BackGround NBG for each bead is calculated as follows LABScreen V3 1 IVD Rev 1 pdf Chapter 5 Parameters and Algorithms 47 of 72 OLI U
12. any case LABScreen will perform Class I analy ses on each sample first LABScreen V3 1 IVD Rev 1 pdf Chapter 3 Analysis 15 of 72 OLI UD 0176 022007 3 7 07 If you are analyzing samples using a Single Antigen product that combines two or three different cata log IDs and Lot numbers see File Naming Conventions for LABScreen Products p 15 for guidelines on choosing the correct catalog ID If you specify a catalog that does not match the assay LABScreen will generate a bead conflict error message If you specify a catalog that is not used say an unnecessary Class II catalog lot when only Class I sample data are available and then re import the same batch later and specify the appropriate cata log s LABScreen will generate a data cleanup message indicating that the unusable or old catalog data is being purged from the database You can ignore the message 4 Select one or more report options Print Antigen Distribution Table with Report see Figure A 10 Typical Antigen Distribution List ing for an example Preview Report First displays a preview of the data report with results pos neg grey area for each bead for each Class The report of each sample is on a separate page See Figure A 4 Mixed Data Full Report This 1s the default Analyze After Import imports all serum data then performs the same function as when you select Analysis Analysis by Batch By Session ID which displays the Antibody Analysis Resul
13. the main menu or from interfaces accessible from the main menu A small number of tasks can be executed directly from the tool bar below the main menu LABScreen Menus File Menu File options include database record archival and deletion data exportation and database table rebuilding Archive and Delete Data tools to save four categories of data to archive files with options to physically delete records marked for deletion Archived data are saved to arf files to a user specified location Data archival is intended to help the user reduce the number of samples present at any given time in the LABScreen database thus facilitating sorting and analysis of the existing records Retrieve Data tools to retrieve the archived data as described above Export Data a utility that exports reaction data and Class I and Class II results for the specified sample IDs in dBase Excel or ASCI text format Mixed Assay data can be exported only in Excel format Print Setup this is a standard system function that requires no comment Index Rebuild this function reindexes database tables that may have become damaged as the result of a sys tem malfunction Pack Database this function packs the database to reduce storage requirements Records that have been marked for deletion are physically deleted Edit Menu The edit options access system text editing functions that require no comment They can be invoked with the keyboard combinati
14. to MESF Conversion Settings llle 45 ADO uo 2190 99 93 Ber Proh eoneeeeent ees Paucde hab aaueenatametetad ATUS 47 Calculation of the Normalized BackGround Ratio 2 0 0 ees 47 Determination of Positive Negative Cutoff 0 6 ce nes 48 Reaction Positive Classification Guidelines and Formulas 0 0000 49 Statistical and Scoring Formulas 0 0 0 0 cent eens 50 Antigen Specificity Calculation 0 0 llis 51 Appendix A LABScreen Reports LAB Aa R PO e eee e e ar EEEE EE 55 Repons MonU m m 53 Patient Antibody Summary Report 2 0 0 anrea 56 Patient Antibody Report 0 0 0 ee eee e teen e eee 56 Patient Cell Typing Report ius voa oa 5600 44 booed ta bee ath Hb a a Eta e dar eee 57 LABScreen Mixed Data Full Report 0 0 0c nanne 58 Mixed Data Individual Report llle 50 Mixed Data Summary Report 22222 04 4enseterauese Ua ERU So EUR PCI E UD SUR ER Ee 60 PRA Dala Repol 6l SA Raw Data ROPO eese seeen a EEES 62 SA Analysis Repol e oua IER CRTIRSEEERE nian RRT AEE ESPERE NES Pd 63 Antigen Distribution Listing llleleeeeeeeeeeee ers 64 Sample Raw Data Report 264e4 06 325 ups i RSASDESSd RSU EAR RSS Sad PET E EE 65 Class and Class II Panel Listings llle 66 Class and Class II Analysis Antibody Screening Reports 00000000 67 Class I and Class II Analysis Antibody
15. via the Adjust Cutoffs functions Figure 3 16 Similarly the reaction definitions reproduced in report footers reflect global and not local settings Figure 3 15 Percent Positives Panel Percent Positives Percent 8 24 09 Percent 8 6 40 00 Percent 3 6 4 49 09 Percent 5 6 4 2 65 45 2 4 6 5 reactions are defined by cutoffs of 1 50 5 00 10 00 20 00 times normalized background values e Control Panel the buttons in this panel Figure 3 16 control the analysis functions of the Analysis Results window and are discussed in some detail in Table 3 2 Analysis Window Controls A further set of controls used when adjusting cutoffs and setting background values is discussed in Table 3 3 Adjust Cutoffs and Change Background Controls Figure 3 16 Analysis Window Control Panel Iz Print before save ge x al Demographics Info Clear Reset Recatc ES adjust cutorts ll Change Parameters FoF ni CREG By ag Exclude Antigens Exclude Own antigens Control s aluez LS1TPRA OOF MC125 5 PC 3313 0 PONC 25 4 Background Default e Chi Squares Table antigens that meet the R Value and Score criteria for being classified as specific ities are displayed in the Specificities Table Figure 3 14 By default the R Value threshold is 0 25 and the Score must be greater than 2 The Score S is defined as TP FN The statistics of remainder antigens i e those antigens that display false positives an
16. 0 6e ee or ne oneal m e mea CR Ahad 24 PANY FU Dalt ccc ars oo aed Be es ba eh eae Gees aes Gnas Cee ewes Gee aed 24 Viewing Results by Antigen 0 0 ce eee teen eens 3 Douai Guidi M 34 Color Coding in Revise Cutoff Raw Data Grid 20 0 ccc es 35 Changing Parameters Locally Temporarily 0 0 0 0 cc cece eee eee ees 36 EXCINGING ANU EIS esscr ere tpa hee SR II Gach bas oe HUS bho Hae GR ees 36 Processing Multiple Samples 0 0 eee ee eens 37 Chapter 4 File and Data Maintenance wur oecesecd eee es oeue hohe EE bee cee eee aero eee eee EIRE ERASE E 39 Retrieving Archived Data 0 0 0 cc eee eee e eee eee e ees 40 LABScreen V3 1 IVD Rev 1 pdf Table of Contents iii of 72 OLI UD 0176 022007 Packing the dDaLdDOSEs aoa hoe ep cree eres Cas ERE Ra TEE Td tehendaaeens 40 Rebuilding the MGC uu acci aces ae Bead Gm teas qanm eed ae ee qu RUE Errem dg nt 40 Chapter 5 Parameters and Algorithms Updating Global Parameters llli 4 Reaction Definitions Specificity Selection Methods and Report Header Information 41 Changing the Date FOrtf alt 423 22 045 2060 040008 94506000058 ERO Pa PR RD EdPPTUERO 42 Advanced SeN IRE E ETITCETTTEZTTTLCICRCITOL TE TO DT OT TET OT LT TT 42 PRA Paraimicter ciu M EPPFTTTTPPrm rT 42 SA Advanced Parameter Settings 0 0 ccc eee 43 Mixed Advanced Parameter Settings 0 0 0 ccc ccc eee teens 44 MFI
17. 070219165 amumum D Slope Intercent RR Val 21 0542 4077 37 0 99976 9 You can use the Lot browse button to select a different calibration lot for a different Luminex machine or you can use the Updated browse button to select an updated calibration lot with a different time stamp for the same machine Figure 3 11 Figure 3 11 Different Time Stamped Versions of Conversion Record MESF Update Record Selection for Lot MyLot 220 MyLot zz 200704191650 MyLotoezO0 400701460255 Select Cancel Reprocessing a LABScan 100 File If you wish to reanalyze the same LABScan data using a different product lot usually a newer one you can reprocess a LABScan 100 file that has been previously imported Follow these instructions Instead of providing a Luminex data file name as in a regular import select the Luminex session name from the pull down 2 Specify the different product s 3 All other processing options are the same as when importing Analyzing Data If you check the Analyze After Import option when importing the LABScan data LABScreen immediately proceeds to analysis If you choose to only import the data by leaving the Analyze After Import option unchecked you will need to start up the analysis after the import 1 Onthe Main menu click Analysis and then select one of the following e Analyze by Batch use this option to process multiple samples one after another See Processing Multiple
18. 2 007 Previews Report First Full Report Individual Repot J Summary Report Use Default Background wales Negative Background Cl Use Specific ID Controls ie PC Test Beads qn 004 O06 M DTE uus O06 Oo 010 011 016 TT TT TT 4 Each normalized background values each background value minus HC background are replaced by the minimum background value when the value ix less than minimum This minimum value iz specified in LABScreen Mixed Advanced Setting Report Only Import Data i Import amp Report Exit 2 Choose the report output options e Preview Report and or one or more of the following e Full Report Class I and Class II results Overall and for each Bead for all samples in the batch four samples per page includes raw data NBG Ratio and bead count report indicates if background val ues are default specified by Sample ID or manually entered See Figure A 4 Mixed Data Full Report e Individual Report same information as in the Full Report for all samples in the batch but with each sample on a separate page See Figure A 5 Mixed Data Individual Report e Summary Report summary results for each sample on a single line report includes overall result for Class I and Class II Pos Neg Grey Area Raw Data and counts for each sample s NC and PC beads and the sample s PC NC ratio See Figure A 6 Mixed Data Summary Report Choose to include MFI to MESF conversions as d
19. 20 56 of 72 z 8 Rxn Incl 11 B4 T1 40 00 fO 00 100 0 Pos irg 55 5 DAUA ODF Rival Chsu B r 4 1 1 7 3 8 Patient Cell Typing Report Collected detailed HLA Typing for all samples linked with the specified Patient ID e Generated by selecting Report gt Patient HLA Report from the main menu Figure A 3 Patient Cell Typing Report Patient Cell Typing Report for SIMIT3413 SMITHERS WAYLON 02 02 2007 C ell ID Sample ABO HLA A HLA B HLA C HLA Bw Date HLA DRB1 HLA_DOB1 HLA DRB3 45 A1 V 1135 122 2004 B Ad BIHI 4 1 LOT 7 1 12 2 2004 A O1LOL B 7uzz ciwoso1 LABScreen_V3 1_IVD Rev 1 pdf Appendix A LABScreen Reports 57 of 72 OLI UD 0176 022007 LABScreen Mixed Data Full Report Lists the results of all samples within a selected batch Due to space restrictions MESF values are not shown on the Mixed Data Full Report e Generated by selecting LABScan 100 gt Import Reprocess Mixed batch then selecting the Full Report option in the Data Import Report dialog Figure A 4 Mixed Data Full Report 02 07 2007 LABScreen Mixed Data CatlD Lot LSM12 012 Session ID JSLABRSCREENM DATAFILESSESMT2 O12 OUTPUT Cay Test Date 05 26 2006 03 17 57 Tested By Vy default Background Values Result Rawdata Rato Count Rawdata Count H 3 5 d E d R 5030 235962 0 4711 118 Pos 242 5 10 7 290 Bl 3 O07 Pos 256590 472 7 att O70 Pos 466 0 a2 Z2 PC 1082 0 127 Pos 220040 450 0 152 S030 10 0 263
20. 3 BB CWA IE 1 arro osei teese 19425 104 25 14551 n226 B47 65 Bwa CWDS eo 4 coc c m Oo E E E os os E E E E m LABScreen V3 1 IVD Rev 1 pdf Chapter 3 Analysis 35 of 72 OLI UD 0176 022007 3 7 07 Changing Parameters Locally Temporarily You can change parameters for the current sample from the Analysis window Temporary parameter setting are not persistent Thus if you return to the sample at a later time you must reset the parameters to the desired val ues See Updating Global Parameters p 41 for details on the parameter settings Excluding Antigens To exclude antigens from the Analysis click Agn Exclude to access the Antigen Selection for Analysis form The usual selection conventions apply e Shift click to select adjacent antigens e Ctrl click to select non contiguous antigens e A button with single arrow moves selected antigens e A button with double arrow moves all antigens in the field Figure 3 29 Antigen Selection For Analysis Window Antigen Selection for Analysis Included for Excluded Always excluded the analysis temporarily a _ gt 4 _ EIE ENDE 2 Atemporary exclusion applies only to the sample currently under analysis If you return to the same sample at a later time you must reapply the temporary exclusion 3 A permanent exclusion applies to the current sample and to other samples in other batches at all times
