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lipoprint® system user's manual

1. Krauss RM Low density Lipoprotein Subclass and Risk of Coronary Heart Disease Curr Opin Lipidol 1991 2 248 252 Muniz N Measurement of Plasma Lipoproteins By Electrophoresis On Polyacrylamide Gel Clin Chem 1977 23 1826 1833 Third Report of The National Cholesterol Education Program NCEP Expert Panel on Detection Evaluation And Treatment Of High Blood Cholesterol In Adults Adult Treatment Panel III Executive Summary NIH Publication No 01 3670 May 2001 Stein E Greer IA Myers GL National Cholesterol Education Program Recommendations for Triglyceride Measurement Clin Chem 1995 41 1421 1426 Sattar N Greer IA Louden J Lindsay G McConnel M Shepherd J Packard C Lipoprotein Subfraction Changes In Normal Pregnancy J Clin Endocrinology Metab 1997 82 2483 2491 2 29 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM C h rs pte r 3 SAMPLE ANALYSIS TOPICS SEE PAGE Scanning 3 1 Image Analysis Automated 3 7 Report Generation 3 13 Image Analysis Manual 3 21 Printing 3 26 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4gc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS SCANNING Preparation The scanner tray holds the Lipoprint tube template which accommodates up to 44 gel tubes for scanning The gel tubes
2. i T HoT a naene DAVII i T user s manual clinical research lipoprint system user s manual clinical research The Lipoprint LDL System with the Lipoware Clinical analysis software has been cleared by the FDA for the analysis of lipoprotein subfractions in human serum or plasma Upon customer request for supporting ongoing research selected units of the Lipoprint LDL System have been modified with additional software for Research Use Only RUO The RUO software is not intended for diagnostic purposes as it has not been reviewed by the FDA The following symbols are used in labeling for Quantimetrix Corporation Lor wd IVD amp Co Re A El European Conformity Lot Number Manufactured by For in vitro diagnostic use Biological Risk Contents of kit Catalog No See Product Insert Authorized Representative Store at Use by last day of month Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM Table of Contents CHAPTER 1 CHAPTER 3 ABOUT THIS SYSTEM SYSTEM OPERATION What To Do If You Are A New User 1 1 Scanning 3 1 Abbreviations 1 2 Image Analysis Automated 3 7 Technical Updates 1 3 Report Generation 3 13 Product Information Technical Support 1 3 Image Analysis Manual 3 21 Components 1 4 Printing 3 26 Principles Of Operation 1 5 Assembly 1 6 Login 1 11 CHAPTER 4 TROUBLESHOOTING CHAPT
3. bar then click Print A printer window will appear Click Print Note txt files will not print as a spreadsheet if dragged to the printer icon 3 26 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM C h a pte r 4 TROUBLESHOOTING TOPICS SEE PAGE Introduction 4 1 Electrophoresis Process 4 2 Scanning 4 3 Analysis 4 4 Report Generation 4 5 Printing 4 6 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM TROUBLESHOOTING INTRODUCTION The following chapter is included in the Lipoprint User s Manual in order to help the user solve problems that may arise while using the system 4 1 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM TROUBLESHOOTING ELECTROPHORESIS PROCESS PROBLEM POSSIBLE CAUSE SOLUTION Loading gel polymerization incomplete Time of polymerization Follow product insert Hemolyzed sample Do not use sample Diluted loading gel Blot buffer completely Polymerization inhibitors in sample see product insert Bad sample do not use Tube not touching light Repeat test Protruding gel air bubbles Shipping damage m
4. 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS IMAGE ANALYSIS Manual Analysis Cont 6 Enter the position number of the tube to be analyzed and click OK NAAN a position number for tube 1 7 Enter the sample ID of the specified tube and click OK NAAN Sample ID for tube 1 Bill Shakespeare Cancel OK 3 22 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS IMAGE ANALYSIS Manual Analysis Cont 8 Enter the TC values for the specified sample and click OK 100 mg dL is the default cholesterol value fr O ry Tot Chol for tube 5 5 Bill 258 00 9 To analyze the sample click OK on the dialog box Place the cross cursor slightly above the VLDL band and click once Move the cursor slightly below the HDL band and click once as shown in fig 3 1 click center of tube slightly above VLDL then slightly below HOL WLDL CHD HDL Figure 3 1 3 23 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS IMAGE ANALYSIS Manual Analysis Cont 10 Improper band selection dialog box will appear by double clicking above VLDL Sel
5. 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS PRINTING Lipoprint reports can be printed in two ways one report or multiple reports Start by opening the images 1 folder on the desktop to locate the Lipoprint report files to be printed To print one Lipoprint report at a time double click specific report file icon to open report Choose File on 2 upper menu bar highlight and click Print A printer window will appear Click Print Printer HP Photosmare 5300 sees Presets Standard ot Copies zz M Collated Pages AII Ofrom 1 toi 1 Paper Size U5 temer 77 8 50 by 11 00 inches Orientation ta 8 Center image EE O Scale 100 EQ n Gis