21. 30 6 323 Be1 Bye 035 1 RBS3U1 340 5 11090 1 0 9 178 B53 Bv 4 OBS 1 RBA UT 452 0 13354 Z 0 9 2a B47 By 076 1 RBTS1U 3470 11222 1 0 9 235 B71 BWG 071 1 REBS3U1 456 0 13435 4 0 9 239 B59 Bg 065 1 RASO01 240 0 10064 7 0 5 225 B50 BG 058 1 REB33U05 200 0 9415 0 0 5 351 B39 B5 O80 1 RBS8U01 ao 3821 1 0 7 351 Bro BWG OB 1 FR EBABUT 2440 8130 7 0 6 zB B46 BG O01 ME 246 0 9211 9 1 0 S21 002 PC 3455 0 1174961 3 1 5 555 LABScreen_V3 1_IVD Rev 1 pdf OLI UD 0176 022007 Appendix A LABScreen Reports Antigen Distribution Listing The Antigen Distribution Listing shows how many beads in the product would react with a given antigen To learn the specificities of a given bead consult the Panel Listing for that product Figure A 12 Class I and Class II Antigen Distribu tion listings are similar in format A Distribution Listing can be generated when importing or reprocessing PRA data by selecting the Include Panel with Report option Figure A 10 Typical Antigen Distribution Listing Class II Antigen Distribution for LSZzPEA Lot UO Antigen Antigen Antigen LABScreen_V3 1_IVD Rev 1 pdf Appendix A LABScreen Reports 64 of 72 OLI UD 0176 022007 Sample Raw Data Report Contains raw data values statistics and specificities for the sample currently under analysis e To generate select Raw Data button from within the Class I Class II Analysis Results window or from within the Maintenance gt Readings gt R
22. 4171 00004 Z LABScreen_V3 1_IVD Rev 1 pdf Appendix A LABScreen Reports 68 of 72 OLI UD 0176 022007 Class and Class Il Analysis Antibody Screening Reports 2 x 2 Listing This report contains statistics for all the antigens in the assay including those not selected as specificities for the sample You can select Patient Antibody Report to generate analysis results reports for one or more patients with contiguous Patient IDs e Generated after selecting 2 x 2 Report output option for the specific sample in the Temporary Parameters Setting form accessible from the Analysis Window or in the Update Parameters form Figure 5 1 Global Parameter Settings Form In the detail of the report shown below only the beginning and ending antigens are included Figure A 15 Class and Class Il Antibody Screening Reports 2 x 2 Listing LABScreen Class I Antibody Screening Patient s name Serum Id Mo SUNNAMED 91077 Patient Id Face Center Id Date sampled fof Analysis ex2 Listing Date tested O4 44 2006 ALA Typing Control Values ZXLSIPPRA 0l gt NHC 3 0 PO 564 0 PC HC 183 0 Avg Specificity Score IP FP FN TN aed 6 4 5 lz l 37 CT 17 6 0 4 l3 l 37 BS 6 0 Z 15 E 38 B48 6 0 Z 15 E 38 E41 6 0 Z 15 38 B 7 amp 6 0 l l 38 Cura 2 0 l l 4 a4 A30 E 0 l l 4 a4 Aes E 0 l l 4 a4 AL 6 0 z 15 7 3l A3 E 0 l l E 33 CUTS 6 0 z 15 a8 z0 Cra 6 0 l l amp 3z LABScreen V3 1 I
23. 436 DRAFTL O z00702191054 MyLotoz19 z00702191055 i ad Im Loc zz z n702191565 0 elect Cancel 6 Select LABScan 100 gt Import MESF Bead Data to access the like named form It should now display the newly created lot in the MESF Beads Lot field Figure 3 9 Click the browse button to locate a Luminex csv file containing LABScan calibration data The filenname probably contains the characters QPLX Quantiplex or the like Figure 3 9 Importing MESF Beads Data MESF Beads Data Import Luminex Data File Tech ID TATESTOATAI 2607 LECANTOPLA OOT IDIOOSIRUNE Vari MESF Beads Lot z MyLoto220 Regression Values cline 21 0547 ies SUR menia RR Val 9 9997675 MESF Values E M o 077 7083 087 26855 067 115350 081 360723 Calc Only Import Data Exit 7 Click Import Data If you have chosen a compatible conversion file LABScreen will display a Bead Number Matched message then briefly display the linear conversion formula associated with that lot 8 When you next import any kind of session data PRA Single Antigen or Mixed Antigen note that the current MFI to MESF Conversion Lot appears in the Lot field in the upper right corner of the Import Report form Figure 3 10 LABScreen V3 1 IVD Rev 1 pdf Chapter 3 Analysis 23 of 72 OLI UD 0176 022007 3 7 07 Figure 3 10 Import Report Dialog Detail Iw MFIta MESF conversion active Lot amp MMyLoto220 Updated 99
24. AI amp 2 06A designates LS1A01 Lot 006 LS51A02 Lot 007 e LSIA2 amp 3 05B designates LS1A02 Lot 005 LS1A03 Lot 007 e LSIAI amp 2 04Z designates LS1A01 Lot 004 LSIA02 Lot 003 e LSIAI amp 2 07Y designates LS1A01 Lot 007 LS1A02 Lot 005 The Catalog IDs and Lot number combinations of three groups consist of the character C plus three numbers which designate the lot numbers for each group e LSIAI23 C557 designates LS1A01 Lot 005 LS1A02 Lot 005 LS1A03 Lot 007 e LSIAI23 C657 designates LS1A01 Lot 006 LS1A02 Lot 005 LS1A03 Lot 007 e LSIAI23 C775 designates LS1A01 Lot 007 LS1A02 Lot 007 L81A03 Lot 005 Importing PRA and Single Antigen Data The importation of PRA and Single Antigen files is almost identical Batch importation is a multi step process Figure 3 1 Click the to the right of the Luminex Data File field to open a standard Windows Find File dialog and locate the LABScan csv file 2 Enter a two character Technician ID to identify you on the analysis reports You will need this identifica tion later when you want to search for and continue the analysis on samples that you personally have loaded into LABScreen 3 Use the drop down lists to specify the appropriate catalog ID s for the analysis If carrying out both Class I and Class II analyses on the sera specify a Class I catalog in one field and a Class II in the other It does not matter which field you use for which Class In
25. Analysis 18 of 72 OLI UD 0176 022007 3 7 07 Figure 3 3 Current MFI to MESF Conversion Curve Iv MFI ta MESF conversion active Lot MvLat Updated Sloe Intercept RR val Beads IC MFI MESF Mp ss ax MAF I 8 Choose the execution mode e Report Only generates the LABScreen SA or PRA Raw Data Report without creating any records to the database e Import Only imports the data creating records in the database without generating a report e Import and Report imports the data creating records in the database and generates reports 9 When you first import the Luminex data you have the option to Analyze After Import which starts the analysis automatically upon import See Analyzing Data p 24 for details Importing Mixed Antigen Data The steps followed when importing Mixed Antigen data are largely identical to those when importing PRA and Single Antigen data Figure 3 4 There are however different reporting and background value specifica tion options 1 Just as when importing PRA or SA data e Locate the Luminex Data File e Supply your two character Technician ID e Specify the LABScreen product by Catalog ID and Lot LABScreen V3 1 IVD Rev 1 pdf Chapter 3 Analysis 19 of 72 OLI UD 0176 022007 3 7 07 Figure 3 4 Import Mixed Data LABScreen Mixed 10 17 beads format Data Import Report I Luminex Data File Tech ID JALABSCREEN DATAFILESILSM12 OOF OLITPLIT CSS 9 ww zat ID 7 Lotz 5h1
26. Class I and Class II antigens Import MESF Beads Data import MESF Molecules of Equivalent Soluble Fluorophores beads data This data is used to generate MFI to MESF conversion formulas Import Reprocess LABScreen Mixed 4 bead format Data the 4 bead assay type is an obsolete product line However if you do have 4 beads data that you wish to revisit the application can be configured to import reprocess 4 bead csv files as follows 1 Exit the LABScreen application 2 Assuming that LABScreen has been installed to the default C location navigate to the C labscrn myflags folder This folder contains a number of flag files that instruct the application to ignore data for obsolete products 3 Locate the file NO LSM ABEADS FLG and rename it to LSM_4BEADS FLG When you next launch LABScreen the four bead Import Reprocess options will be enabled Analysis Menu The Analysis options provide access to the LABScreen analysis functions The options are briefly described below See Chapter 3 Analysis for usage details Serum Information provides access to Class I and Class II Analysis Details specificities tables as well as Class I and Class II Antibody Histories draw test reaction data and product tested against for the samples in the database ordered by Serum ID These tables contain data for all samples that have been imported into the LABScreen database A similar set of Serum Information tables can be accessed by sele
27. D 0176 022007 3 7 07 BGy Eq 5 1 NBG Syr NC BGyc The Normalized BackGround Ratio NBG Ratio for each bead 1s the ratio of the median raw data value for each bead Sy to its Normalized BackGround NBG S Ss Eq 5 2 NBGRatio e uius Hen GL NBG BGy BGyc e For LABScreen Mixed a subtractive normalization is used in which the normalized median is defined as the median value of the Class I or Class II coated bead minus the median value of the NC bead The NBG ratio is thus S sS Eq 5 3 NBGRatio 1N BGy BGyc Determination of Positive Negative Cutoff ForLABScreen PRA and Mixed 1 1 Select the NBG ratio that gives a significant shift over background fluorescent value when the back ground value is obtained using the negative serum in 3 5 replicate tests If you prefer test 5 10 serum samples from non transfused non transplanted male donors to obtain an average background value 1 2 Validate the cut off using 5 10 reference alloserum samples with defined HLA antibody specificity The NGB ratio values for the expected positive antigen reactions should be above the cut off 1 3 Additional positive negative reactions may be noted If necessary adjust the LABScreen assay cut off to match the sensitivity of a previously accepted antibody detection assay 2 For LABScreen Single Antigen 2 1 Test negative control serum or several negative serum samples by following Step 1 1 above 2 2 Define Working Range WR
28. Evi 6 CVS 10 018 43 32 650 56 BWG Cy B O87 423 68 B49 60 Beg 6 CVV 10 O70 433 36 B53 63 Eu Cyd 14 035 43 30 635 55 BWE CAM 45 033 230 80 B42 72 BYE CWTA 025 423 66 B41 71 BWE CWE 17 5 003 41 80 616 50 Bye C2 65 089 424 31 856 59 Bg B CWA 45 023 42 30 B27 44 By CV B4 0723 Ad 30 B42 71 EVE CVV O 17 016 A24 29 E3761 Bg B CVE 6 0113 423 66 B37 72 Big 6 CWZ 015 A2 80 B75 B4 6 CVV 8 003 42 29 B57 64 Bd 6 CVE 18 019 22 25 618 65 BWE CVVB 12 4 x 088 A24 25 B18 76 Be CVV9 12 8 O07 43 33 6838 EV 6 CWWT 12 4 o 028 432 68 844 47 Bev CNT OTF a2 tT B13 Figure 3 28 Color Coding in Revise Cutoff Raw Data Grid Cp E ze e t0a7a 11453 11455 9025 4335 beso 8WAECWTI2 pes fe aso 1sei 244 94 267 6 2789 21330 M134 85552 BW OWA Jorr fi 165 000 12 31 609 48 BW CWT B T oss 15330 15939 430 53 B41 73 BS CIS 126 13 41133 B51 54 Bid 6 CI 10 119 64 130 76 A228 BST B4 Bid 6 CVV8 18 176 74 A2 66 BE3 B5 Ei 6 CVV8 16 E a g 13344 145 84 145 84 A228 B7 46 EB Cu 15 242 43 264 94 264 94 A2 B46 BF BAB CAM T 7 a E x a Aa Cer ix a Fi exi ca 3 en o Tay a oy co oy Dani olijolololola joielaeialilejmaela eielejejeje jae c m hJ c J 168 1 A 26 B51 64 Bild 6 CUWB 14 151 85 A33 3B B53 53 Eia Cia 14 125500 1 000 11483 12550 12550 x 100 00 342 62 arasa arasa 29536 8336 B455
29. H score of 8 positive LABScreen V3 1 IVD Rev 1 pdf Chapter 5 Parameters and Algorithms 49 of 72 OLI UD 0176 022007 3 7 07 Mixed Antigen Formulas e NBG Ratio lt 1 2 Negative e NBG Ratio between 1 2 and 1 5 Grey area e NBG Ratio above 1 5 Positive Statistical and Scoring Formulas TPs FPs TNs and FNs True Positive TP False Positive FP True Negative TN and False Negative FN are defined as shown in Table 5 6 For example if an antigen is present but there no serum reaction detected the reading would be considered a False Negative To make the formulas more concise the cell designations A B C D are used in place of TP FN FP and TN Table 5 6 TP FP TN and FN Reaction Designations LL Antigen Absent FP C TN D R Value the Correlation Coefficient R is a measurement of the interdependence of two random variables R ranges in value from 1 to 1 indicating perfect negative correlation at 1 absence of correlation at zero and perfect positive correlation at 1 R Value The R Value 1s calculated using the formula E AD DC Eq 5 6 ACA B A C B D C D Values range from 1 to 1 Chi Square Chi Square the Chi Square estimate of confidence xe is calculated using the formula CN N AD DC Eon A B A C B D C D Values range from 0 to infinity Score Score the Score S is the difference TP FN The higher this number the stronger the corre
30. LESILSTPRA_OO CSW Cat Ib 1 Cat ID 2 L52PRA Oo one C Print Antigen Distribution Table with Report C Analyze non prampt mode to printer Previews Report First Q Analyze after Import L Use Default Background Values Negative Background from ID A25 Waar 5 Report Only Import Data Import amp Report Exit LABScreen V3 1 IVD Rev 1 pdf Chapter 3 Analysis 17 of 72 OLI UD 0176 022007 3 7 07 6 Accept the default background values option These are the OLI defaults for the product Occasionally you may wish to specify an alternative set of background values from a specific tray well in the batch This means that all analyses for the entire batch would use those data values as background val ues To specify an alternative set of background values using data values from one well of the batch e Clear the Use Default Background Values check box e Open the batch s LABScan csv file in Excel and locate the column containing the Sample IDs LABScreen will use the data values in the row corresponding to the data type which LABScreen has been set up to use This is typically the median Figure 3 2 Batch Data File in Excel sample 2 3 A W413b 413 5 38b ar A x5 328 7 B45 7643 5 AJS Who S 186 577 5 b226 ASU Wil 315 5 fal Hd AJb WJA 2 bood 332 AJ4 Whi A6 309 5096 5 2024 A31 pgg bha BUbS 5240 5 AJ WSA 144 bado P3 e Copy the Sample ID from the csv file into the Negative Background fro
31. Panel of the current sample Saves any manual edits to the analysis and Figure 3 12 LABScreen Anti Accepts the sample analysis Saved Accepted body Screening before Sample samples appear in the Maintenance gt Class Selection Il Results tables before being Saved Accepted the samples appear only in the Analysis gt Serum Information tables Exits the Analysis Results window without saving accepting the sample analysis results when doing batch analyses moves to the next analysis or sample Performs and displays Chi Sq analyses for Figure 3 17 Chi Square Table remainder antigens antigens not included in for Remainder Antigens the Specificities Table if the CRGE options is selected the Chi Sq tables shows the CREGs of which the remainder antigens are members Toggle for the Chi Sq button above hiding the Chi Sq display Prints analysis report right click previews the Figure A 13 Class and Class ll report Antibody Screening Reports Prints raw data report right click previews the Figure A 11 Sample Raw Data report Report Inverts NIH scores 8s become 1s 6s become Figure 3 14 Analysis Result 2S 2s become 6s and 1s become 8s LAB Specificities Table Screen recalculates the statistical analysis using the inverted scores thus testing for the absence of antigens rather than their pres ence Toggles with the button above restoring the original NIH scores Accesses the Sample d
32. Samples p 57 e Analyze Class I use this and the following options to analyze one sample at a time e Analyze Class II e Analyze Class I and Class II LABScreen V3 1 IVD Rev 1 pdf Chapter 3 Analysis 24 of 72 OLI UD 0176 022007 3 7 07 The three single sample options invoke the LABScreen Antibody Screening window Figure 3 12 When first accessed it will not be populated with values Later if you decide to analyze another sample this window will show the results of the just completed analysis Figure 3 12 LABScreen Antibody Screening before Sample Selection LABScreen Antibody Screening i x C Update Demographics Serum ID to Analyze II 2 Antibody Result for sample Date rf Marne Speciticties Average TruePos FaleePos Fale Neg TrueNeg R value Strength Chi quare Overall Selected Cat ID s 2 Select a Sample ID by clicking on the question mark next to the Sample ID field Figure 3 12 and selecting an ID from the pick list If you check the Update Demographics option the program will open the Sample Information demo graphic information form before performing the analysis The demographics form can also be accessed from other points in the program 3 The analysis starts as soon as you select a Sample ID or finish updating the demographic information The analysis results appear in the full Analysis Results window Figure 3 13 which contains five panels Each panel will be discussed separately
33. Screening Reports Reversed 68 Class I and Class II Analysis Antibody Screening Reports 2 x 2 Listing 69 Changing Report Paper 3176 26 gaan edamaryecdononedbecouereeedouekeodeawereyeeun 70 Index LABScreen_V3 1_IVD Rev 1 pdf Table of Contents iv of 72 OLI UD 0176 022007 Chapter 1 Introduction Welcome to LABScreen Software LABScreen Software is an HLA screening application that aids users of One Lambda products with identifying the presence or absence of specific antibodies in an individual s serum sample This software imports data from the LABScan 100 analyzer for the purpose of suggesting HLA antibody specificities LABScreen Software can generate a large variety of reports and is designed to store large amounts of patient demographic data and analysis result from previous testing to aid in the understanding of a patient s histocompatibility status Disclaimer All One Lambda software products are designed to assist personnel experienced in HLA analysis by suggest ing typing results However any clinical or diagnostic results must be carefully reviewed by a person qualified in HLA typing to assure correctness The software may be used to aid in suggesting results but should not be used as the sole method for determining reportable results The software is meant as a laboratory aid not as a source of definitive results The software design does not mitigate hazards associated with the software The
34. User Manual LABScreen HLA Antibody Detection Software LABScreen v 3 1 2007 02 For In Vitro Diagnostic Use Mb ONE LAMBDA INC 20001 Kittridge Street Canoga Park CA 91303 2801 Advancing Transplant Diagnostics Tel 818 702 0042 Fax 818 702 6904 www onelambda com Manual Template H1 LABScreen V3 1 IVD Rev 1 pdf supersedes LABScreen V2 7 IVD Rev O pdf OLI UD 0176 022007 3 7 07 Copyright 2007 One Lambda Inc LABScreen is a registered trademark of One Lambda Inc LABScan 100 is a trademark of One Lambda Inc 91 uminex is a registered trademark of Luminex Corporation Windows is a registered trademark of Microsoft Corporation All One Lambda software products are designed to assist personnel experienced in HLA analysis by suggesting typing results However any clinical or diagnostic results must be carefully reviewed by a person qualified in HLA typing to assure correctness The software may be used to aid in suggesting results but should not be used as the sole method for determining reportable results The software is meant as a laboratory aid not as a source of definitive results The software design does not mitigate hazards associated with the software The laboratory director or technologist trained in histocompatibility testing is required to review all data to detect any problems with the software Table of Contents Chapter 1 Introduction Irc e EEEE E cibdenehuceeagtaneee eed eanneeaedaueds ec dauc
35. VD Rev 1 pdf OLI UD 0176 022007 Appendix A LABScreen Reports Background Dbefault 55 55 55 55 BS BS SS 5E SS 5E SS 5E SS oOo oOo A A A d Percent positive 30 913 Total valid reactions 55 Mumber of positives 1 Positive reactions are 6 5 Overall strength index 5 858 5 Percent 8 1 52 Percent 28 6 30 51 Percent 5 6 4 70 914 Percent 5 6 4 2 100 004 E SCIT Chi Fisher s Value Incl Index Scare Tail 397 53 33 0 nu 5 67 0 008536 336 sO nu Oo 00 Geel O 0z 7777 Z930 100 00 Oo 00 4_64 0 091582 Z930 100 00 Oo 00 4_64 0 091582 Z930 100 00 Oo 00 4_64 0 091582 203 100 00 D 00 2 20 D 303091 075 20 00 100 00 O 31 0 6702700 075 0 00 Oo 00 O 31 0 670200 075 20 00 Oo 00 O 31 0 670200 083 E EE Oo 00 O 38 D 704755 108 16 67 Oo 00 O 64 0 653710 111 20 00 Oo 00 0 68 0 481952 Lae 14 25 Oo 00 1 04 O 416754 69 of 72 Changing Report Paper Size You can change the report paper size from the default U S letter size to A 4 as follows 1 2 3 4 Exit the LABScreen application Assuming that LABScreen has been installed in the default C location navigate to C labscrn Sort the files by type and locate the two MS DOS batch files A4_size_report and Letter_size_report Launch the A4 size report batch file This replaces the contents of the folder C labsern Reports with the contents of the folder C labscrn REPORTS_A4 The next time you generate a LABScreen rep
36. WR NBG pax NBG pin Eq 5 4 2 3 Define cut off points within the Working Range Relative Cutoff X WR NBG pax NBG min NBG pin Eq 5 5 where X user defined percent cut off point within the Working Range for negative 1 gray area 2 weak positive 4 or strong positive 8 The cut off points are relative to the maximum and minimum NBG ratios Therefore the definition of Negative Weak Positive and Strong Positive will change for every sample 2 4 Set criteria to define positive vs negative reactions for example where R is the reading value e If the ratio of Rmay Rmin gt 8 then apply the calculation in Step 2 3 If the serum test value is greater than a given cut off assign the corresponding NIH score for that particular bead reaction LABScreen V3 1 IVD Rev 1 pdf Chapter 5 Parameters and Algorithms 48 of 72 OLI UD 0176 022007 3 7 07 e If the ratio of Rmax Rmin lt 8 AND If Rmax gt 5 then Rmin should be adjusted to one half of the Rmax and the relative NBG ratio cutoff should be re computed based on the adjusted Rmin The reaction is then scored as above If Rmax lt 5 then the reaction of the test serum with that bead is negative Assign a score of 1 Reaction Positive Classification Guidelines and Formulas General parameters for LABScreen assays Bead counts should not be lower than 50 events per bead Patient samples with high background may be due to e Hydrophobicity of the plastic or the anti
37. a listing of the currently defined member antigens in Class I CREGs Cross Reacting Groups The modification of CREG definitions 1s done in the Public Antigen Maintenance Data Entry form accessible via the main menu Maintenance options LABScreen V3 1 IVD Rev 1 pdf Appendix A LABScreen Reports 55 of 72 OLI UD 0176 022007 Patient Antibody Summary Report Collected typing results for all samples linked with the specified Patient ID showing only percent positive reactions and specificities e Generated by selecting Report gt Patient Antibody Summary Report from the main menu Figure A 1 Patient Antibody Summary Report MARKOV SOREN Cat lD Lot Test Date LSZPR A OOF 05 1 820604 LS1PRA OOF Der 272004 Baro are yz Patient Sera Report for MARK5678 Sera ID BIHI 4 1 LOT 7 1 Pos dr 1 25 5 Al AT Specificity Sample Date 12 212004 12 2 2004 Ad Wa 35 Patient Antibody Report Collected detailed reaction data and specificities for all samples linked with the specified Patient ID e Generated by selecting Report gt Patient Antibody Report from the main menu Figure A 2 Patient Antibody Report GRENFELD MAXIM Sample Date ABO MIC Tpos Patient Sera Report for GREN 2443 C at ID Lot Sera ID specificity Avi LS1PRA 008 P Overall m 6 500 400 5 000 LABScreen_V3 1_IVD Rev 1 pdf OLI UD 0176 022007 Appendix A LABScreen Reports iy 17 3 10 2 F neg Pos Tneg Hm F pos
38. allow to generate several of the most commonly produced LABScreen reports A number of other reports types are available and are discussed in Appendix A LABScreen Reports LABScreen V3 1 IVD Rev 1 pdf Chapter 2 Main Menu and Controls 11 of 72 OLI UD 0176 022007 3 7 07 Update Parameters Menu LABScreen program parameters can be set globally by selecting the appropriate Update Parameter functions accessed from the main menu These settings are discussed in Chapter 5 Parameters and Algorithms Many of these parameters can be temporarily set to local values on the sample level in forms accessible from the Analysis windows see Class I Analysis Results Window p 26 Main Window Controls The control panel icons and buttons allow you to move through records within a database table to perform basic file management operations and to access some of the most commonly used functions in the program The Navigation New record Cancel and Save icons are enabled when you select the Analysis Maintenance or Report menu options Many of these controls are conventional and require little comment Table 2 1 Main Menu Controls BL NN NEN NN Displays the first last record of the currently There are no keyboard equiva Li o active database table lents for this button Displays the previous next record of the cur The up down arrows have the EB rently active database table same effect Creates a new record in an editable table If the table is read o
39. ase Archiving Data The Archive Data option is available from the File menu You can archive Serum information Reaction information Class I or Class II analysis results From the Main menu select File gt Archive and Delete Data Select one of the data options listed above Although the windows for archiving each type of data differ in some respects the procedure for archiving and deleting records from these tables is basically the same for each type of data The examples in this chapter use the Serum Information screens Figure 4 1 Archive LABScreen Serum Information Dialog Box Archive LABScreen Serum Information i x Serum ID Beginning a1 wWa834 09 19 02 ending axe Include Deleted Records Delete Records after archive OK Cancel Specify the beginning and ending serum IDs of the range you want to archive or delete To archive a single record specify the same value for both the beginning and ending serum ID in the range Click the button to select specific serum IDs Specify whether to include records marked for deletion LABScreen V3 1 IVD Rev 1 pdf Chapter 4 File and Data Maintenance 39 of 72 OLI UD 0176 022007 3 7 07 6 Specify whether to premanently delete records marked for deletion after they have been archived 7 Supply a name for the archived file LABScreen automatically adds the arf extension Click OK to create the file A message confirms th
40. atio Count a z4 0 4 HH a Ed SoH b 16 6 3omt 6 15 8 zou b 15 5 SHE b 13 8 3c b 13 8 4c g e 56 b 16 6 CHE b 1lz 59 4 HH 67 of 72 Class I and Class Il Analysis Antibody Screening Reports Reversed Analysis report generated after reversing the NIH scores for the sample 8s become Is 6s become 2s etc The score rever sal or inversion in effect tests for the absence of antigens rather than their presence e Generated from within the Class I Class II Analysis Result window by clicking the button then selecting the Print option A detail of the report is shown below Figure A 14 Class and Class Il Antibody Screening Reports Reversed LABScreen Class I Antibody Screening Positive and Negative Reversed Patient s name HAPEUV SOREN Serum Id No Al Walse Patient Id HAPESE7S Face Percent positive Center Id GLENDALE Total valid reactions 55 Date zampled lz lz zu u 4 Number of pozitivez 5 Analysis gt Marmal J5siqnment Positive reactions are 56 9 Date tested De zz zund Overall strength index za 00 4 HLA Typing Control Values lt LS1PRR OOF gt NC 413 5 PC 7396 0 PC NC 17 9 Background De faalt Percent 8 l 73 Percent 5 6 q5 455 Percent 8 6 4 gt P4554 Percent 2 6 4 z 94 55 Awg Str Chi Fisher s specificity Score IP FP FN TN Incl Index Square z Tail 6 5 10 l5 d aa 43 40 au 5 11 031971 2s zb 00 63 051365 2s 00 51 08730 Z 0E
41. aw Data panel zr rr 200 Figure A 11 Sample Raw Data Report LABScreen Data CatlbD Lat LS1PRA 007 Session ID ABSCREEM DATI AFILESSESTPRA OU Cass Test Date 5 22 2004 10 06 14 AM Tested By Viv Nc Sample ID A1 Vv4136 PRA Percent 8 Percent 8 6 Beads ID Ren Reagent ID O01 MC 002 PC 003 0102 O04 C5014 OOS E6446 006 E2223 007 S001 opg S009 La Eq 76 O10 E15032 O11 E15254 O12 C 53003 L1 3 0099 hk hk h3 c CO M MDM E St c 059 O90 095 096 ogy 095 C5016 01 30 E2834 C4630 E12572 E185858 LABScreen V3 1 IVD Rev 1 pdf OLI UD 0176 022007 Rawdata 413 5 Pc 796 0 Pce Hc 17 8 54 5556 Or AF So 5 45 25 45 Ratio Count 413 5 1 0 FARR r 396 0 0 4 Bs S8 0 1 4 EI 366 0 0 5 JB H 3468 0 56 9 51 9254 0 13 8 45 tt 212 0 2 4 51 11690 24 0 4att 369 0 13 8 JAER 2462 0 44 oF aua 2 0 JAER 1055 0 2 3 FARR 6956 5 1 4 JAER Percent 5 6 4 Percent 5 6 4 2 Specificities A2 29 BS7 64 Bud B CWE 18 A2 24 B54 58 Eid B CVM 10 Ad 32 BBO 64 EG Cy ve 10 Ad 23 BB8 27 EMV4B CUM A3 33 BB 38 B46 CT 12 Aq 80 618 50 BWE CVO 5 Ad 69 635 49 Bud B CVF 12 A350 69 BH 73 EVE CVWA 5 17 A23 BS B37 72 Bud 6 C2 A3 B27 51 Bd CV 2 A2 29 B7 46 BYE CV 15 at RH 42 tH 1112 0 1 8 5534 5 5 3 arb OF 40 4226 0 a JI 395 5 1 1 jazz 10641 5 6 ar A431 B55 58 BW 4 5 Cv 4 Ald 34 B56 62 Ave CM 8 Ad 24854 67 Ave CVA 7 A3 bb BY et BV We CVYTS 1 A B46 6 Be CVA 7 As 36 B45 53 BV 6 Cv W
42. bodies are present in the tested serum LABScreen SA While the beads used in the other LABScreen tests may exhibit multiple specificities each bead in a LABScreen Single Antigen assay is specific for a single antibody Single Antigen assays are used to confirm the presence of specific antibodies suggested by an earlier PRA test About this Manual This manual contains information helpful in using the LABScreen software and consists of the following chapters Chapter 1 ntroduction the current document contains descriptions of the products in the LABScreen family an overview of the manual system requirements and installation instructions Chapter 2 Main Menu and Controls provides a survey of where to find various LABScreen analysis and maintenance functions Chapter 3 Analysis provides step by step instructions for performing both Class I and Class II analyses It includes details on changing computer assigned specificities adjusting cut off values and displaying reaction patterns Chapter 4 File and Data Maintenance explains how to perform such routine housekeeping tasks as archiving and retrieving data and packing and rebuilding the database Chapter 5 Parameters and Algorithms provides instructions for setting up global program parameters and includes the statistical formulas used in the LABScreen analysis Appendix A LABScreen Reports provides examples of LABScreen analysis results reports data reports and panel l
43. ckground value for test beads LABScreen V3 1 IVD Rev 1 pdf Chapter 5 Parameters and Algorithms 44 of 72 OLI UD 0176 022007 3 7 07 Table 5 4 Mixed Advanced Settings Control Legend States the initial default values LABScreen for reference for Class I Class Il and Other e g MICA Note that these are the initial and not the previous settings Class I Class Il Current cutoff values for Class I Class Il and Other Other e g MICA Figure 5 4 LABScreen Mixed Advanced Setting Dialog Box LABScreen Mixed Advanced Setting Ratio computation iz set az follows Ratio s n 8 n Sin Median for Bead zn Median for Bead 1 Brn Average Median for Bead zn for Negative minus Average Median for Bead 1 for Negative or minimum value specified default setting iz 50 Minimum Bn 50 00 Positive Ratio iz initialized to 1 5 and grey area ratio is setto 1 2 for both Class and Il Grey Area Positive Class 17 15 Class Il 17 15 Others 17 15 Cancel MFI to MESF Conversion Settings Table 5 5 MFI to MESF Conversion Settings Lot Use to locate database record containing con version curve slope and intercept Update legend Timestamp for selected record YYYYMMDDhhmm format MFI to MESF conver Activates conversion globally when selected Figure 3 1 Figure 3 4 sion activate MFI to MESF conversion panel Figure 3 3 is automatically opened during PRA SA and Mixed Data I
44. cpecificiliez Average TruePos FalzePos FalzeMeg TrueNeg R value Strength Chisquare Overall 7 155 22 0 2 31 0 5925 72 13 47 4 8 0010 M 0 168 1060 0 12 0 8 0010 EE 0 551 100 0 15 1 6 667 3 0 531 33 33 11 8 6 000 ao 0 5445 0 000 11 6 7 0010 31 0 791 50 00 23 2 Cat ID s Analyzed 007 Specificities Table this table Figure 3 14 displays statistical results for the antigens that qualify as specificities for the sample R Value gt 0 25 and Score gt 2 LABScreen displays its assignments in the Overall column LABScreen_V3 1_IVD Rev 1 pdf Chapter 3 Analysis 26 of 72 OLI UD 0176 022007 3 7 07 The area outlined in red contains editable fields You can delete a computer assignment or add your own antigens to check the effect of the change on the statistics If you attempt to include an antigen that does not belong to the panel LABScreen will display an error message When you make any edit to the table the small C Computer in the upper left changes to M Manual The product s used in the analysis are displayed in the Cat IDs Analyzed field beneath the table Percent Positives Panel this panel displays cumulative percentages for each category of NIH scores Figure 3 15 Note that the definitions of score assignment 2 4 6 8 reflect the global settings made in the Update Parameters Settings dialogs accessed from the main menu and do not reflect any local or temporary set tings made for the particular sample
45. cting Maintenance Serum Information from the main menu These latter tables contain data only for those samples whose analyses have been reviewed and Saved 1 e accepted from within a Class I or Class II Analysis Results window Figure 3 13 HLA Typing Information provides access to Class I and Class II serological typing results from LABScreen analyses and Class I and Class II DNA typing results from LABType analyses Each record is identified by Cell ID The Serological and DNA panels are editable forms in which you can modify the current typing results by making different Class I and Class II antibody and allele assignments A similar set of Cell Information tables can be accessed by selecting Maintenance HLA Typing Information from the main menu These latter tables contain data only for those samples whose analyses have been reviewed and Saved 1 e accepted from within a Class I or Class II Analysis Results window Figure 3 13 Analyze by Batch allows you to perform analyses on an entire previously imported PRA or Mixed Antigen batch or on selected sera contained in the database Serum selection criteria include By Class within the speci fied batch or By Range of contiguous serum records in the database Other search filters include by technician ID the ID entered when imported a LABScan data file test date catalog ID lot number or sera that have not yet been reviewed i e whose analysis results have not been accepted Sa
46. cting the Summary option in the Data Import Report dialog zr e200 Figure A 6 Mixed Data Summary Report CatlD Lot LSM12 012 Session ID 1 ABSCREEM DATAFILESSESMT2 012 OUTPUT Cay Tested By Vivi Test Date 05 26 2006 03 17 57 Default Background Values NC n n 5642 0 LABScreen_V3 1_IVD Rev 1 pdf OLI UD 0176 022007 Classi Classil MICA Rawdata Count 11 0 248 5 0 266 arl 256 1856 0 282 1 0 205 30 184 11 0 433 11 0 302 gel 234 r 240 Bb 252 1585 5 255 10 214 10 218 15 0 SBE Appendix A LABScreen Reports LABScreen Mixed Data Summary Rawdata Count 1082 0 243 0 9142 5 B26 5 10423 5 9537 5 56 0 1068 0 2170 9631 5 1U 354 5 5951 0 9965 0 darr 5 33 0 1070 0 60 of 72 127 111 1 att 1323 13x 1 att 1 att 191 159 133 13x 13x 13x 132 att Vii PCIHC Ratio 22 1 1142 8 71 5 55 4 9537 5 18 7 dr 1 18 7 100 3 1450 6 66 5 53 4 darr 5 33 0 71 3 PRA Data Report This report can be printed from the Analysis or Import windows When MEFS conversion is activated it contains MESF values 02702007 Figure A 7 PRA Raw Data Report LABScreen Data CatlDyLot LSIPRA D10 session ID YA ESTDATAXSTPRAITU CSS Test Date 04 24 2005 03 20 22 PM Tested By WAN Sample ID lt UNNAMED gt 9106 PRA MESF Beads ID Ren Reagent ID O01 O10 O11 O12 un 3 O14 O15 O16 on 7 O16 O19 DE O20 84 055 0S6 oa 055 059 O09 LABScreen_V3 1_IVD Rev
47. d which do not meet the R Value and Score LABScreen V3 1 IVD Rev 1 pdf Chapter 3 Analysis 27 of 72 OLI UD 0176 022007 3 7 07 criteria are displayed in the Chi Squares Table Figure 3 17 when you press the Chi Square button in the control panel Many samples display no remainder antigens Figure 3 17 Chi Square Table for Remainder Antigens speciticties d EE F M If you access the Chi Square table when the CREG display option has been invoked the Chi Square table displays the CREGs of the remainder antigens Figure 3 18 Figure 3 18 Chi Square Table for Remainder Antigen CREGs specificitiez TP PP F M e CREG Display some antigens shown in the Specificities Table may be member antigens in Cross Reactive groups Possible CREGs the complete listing of their known member antigens and abbrevi ated statistics are displayed at the lower right of the Analysis Results window More complete statis tics for possible CREGs can be displayed in the Specificities Table by toggling the CREG Agn buttons in the control panel Figure 3 19 CREG Display Possible CREG antibody s are Specificity TruePos FalseNeg R val 12 15 0 428 AT ASG AY AZS AZH R11 LABScreen_V3 1_IVD Rev 1 pdf Chapter 3 Analysis 28 of 72 OLI UD 0176 022007 3 7 07 Table 3 2 Analysis Window Controls ees IR Generates a Class or Class II Antibody Figure 3 16 Analysis Window v Print before save Screening Report when you Save the analysis Control
48. e 16 Appendix A LABScreen Reports 65 of 72 Class and Class Il Panel Listings Panel listings can be generated by selecting Report gt Panel Listing from the main menu then specifying the LABScreen product from a pick list You can preview the listing by right clicking on the print button or export the listing to an Excel file Figure A 12 Typical Antigen Panel Listing 3 ReagentID HLA DRE1 HLA DO HLA DRBS 4 5 Others NC Pi E177758 DR1 DR4 Pas nos DRS53 C4630 DR12 DRS Do4 Dos DR5Z istsgs DRO DRi Pas Doo DR53 C4613 DR13 DR15 Do5 DOs DR52 DRSL GOO87 DRS DRL Dae DOs DR5Z C4615 DR4 DR Doa DOs DR53 E5610 DR103 DRL Doz Dos DR5Z i069 DRS DR15 Das nos DR53 CS3722 DR4 DR14 Do Doa DR52 DE53 E6446 DR1 DR Doa DOs DRS53 E2538 DR11 DRL Dos DR5Z LABScreen V3 1 IVD Rev 1 pdf Appendix A LABScreen Reports 66 of 72 OLI UD 0176 022007 Class and Class Il Analysis Antibody Screening Reports Contains complete analysis data for the sample currently under analysis NIH scores NBG ratios and specificities for FPs ENs and TNs are also included in the report although only the TPs are shown in Figure A 13 e Generated from within the Class I Class II Analysis Result window by selecting the Print option Figure A 13 Class and Class Il Antibody Screening Reports LABScreen Class I Antibody Screening Patient s name Serum Id No Al W4ls6 Patient Id Pace Center Id Date zampled fof Anal ysis Computer Assigrment Date test
49. e count of the records archived Retrieving Archived Data The Retrieve Data option is also accessed from the File Menu It is effectively the same as the Archive Data function but only in reverse As such it requires no separate discussion Packing the Database Figure 4 2 Pack Database Options Pack Database E Pack option physically removes deleted records Select Information You wish to pack and then Click OK Click Cancel to Exit without performing this function M Serum Information Iw Analysis Results Readings Iv Cat Lot Information Antigens LABScan100 Rawdata Cell Information OK Cancel Use the Pack Database function to compress the tables shown in Figure 4 2 Note Using the Pack Database option permanently deletes all records that are marked for deletion Make sure that you want to remove these records before you use this option Rebuilding the Index If you experience circumstances that affect the integrity of your database tables you may be able to use the Index Rebuild function to restore your database tables This function is completely analogous to the Index Retrieval function and requires no separate comment LABScreen V3 1 IVD Rev 1 pdf Chapter 4 File and Data Maintenance 40 of 72 OLI UD 0176 022007 3 7 07 Chapter 5 Parameters and Algorithms Updating Global Parameters The Parameters Settings form Figure 5 1 is used to set global analysis parameters and report defaults It is accessed by
50. e print icon e Grey area results are indicated by three asterisks e Low PC NC ratios are indicated by double left angle brackets lt lt e Low bead counts are indicated by double pound signs LABScreen Reports Reports Menu Use these menu options to generate several of the most commonly produced LABScreen reports A number of other reports types are available and are discussed later in this appendix Patient Antibody Summary generates a summary Patient Sera Report see Figure A 1 Patient Antibody Summary Report for each patient specified by Patient ID A summary report lists just the positive Rxns and the specificities from all LAB Screen analyses associated with that patient The user can specify a single patient ID or a range of contiguous patient IDs Patient Antibody Report generates a detailed specificity report with overall statistics for the analysis Rxns of positive Rxns of strong Rxns etc and detailed data for each specificity TP FN FP and TN and statistics Inclusion Strength R Value and Chi Sq The user can specify a single patient ID or a range of contiguous patient IDs Patient HLA Report generates a summary report of HLA typings for all the cells associated with the specified patient s The user can specify a single patient ID or a range of contiguous patient IDs Panel Listing generates a panel listing for the specified product Cat ID and Lot CREG Listing generates
51. e user during anal ysis are presented You can revisit a Saved sample by selecting the appropriate single sample Analyze option and entering its Sample ID 3 Specify whether you are analyzing Class I Class II or both by checking the appropriate boxes e If you want to select specific Sample IDs within each batch check the Select Specific Samples check box to access the pick list shown in Figure 3 31 LABScreen V3 1 IVD Rev 1 pdf Chapter 3 Analysis 37 of 72 OLI UD 0176 022007 3 7 07 X EZ RO6213 x Am3b5 0l x W352 X meedo X TZ AEBS x BS 539 W1lod9 zx B X x m Done Select Mone Select ll Cancel e Select samples by selecting or clearing the X next to each sample ID Click Select All to include all samples in the range Click Select None to clear all check boxes 4 Click Analyze to proceed Note If your criteria for selecting multiple samples produces more than 96 matches the pick list in Figure 3 31 will appear even if you have not checked the Select Specific Samples option You can deselect samples to reduce the size of the group and thus reduce the processing time LABScreen V3 1 IVD Rev 1 pdf Chapter 3 Analysis 38 of 72 OLI UD 0176 022007 3 7 07 Chapter 4 File and Data Maintenance This chapter provides instructions for completing these tasks Archive and retrieve serum reaction and results information Export reaction and results information Pack the database Rebuild the datab
52. ed 06 22 2004 HLA Typing Control Values LSlIPRA OOF gt NC 413 5 POC 73s96 0 PC NCI17 9 Avg Specificity Score TP FP FH TH AL 6 5 7 y l 40 A74 7 0 Z 5 40 hrerall 6 6 3 5 l 40 Possible CHEC antibodyiz are Specificity TruepPos FalseNeq R val te lz 15 O 425 1LCZ gt 25 A A36 A9 AFG Az4 All True Positive CatID Lot Bead PBeagent AT LSIPRA 007 O08 CSO0S AT LSlPPRA OOF OSL El95109 AT LSlIPRA OOF 073 ELl 7230 LAT LSIPRA OOF 071 El8149 AT LSIPRA 007 OFZ 00TA AT LSIPRA 007 009 El77768 AT LSIPRA OOF O06 EZzzz27 472 LSlIPPRPA oo z 7 c lde 472 LSlIPRA 0007 073 El 7330 472 LSlIPRA 007 O80 El3 080 LABScreen V3 1 IVD Rev 1 pdf OLI UD 0176 022007 HLA A B C and Ewd Ew amp Percent positive Total valid reactions Mumber of positives Positive reactions are Overall strength index Backur ound De faalt Percent 8 Percent 8 6 Percent 8 6 4 Percent 2 6 4 z B SCE M Value Incl Index 55 0 558 a b zo 57 47 Oo 504 100 00 50 00 55 0 699 30 00 33 33 Els B50 Cie CWE Bile B49 B55 CW CWS Bild Bure B7z Bezol Cie CWO EWE Bl3 Bel Cie Bd Bre B47 B71 CWw1o Cul Bile B35 B49 CW CHlz2 Bild Bure BS Be CHI CUT EW4 EWE B44 BS C4 Chr BETA B7z Bezol Cie CHlO Bile Bs Bel Cle CLs Bile Appendix A LABScreen Reports F545 4 55 l4 bo zl 434 E 45 FG dbi 4 5553 oy 27 Chi Fisher s Square Tail 18 99 0 annis 11 94 O 013476 76 80 a 000z Pun P
53. emographic Information form this form can be accessed from several places within the program Clears all entries from Specificities Table Figure 3 14 Analysis Result including computer assignments Specificities Table The Eraser resets the entries in the Specific ities Table to the original computer assign ments Recalculates sample statistics after user edits to the Specificities Table LABScreen_V3 1_IVD Rev 1 pdf Chapter 3 Analysis 29 of 72 OLI UD 0176 022007 3 7 07 Table 3 2 Analysis Window Controls cont OO m C NEC RN Displays Revise Cutoff Raw Data grid only Figure 3 28 Color Coding in Revise Cutoff Raw Data Grid Displays Revise Cutoff Raw Data grid and Figure 3 27 and Figure 3 28 Reaction Histogram Note that local modifica tions to cutoffs are not indicated in Report footers and in the Percent Positives panel leg end Allows user to temporarily change the Reac Changing Parameters Locally Change Parameters tion Definition R Value and Score parame Temporarily p 36 E ters and report options for the Antibody Screening Report for the current sample Figure A 13 Global parameter changes must be made using the Update Parameters form from the main menu Displays the Reaction Listing by antigens Figure 3 21 Reaction Listing table by default only FPs and FNs are shown with CREGs TPs and TNs can optionally be displayed as well when the CREG option is invoked the Reaction Listing s
54. en meets the minimum exit the loop 4 Check the candidate antigens to see which has the highest value in the selected statistics In the case of a tie the first occurring antigen is ranked higher or highest 5 Add the antigen with the highest valuation to the list of selected specificities include associated statistics in the tabulation 6 Eliminate the selected antigen from the candidate list and remove all beads with that specificity from the data pool 7 Go back to Step 2 for the next iteration LABScreen V3 1 IVD Rev 1 pdf Chapter 5 Parameters and Algorithms 52 of 72 OLI UD 0176 022007 3 7 07 LABScreen_V3 1_IVD Rev 1 pdf OLI UD 0176 022007 Figure 5 6 Antigen Specificity Calculations Start Computation using available assessment pool Computed value x threshold Yes Select antigen with the highest computed value Assign selected antigen as specificity Eliminate beads from assessment pool with specificity already assigned Chapter 5 Parameters and Algorithms 53 of 72 3 7 07 This page intentionally left blank Appendix A LABScreen Reports LABScreen generates raw data analysis results and panel listing reports Many of the reports can be accessed from more than one point in the application Depending on layout requirements reports are generated either in portrait or landscape orientation When generating a report you can preview the report by right clicking on the appropriat
55. ent Advanced Settings PRA Parameter Settings The PRA Advanced Settings form Figure 5 2 is used to set cutoff levels and NIH score symbols for LAB Screen PRA analyses It is accessed by selecting Update Parameters gt LABScreen PRA Adv Settings from the Main menu Table 5 2 Update Parameters gt PRA Advanced Settings a GENE RN Legend States the formulas used to determine reac Reaction Positive Classification tion strengths in assigning NIH scores Guidelines and Formulas p 49 Cutoffs Cutoff constants used in the Reaction Classification formulas LABScreen V3 1 IVD Rev 1 pdf Chapter 5 Parameters and Algorithms 42 of 72 OLI UD 0176 022007 3 7 07 Table 5 2 Update Parameters gt PRA Advanced Settings i MEC RN Symbols NIH score symbols used the reports and data base tables By default these are 1 2 4 6 and 8 A bead is designated as Problematic is it does not have a reading These fields accept any characters in the extended ASCI character set Legend at bottom States the LABScreen default values for refer REN ence Figure 5 2 LABScreen PRA Advanced Setting Dialog Box LABScreen PRA Advanced Setting Positive classification computation iz set az folloyys negative Rxn to XA NCnHn aoc weak positive Rxn 244 NCOAOO FMC to amp E BCnh A OO eC postive Rxn B NCnA OO RNC to ACA OO FAC strong positive Rxn AC NEn OO PMC to amp D INCRIA OO ANC very strong positive
56. esces cued s 5 Product DOSet DUO us senex da erage weh Parme Ed SESS EO Rees fd od ohne UE Bene EEEN 5 LABS creen NIKCG 2o 23 depen d 5404 eara icd neri ces euesdansoeae paid S b Rad ERE 5 LABS a PRA TPR rnm 6 LAB O OA a E E T TITEL E E EE E eae ae ee 6 Abo t this Manal sse rasens eaaa Gre a ea prp E i ec R a Eaa cpu rapa bay B Aree es 6 System heGUINCINCING e cese rore soe teti a nR CELUM Se EER E IER Rc 6 Installing LABScreen Software 0 0 0 nee rns 7 Chapter 2 Main Menu and Controls LABSCICCH MORBUS asserere Reo urea s ER SER S RE ER dE ESSENSEN R 9 gir duin see seo reac r ew Brae ew teeny ce ere ae am oe Gaara ee epee goon ene hg es 9 lason EPPTP 9 AB Scan WOO NEO cauce sers ace ea ARX ESSPUREREESUGS ERES Oe nee eee see B EK 9 Analysis VIE D UE oa tcueaaeaeehediee coche ee we ee eh SAN GUS RASSE Cd ePi pA ens 10 Maintenance Menu PP 11 situ NIU e E S E E E S E EEEE ee eun 11 Update Parameters Men 4c ceochciacaaddoaduceedasivchadba ar Ed POR E De Rn 12 Mun Minn genus c MP v MM ATTMMTMET 12 Chapter 3 Analysis File Naming Conventions for LABScreen Products 0 cece eee eens 15 Importing PRA and Single Antigen Data 0 0 0 0 0 eens 15 Importing Mixed Antigen Data cc ehh 19 Importing Quantiplex MESF Beads Data For Investigational Use Only 21 Reprocessing a LABScan 100 Piles 224404062008
57. gative control serum The user would copy the sample results into the local background values then use the Save to function to save the NBG values to a BGV file j Sorts entries in Data Grid Reaction Histogram Reaction by descending NIH score sub sorts by descending bead number Sorts entries in Data Grid Reaction Histogram BeadlD by ascending bead number Sorts entries in Data Grid Reaction Histogram by descending raw data value You can adjust cutoff thresholds numerically or graphically Sorts entries in Data Grid Reaction Histogram NBG Ratio by descending NBG Ratio score Adjusting Cutoffs To adjust cutoffs numerically e Click the Cutoff button at the bottom of the Revise Cutoff Raw Data window to display the cutoff adjust ment entry fields Figure 3 26 Enter the desired values for each threshold The smallest increment the fields accept 1s O 1 e Click Apply to save your adjustments The new threshold values will be reflected in the Reaction Histo gram Figure 3 27 e Finally click Save in the tool bar at the bottom left to make the changes persistent The adjusted cutoff val ues apply only to the current sample Figure 3 26 Cutoff Adjustment Entry Fields PRA Weak Positive Strong Very Strong Positive Positive Positive curio 1 3 50 10 0 20 0 Apply Cancel Defaut To adjust cutoffs graphically e Grab a cutoff line and drag it to the desired location The coordinates in the tool
58. gen binding process can effect non specific binding e Patient therapies such as immunoglobulin IVIG Rituxan or thymoglobulin ATG Negative Control bead 001 median fluorescence should be lower than 1500 NC beads are blank not coated with HLA antigen NC bead fluorescence indicates the level of non specific binding Abnormally low NC values can be due to high albumin IgG ratios observed in dialysis and plasma pheresis patients Positive Control bead 002 median fluorescence should be higher than 500 and at least twice the NC value PC beads are coated with purified human IgG PC bead fluorescence indicates saturation of binding sites Low PC values may be due to washing techniques e Free IgG left in the assay can prevent the secondary antibody from binding to target e IgG PE signal can be diluted Low PC values may be due to dilution of IgG PE PRA Reaction Formulas NBG Ratio lt 1 5 NIH score of 1 negative NBG Ratio between 1 5 and 5 NIH score of 2 weak positive NBG Ratio between 5 and 10 NIH score of 4 positive NBG Ratio between 10 and 20 NIH score of 6 positive NBG Ratio above 20 NIH score of s strong positive Single Antigen Formulas of Working Range lt 15 NIH score of 1 negative of Working Range between 15 and 30 NIH score of 2 grey area of Working Range between 30 and 70 NIH score of 4 weak positive of Working Range above 70 NI
59. h at a later time you must reenter the manual values Importing Quantiplex MESF Beads Data For Investigational Use Only Note The new Quantiplex features are a Beta version only and may be subject to change when the actual OLI Bead product is released If you choose to include MFI to MESF data conversion you will need initially to create manually a separate lot for each Luminex LABScan analyzer from which you import LABScreen batch data Moreover you will need to update the values for that lot on the days when it is in use so you can associate a time stamped conver sion formula with the data collected on that day by that analyzer In order to create a MFI to MESF conversion lot carry out the following steps Locate a Quantiplex Bead Data sheet Figure 3 6 This is an Excel file that will be provided to you by the laboratory staff who run the LABScan 100 analyzers 2 Select Update Parameters gt LABScreen MFI to MESF Conversion Settings gt Update Initial Pairs to enable all the fields in the form LABScreen_V3 1_IVD Rev 1 pdf Chapter 3 Analysis 21 of 72 OLI UD 0176 022007 3 7 07 Figure 3 6 Quantiplex Beads Data File Quantiplex Beads Data Sheet Lot 1 12 8 2006 SFI Units if 7083 87 1036 1025 26855 sri 4932 49362 115350 31 21 67 22219 4560793 Use oU ul per test select regions 41 67 87 and 31 set sample volume 50 ul acquirelUU beads Use the Trimmed Mean fram Lurminex far Calculation
60. hows the CREGs of the member antigens present on each bead Displays any CREG with member antigens Figure 3 21 Reaction Listing among the detected specificities in the Speci with CREGs ficities Table Causes the Specificities Table to display detected antigen specificities default display Accesses a pick list containing all antigens Viewing Results by Antigen and alleles currently in the LABScreen data p 31 base upon selection of an antigen displays a Reaction Table listing all beads in the product that exhibit the antigen Excludes the specified antigens from analysis Excluding Antigens p 36 permanently or temporarily thus reducing 7 E the masking effect of those antigens on other antigens Excludes the patient s own antigens from the Exclude Own Antigens analysis if they have been entered in the m patient information LABScreen V3 1 IVD Rev 1 pdf Chapter 3 Analysis 30 of 72 OLI UD 0176 022007 3 7 07 True Positive F F F F F P Ren Ratio Count Cat B 21 2 136 B 17 4 107 B 14 4 123 Figure 3 20 Reaction Listing with CREGs Reaction Listing X v False Negative v False Positive True Megative List iare Bead HALA Typings LS1PRA OOF lt 011 gt 425 466 Bar Bf CVV Dd Eve LS1PRA OOF lt 031 gt 429 469 B38 B55 CVT OAT BWE LS1PRA OOF lt 014 gt 42 411 B13 BEZ GA CAG BV 4 Eve xFPz FP FM zFM eFM Fh FN zFM aFM Reaction Listing True Po
61. ilable in three principal versions which are listed below in the order of increasing i e finer resolution LABScreen Mixed The Mixed Antigen test contains a mix of both Class I and Class II antigens and is used to detect Class I and Class II antibodies in a single test The bead set in a Mixed Antigen panel consists of 10 to 17 beads 5 to 9 test beads with Class I antigens 3 to 5 test bead with Class II antigens plus negative and positive control beads Each of the test beads is coated with cloned antigens of up to eight different cell line The Mixed assay is typically used a preliminary screening test For reasons of economy the bead set is restricted in number If a Mixed analysis detects antibodies of interest the researcher will then subject the serum to PRA testing LABScreen V3 1 IVD Rev 1 pdf Chapter 1 Introduction 5 of 72 OLI UD 0176 022007 3 7 07 LABScreen PRA Panel Reactive Antibody tests detect antibodies and their specificities against the HLA antigens in panels of Class I or Class II antigens separately or Class I and Class II antigens simultaneously Class I and Class II PRA panels contain approximately 55 and 35 beads respectively not counting a positive and negative control bead in each lot In the PRA and Mixed assays the antigens on a single bead may react with multiple antibodies thus providing an ambiguous screening result by itself By comparing the reactions of multiple beads it is possible to determine which anti
62. iscussed above in Step 7 4 Accept the default background values or specify Negative Background values by entering a Specific Sample ID as discussed above in Step 4 5 Alternatively clear both check boxes and manually enter background values for the Control Beads and Test Beads Note that the numeric legends for the Test Beads to the left of the input fields change accord ing to the definition of the product Figure 3 5 but are updated only after you have pressed one of con LABScreen V3 1 IVD Rev 1 pdf Chapter 3 Analysis 20 of 72 OLI UD 0176 022007 3 7 07 trols at the bottom of the form Report Only Import Data etc Check these legends to make sure you are dealing with the right product The input fields are initially pre populated with the default values for the product Figure 3 5 Manually Entering Negative Background Values L Use Default Background values Negative Background Use Specific ID FC 5000 45 50 Controls ME 026 Jo Test Beads Un 45 022 50 ps eso 028 s55 rf 6 You can now proceed to the analysis See Analyzing Data p 24 Note If you enter any Negative Background value for a test bead that is less than the minimum value thresh old set globally in the LABScreen Mixed Advanced Settings LABScreen will automatically bump the value up to the threshold value By default this value is 50 Manually entered Negative Background values are not persistent Thus if you reprocess the same batc
63. istings System Requirements The following is required to successfully install and run LABScreen software e IBM compatible PC with Pentium III processor e 5 Gigabyte hard drive e 256 megabytes RAM e VGA display e Windows 2000 or Windows XP operating system e Luminex interface software version 1 7 or IS2 2 LABScreen_V3 1_IVD Rev 1 pdf Chapter 1 Introduction 6 of 72 OLI UD 0176 022007 3 7 07 Installing LABScreen Software If you do not yet have the LABScreen software installed on your computer follow these instructions to install it 1 2 3 A Close all other Windows applications before starting the installation process Insert the One Lambda CD in your CD ROM drive From the Windows Task Bar select Start gt Run From the Run window in the Open box type in the location of your CD ROM drive followed by a back slash and Labscrn31 exe e g D Labscrn31 exe e Or click the Browse button to select your drive and then double click the Labscrn31 exe file Click OK to begin the installation process The install wizard usually places the LABScreen software in your C labscreen directory LABScreen V3 1 IVD Rev 1 pdf Chapter 1 Introduction 7 of 72 OLI UD 0176 022007 3 7 07 This page intentionally left blank Chapter 2 Main Menu and Controls This section provides a brief overview of menu functions and links to detailed discussions in subsequent sec tions of the manual All program functions can be accessed from
64. l 3 0 8510 84 7 43 5 AA2B 8 000 18 33 0 343 100 0 6 5 A33 8 000 14 3 0 395 100 0 8 0 ABE 8 0010 10 33 0 468 100 0 10 3 A25 8 0040 ri 33 0 497 100 0 10 6 A34 7 0040 5 33 0 498 50 00 3 9 Ba 6 000 3 3 0 606 0 000 13 9 e If we consult the Chi Square display for the remainder antigens in this sample we can see a listing of all the other antigens that have at least 2 FPs and an R Value over 0 20 Figure 3 23 Figure 3 23 Chi Sq Values for A10 233 34 W912 Remainder Antigens speciticties TP FP F M e We can use the Reaction by Antigen function to see how the remainder antigens reacted in the test by click ing the By Agn button and choosing a remainder antigen from the Antigen Lookup pick list Figure 3 24 Antigen Lookup Antigen Lookup Select Cancel Mame e The Reaction Listing for the A11 antigen is shown in Figure 3 25 Since the point of this feature is to let us view all beads carrying the antigen all reaction types are displayed by default The complete specificities of all beads that also have the A11 specificity are listed LABScreen_V3 1_IVD Rev 1 pdf Chapter 3 Analysis 32 of 72 OLI UD 0176 022007 3 7 07 Figure 3 25 Reaction Listing for A11 in A10 233 34 W912 Reaction Listing for lt A11 gt X w True Positive v False Negative v False Positive JW True Negative List More Ren Ratiocount Cat Bead HALA Typings lt TR gt FR 304 143 LSIPRA 007 lt 021 817 433 B51 B54 CN CAMO Bid BYE FP B 14 4
65. lation Strength Strength the percent Strength 1s calculated by the formula LABScreen V3 1 IVD Rev 1 pdf Chapter 5 Parameters and Algorithms 50 of 72 OLI UD 0176 022007 3 7 07 Ng Eq 5 8 Str 100 Nrp Inclusion Index Inclusion Index the Inclusion Index is the ratio Nop Eq 5 9 Incl Nrp Nery Values range from 0 to 1 Average Average this is the weighted Average of all reactions that are classified as true positives If both 8s and 6s are considered positive the Average is calculated as 8N 6N6 Eq 5 10 Avg Ns N6 where N is the number of reactions in the true positive group If 4s are also considered positive both the numerator and the denominator contain corresponding terms for the 4s Ave SNg 6No 4N Eq 5 11 e C Net NaN Fisher s Two Tail Fisher s Two Tail Tail analysis refers to the analysis of the low frequency parts of a distribution for example the tails or ends of a distribution curve Tail analysis 1s an appropriate statistic tool to use when analyzing anti gen frequencies because of the large number of antigens and the small number of times any given antigen is detected during the assay The Fisher probability test is used to examine the significance of the association between two variables in a 2 x 2 contingency table The need for the Fisher test arises when we have data that are divided into two catego ries in two separate ways reactio
66. m ID field If you have entered an invalid Sample ID LABScreen will generate a warning message e Close the csv file making sure not to save it as an Excel file When you run an analysis of the sample whose values you have specified to use as background values note that all the results are zero This 1s expected inasmuch as all readings for that well are cancelled out by the background values you are now using for the entire batch 7 If you do not want to include MFI to MESF conversions in the reports skip to Step 8 In some cases you will want to use MESF calibration data for the Luminex machine that was used to col lect the data for the samples you are analyzing in order to compare samples collected on different dates To activate MFI to MESF conversion e Check the unlabeled box at the upper right of the Import Report dialog to display the MFI to MESF conversion table which shows the current values used in the regression routine that generates the con version formula e The slope and intercept of the fitted curve a straight line are shown above the table The RR value indicates the goodness of fit By default the calibration curve is based on readings for the five beads listed in the first column of the table Figure 3 3 e You can modify the values used to generate the conversion formula by selecting the Update Parame ters gt MFI to MESF Conversion Settings option from the main menu LABScreen V3 1 IVD Rev 1 pdf Chapter 3
67. mport Reprocess LABScreen V3 1 IVD Rev 1 pdf Chapter 5 Parameters and Algorithms 45 of 72 OLI UD 0176 022007 3 7 07 Table 5 5 MFI to MESF Conversion Settings Bead ID Data pairs table accommodates up to 7 beads Note that the Bead ID is not necessarily always in ascending order although the MFI values are Initially these fields are read only Current MFI MFI values used together with MESF values for generate the Slope and Intercept of the conversion formula Minimum RSQ Minimum threshold for goodness of fit The conversion formula is generated from the MFI MESF data paris using a least squares fit If a linear formula cannot be generated within this threshold the RSQ value in the Calc panel is flagged with asterisks Maximum MFI Data pairs of beads with MFI values above this limit are discarded before the equation fit is made Calc Reruns the least squares fit using the valid values in the cells then displays the updated MFI to MESF conversion formula Update Initial Pairs Enables Bead ID and MESF entry fields Save As Saves new conversion formula to database table after user enters name for database record up to 9 characters Restore from Default Reloads values from currently selected data base record LABScreen V3 1 IVD Rev 1 pdf Chapter 5 Parameters and Algorithms 46 of 72 OLI UD 0176 022007 3 7 07 Figure 5 5 MFI to MESF Conversion Settings LABScreen MFI to MESF Conversion settings oes MyLoto21 4
68. n gt Import Reprocess LABScreen SA Data Duplicates the function of Update Parameters gt Update Parameters 13 of 72 3 7 07 This page intentionally left blank Chapter 3 Analysis File Naming Conventions for LABScreen Products The first step in carrying out a LABScreen analysis is to import a batch file of serum information that has been generated by the LABScan 100 analyzer In some cases the LABScreen Catalog IDs and Lot numbers are not immediately obvious to the user The length of product names in LABScreen Software is limited to twelve characters To accommodate product files name for LABScreen Single Antigen products which may consist of as many as three products with dif ferent Catalog IDs and Lot numbers these file naming conventions are followed The Catalog IDs and Lot number combinations of two groups may include one of four alphanumeric charac ters The absence of an alphanumeric character indicates that the lot number is the same for both groups e A The lot as written for group one and the lot number as written plus one for group 2 e B Thelotas written for group one and the lot number as written plus two for group 2 e Z The lot as written for group one and the lot number as written minus one for group 2 e Y The lot as written for group one and the lot number as written minus two for group 2 Please refer to the following examples e LSIAI amp 2 007 designates L51A01 Lot 007 LS1A02 Lot 007 e LSI
69. n positive or negative antigen present or absent and the sample sizes are small or the data are very unequally distributed among the cells of the table e g Table 5 6 The question posed regards the probability of the distribution of the observed reactions presented in a 2 x 2 table such as shown in Table 5 6 The probability is calculated using the formula p ABI CI DC D Eq 5 12 NA B C D Note that 0 1 Values range from 0 to 1 with O indicating no probability of occurrence and 1 00 complete probability Antigen Specificity Calculation The antigen specificity table is generated as shown in Figure 5 6 LABScreen V3 1 IVD Rev 1 pdf Chapter 5 Parameters and Algorithms 51 of 72 OLI UD 0176 022007 3 7 07 Retrieve all data needed for the selected sample and product combination reactions associated raw and normalized data and the antigens associated with each bead compile a list of candidate antigens from which the specificities will be determined 2 Setup a2 x 2 table of TPs TNs FPs and FNs for each candidate antigen as shown above in Table 5 6 TP FP TN and FN Reaction Designations Compute appropriate statistics including Chi Square Fisher s Two Tail Score Strength and Inclusion Index for each antigen 3 Check the statistics against the specified threshold Currently the user may choose either the R Value or the score as the primary threshold criterion and the other as the secondary criterion If no antig
70. nly this icon will be disabled Discards all changes made since the last In many places in the program Save this button closes the current window and returns to the previ ous window when analyzing multiple samples it ends the current analysis and proceeds to the next Saves any changes to the database Generally using the Save icon to commit a change avoids the Save confirmation dialog box The Alt S keystroke combina tion is equivalent to using an OK or Save button Exits the program when in the home page Elsewhere in the program this button serves only to close the currently active window Cell Info Update Duplicates the function of Maintenance HLA Typing Information Sera info Sera Info Update Duplicates the function of sspe Maintenance Serum Information E Analysis Duplicates the function of Analysis gt Analyze Class and Class Il Cell Info LABScreen V3 1 IVD Rev 1 pdf Chapter 2 Main Menu and Controls 12 of 72 OLI UD 0176 022007 3 7 07 Table 2 1 Main Menu Controls BL NENNEN To N Import PRA Import PRA Import Mixed Import SA Import S4 Update Parameters LABScreen_V3 1_IVD Rev 1 pdf Chapter 2 Main Menu and Controls OLI UD 0176 022007 Duplicates the import function of LABScan gt Import Reprocess LABScreen PRA Data Duplicates the import function of LABScan gt Import Reprocess LABScreen Mixed 10 17 bead format Data Duplicates the import function of LABSca
71. ons that are common to all Windows applications LABScan 100 Menu Use these options to import data files generated by the LABScan 100 analyzer or to reprocess the data in files that have already been imported Typically the main purpose of reprocessing is to analyze imported data using a different product lot From the user s perspective the main distinction between data importation and data reprocessing is as follows e when importing data the user locates and selects a Luminex csv file by means of a standard Windows find file dialog e when reprocessing data it is only necessary to select the session name of the file from a drop down menu that contains session names of all the csv files currently loaded into the database Conveniently the default session names of the imported files are the same as their original filenames In both cases you must supply the appropriate catalog ID s and lot number s for the assay In all other respects import and reprocessing steps are identical These options are available LABScreen V3 1 IVD Rev 1 pdf Chapter 2 Main Menu and Controls 9 of 72 OLI UD 0176 022007 3 7 07 Import Reprocess LABScreen PRA SA Data import reprocess csv files for full PRA SA assays These may be either Class I Class IL or combined Class I and Class II assays Import Reprocess LABScreen Mixed 10 17 bead format Data import reprocess csv files for 10 17 Bead Mixed assays Mixed assays always contain a mix of
72. oring does not include a 6 reaction classification These fields accept any characters in the extended ASCI character set Legend The legend at the bottom of the form states the LABScreen default values for reference Figure 5 3 LABScreen SA Advanced Settings Form LABScreen SA Advanced Setting B Signal ta background ratio for bead n Yn Mn hMz1 Mi h 9n hn Median for testing serum on bead n M1 Median for testing serum on bead 1 N21 Median for Neg Control serum an bead 1 Mon Median for Meg Control serum on bead n Cut off for Yn for a testing serum is defined as 4 of working range defined between highest Yn and lowest Yn Yn cutoff value is ASC highest Yn lowest sms lowest Yr Problematic Medative Grey Weak Strong Positive Positive 30 0 PALAI cutoffs A 0 00 Symbols T 4 15 0 AB AL Positive Positive Cancel Cutotts were initialized to 15 30 70 Symbols were set to be 0 1 2 4 8 respectively 4 B Positive Mixed Advanced Parameter Settings The Mixed Advanced Settings form Figure 5 4 is used to set Grey Area and Positive Reaction criteria for Mixed analyses It is accessed by selecting Update Parameters gt LABScreen Mixed Adv Settings from the Main menu Table 5 4 Mixed Advanced Settings NEL NEL NNNM NN Legend State how Background values are defined Reaction Positive Classification Guidelines and Formulas p 49 Minimum Sets minimum default ba
73. ort it will be in the A 4 format To restore the report size back to the default letter size launch the Letter size report batch file LABScreen V3 1 IVD Rev 1 pdf Appendix A LABScreen Reports 70 of 72 OLI UD 0176 022007 Index Symbols low bead count 55 grey area results 55 lt lt low PC NC ratio 55 A Antigen exclusion temporary vs permanent 36 Antigen Specificity Calculation algorithm flowchart 51 ATG anti thymocyte globulin 49 Average defined 51 B Bead count minimum 49 C Catalog ID name length limitations 15 Chi Sq 55 formula 50 Table 27 CREG Display 28 CREG Selection UNOS and or Fuller 41 D Database packing 40 Date Format changing 42 Disclaimer 5 E Epitope analysis 11 F File extensions arf 40 Fisher s Two Tail formula for 51 LABScreen V3 1 IVD Rev 1 pdf OLI UD 0176 022007 G Global Parameters modifying 41 l J IgG PE 49 Inclusion Index 55 defined 51 Index rebuilding 40 Installation Instructions 7 L LABScan 100 reprocessing 24 LABScreen Menus synopis of functions 9 LABScreen Mixed adding new lots 11 enabling 4 bead analysis 10 LABType allele assignments 11 Main Menu file 9 reports 55 MESF Molecules of Equivalent Soluble Fluorophores 10 MFI Median Fluorescent Intensity 18 N NC values causes of low 49 Negative Background from ID 18 P Packing the database 40 Patient HLA Re
74. port generating 70 PC values causes of low 49 Product Description LABScreen Mixed 5 Index LABScreen PRA 5 LABScreen SA 5 Public Antigens 11 Q Quantiplex Beads Data 22 R Range Rule 10 Reaction Definition 30 Rebuilding the Index 40 Remainder Antigens Chi Sq table 27 CREGs 28 Report paper size changing between letter and A 4 70 Reports Panel Listing 70 Patient Antibody Report 69 Reprocessing Data 24 Rituxan 49 R Value 55 defined 50 S Saved Accepted sample results where to find 11 Score defined 27 50 Single Antigen Working Range defined 48 Strength 55 defined 50 System Requirements 6 T Tail analysis 51 Thymoglobulin anti thymocyte globulin 49 TPs FPs TNs FNs 2x2 Table 50 71 of 72 This page intentionally left blank
75. ppendix A LABScreen Reports 61 of 72 SA Raw Data Report This report can be printed from the Analysis or import windows When MEFS conversion is activated it contains MESF values Figure A 8 Single Antigen Raw Data Report 02 15 2007 LABScreen Data CatlD Lot LE1A182 001 Session ID JL ABSCREEN DATAFILESSESTAT 2 CSw Test Date 08 30 2002 03 12 05 PM Tested By AAN Hc 845 Pc 45871 Pe Ne 54 4 Sample ID P1 08 30 02 w Positive Percent 8 5 00 Percent 5 4 28 576 Percent 8 47 43 7575 Ratio Maximum Ratio 202 30 Minimum Ratio 1 69 Cutoff far 8 0 142 173 Cutoff far 4 B1 88 Cutoff for 2 laos MESF MESF Beads Lott DRAFT10 Processed on 200702151558 Y 24175 21 8 20 305198x Beads ID Ren Reagent ID Ravwdata Ratio Count Specificities O01 HC 84 5 5692 0 1 0 562 002 PC 4597 0 97519 45 40 Sra O03 FR ADTUT T362 5 153673 2 i at on4 FR ADZUT gSa OF 1 455 4 28 5 246 Dos R A0206 14006 288570 7 128 5 O06 R ADU 964 0 23750 4 6 9 DO RAT101 993 0 24338 3 78 O08 FR A2301 3158 1 6300 0 m z z 4 1 1 1 OFF ura ura Lat Le ies Dies uad RAF S01 118 0 5582 5 H3 BB RE13512 113 0 i 5470 7 l BG BE RB1513 2r 0 9800 5 l BrT Bd RAO 111 0 6430 1 Hia Be B2101 123 0 6673 5 Bet BWE FR EBBZLT 131 0 6836 2 B201 BiG R AZ S37 5 150980 3 F ASSN 600 0 163539 3 h h ee k k k LABScreen_V3 1_IVD Rev 1 pdf Appendix A LABScreen Reports 62 of 72 OLI UD 0176 022007
76. s uses only that one By default both types are used Specifies which report elements are included Figure A 13 Class and Class ll by default in the Antibody Screening Report Antibody Screening Reports the preset default is TP FN and FP only Figure A 15 Class and Class ll The 2 X 2 option generates a separate report Antibody Screening Reports that lists the statistics for all antigens in the 2 x 2 Listing assay in the order of the highest R Value to the lowest Headings Institution specific information that is included in report headers LABScreen V3 1 IVD Rev 1 pdf Chapter 5 Parameters and Algorithms 41 of 72 OLI UD 0176 022007 3 7 07 Figure 5 1 Global Parameter Settings Form Parameter Settings Negative Below or Positive Above Equal to LAB Screen Reaction Definition E E Problematic Negative Weak Positive Positive Strong Positive very Strong Positive are indicated by symbols 0 1 2 4 5 8 respectively opec Selection F val ar Score Initial R secondar select Above R value 0 25 acnre F CREG Selection CREG Type Used UNOS Report List I True Pos I FalseNeg Panel Listing 2x2 M False Pos True Neg Panel Listing by Ratio Headings One Lambda Inc 21001 Kittridge St Canoga Park cA 81304 818 702 0042 Save Cancel Changing the Date Format The Select Data Format function accessed from the Update Parameters menu is self explanatory and requires no comm
77. selecting Update Parameters gt Update Parameter from the Main menu Reaction Definitions Specificity Selection Methods and Report Header Information Table 5 1 Update Parameters gt Parameter Settings Comes o tmm o RR Reaction Definition opecifies NIH score threshold for determining positive reactions in Class and Class II analyses Displays currently defined symbols for differ ent strengths of reactions in PRA and SA which are set in the Advanced Settings forms Specificity Selection Determines whether R Value or Score is to be H Value p 50 Score p 50 used to make the first specificity assignment and then subsequent assignments The software calculates R Values for all pos sible antigens then selects the antibody spec ificity with the highest R Value Once the first specificity has been select by the R Value method the Score calculation is used to assign the rest of the specificities Be advised that it may be admissible to use the R Value calculation for both Primary and Secondary assignments but it is not recom mended to use the Score method for the Pri mary assignment If you choose to change the threshold values you should perform validation testing based on the frequencies in your population CREG Selection Determines whether the analysis will use Figure 3 13 Class Analysis UNOS or FULLER CREGs If no entry is Results Window made the analysis uses both If one or the other is entered the analysi
78. sitive Rn Bele eye eyo co List Figure 3 21 v False Negative More Ratio Count Cat ID 22 3 123 aa 0 4 0 6 a 6 9 6 3 6 2 Sb 43 46 a Sr 40 S1 42 3b LS1PRA LS1PRA LS1PRA LS1PRA LS1PRA LS1PRA LS1PRA LS1PRA LS1PRA Reaction Listing with CREGs Bead HALA Typings OOF 27 120 163 220 28C SC INTL1 INTL OOF lt 030 gt 103 260 IMTL3 007 n25 120 102 103 28C 202 5C 7C BB INTL2 INTL3 OOF lt 021 gt 101 152 103 210 220 28C 5C 8C BB IMTEZ INTL3 007 098 gt 120 101 172 1073 5C BB INTL INTL3 007 n22 120 101 102 103 210 280 202 SC INTE2 INTE3 007 005 120 101 102 1073 27C TC ac BB INTL2 INTL3 OOF naue 101 1652 103 210 220 28C SC BB INTLS OOF lt 020 gt 120 101 102 1073 27C 286 202 7C BB INTL INTL3 Viewing Results by Antigen This option allows you to inspect the behavior of antigens that do not meet the R Value and Score criteria Consider the analysis results of the sample shown in Figure 3 22 e In this example the six antigens that meet the R Value and Score criteria are shown in the Specificities Table see Antigen Specificity Calculation p 51 LABScreen V3 1 IVD Rev 1 pdf OLI UD 0176 022007 Chapter 3 Analysis 31 of 72 3 7 07 Figure 3 22 Class Analysis Result for A10 233 34 W9121 Class I Antibody Analysis Result for 410 33 34 W9171 i cpecificitiez Average TruePos FalsePoz FalzeMea TrueMedg R value Strength C hisSmqguare Overall T b854 19 3 i
79. tip are Bead Position NBG Ratio Figure 3 27 The revised values also appear in the Cutoff Adjustment Entry Fields Figure 3 26 e Click the Save File icon at the bottom left to preserve the changes LABScreen_V3 1_IVD Rev 1 pdf Chapter 3 Analysis 34 of 72 OLI UD 0176 022007 3 7 07 Color Coding in Revise Cutoff Raw Data Grid When you adjust cutoff values beads in the data grid that have undergone a change in NIH score are marked with a red X and their new NIH scores are also shown in red See bead 17 in Figure 3 28 LABScreen applies the yellow alert color to any NBG ratio that is 0 25 of the cutoff between NIH scores of l and 2 Figure 3 27 Reaction Histogram with Cutoff Lines 140 00 120 00 100 00 80 00 Very strong Positive Cutoff MOL EDGE Ul us EESSEGMEINGSINNIGSLIASINGEGILLELLQELIIELUL mewn C SEL LIC GLO C GIL CIL AGE I Pek gg Strong Positive Cutoff 40 00 4 16 63 60 m Positive Cutoff 20 00 L m m ommo mmo mm mmo mmo mmo umo mo mmo m mmm mom I ae TI 004 amp 2 24 B54 56 Dg 6 C 10 13 ostive Cutoff 0 00 H l l 1 002 Pas 001 Neg 52 Bv 6 CVE B 012 amp 3 B27 51 EV CWA 25 032 430 31 E13 71 B46 CWE 9 036 amp 32 36 653 61 B46 CVI2 4 021 411 33 B51 54 By 5 CVA 10 086 411 B38 75 HAWG CT 84 081 amp 1 23 649 55 Bd 6 CW 9 009 41 69 635 49 Bd B CF 12 022 411 23 849 52 Bd CVV 125 069 225 68 852 62
80. ts Table for each sample in turn Figure 3 13 Class I Analysis Results Window You can move to the next serum by clicking the Save button the Return button or by pressing Return Analyze non prompt mode to printer imports all serum test data performs analyses and sends reports for each serum directly to the printer without user intervention thus printing reports for all imported samples automatically Note that although the serum analyses have been performed by the program the results have not yet been Saved accepted by a researcher Use this option when you want LABScreen to carry out preliminary analysis of a batch of samples that you will review at a later me 5 You can interrupt and terminate the analysis cycle by pressing S between samples LABScreen V3 1 IVD Rev 1 pdf Chapter 3 Analysis 16 of 72 OLI UD 0176 022007 3 7 07 Table 3 1 Test Figure Table LABScreen PRA Data Import Report Luminex Data File Tech ID JALABSCREENM_DATAFILESILSTPRA_OO CSW Cat ID 1 Cat ID 2 LS2PRA OOF one C Print Antigen Distribution Table with Report L Analyze non prompt mode to printer Previews Report First Q Analyze after Import L Use Default Background Values Negative Background from ID A25 i587 5 Report Only Import Data Import amp Report Exit Figure 3 1 Import PRA SA Data LABScreen PRA Data Import Report Luminex Data File Tech ID JILAB SCREEN _DATAFI
81. ved Note When specifying multiple sera your selection must observe the Range Rule the end of range selec tion must follow the start of range selection in the Serum listing To specify a single Patient enter the LABScreen V3 1 IVD Rev 1 pdf Chapter 2 Main Menu and Controls 10 of 72 OLI UD 0176 022007 3 7 07 same Patient ID in both the start and start of range fields The Range Rule applies throughout the selection functions of LABScreen Analyze Class Class Il Class amp Il use this to perform the respective analyses on a single serum Analyze Single Antigen allows you to perform a batch analysis on a previously imported Single Antigen data file or on selected sera within the database Serum selection criteria are 1dentical to those described above for PRA or Mixed Antigen Analyze by Batch option Analyze Epitopes not yet implemented Maintenance Menu This menu provide tools for reviewing and editing a variety of records in the LABScreen database Options include Readings NIH scores catalog lot data and raw data for each tested serum in the database The serum ID cat alog lot data test date and reader ID can also be edited here Antigen names loci pertinent leukocyte cell types for the antigen or allele T or B and allelic subtype infor mation for each antigen and allele in the database Public Antigen member antigens in CREGs T or B and edit fields for modifying or adding member anti

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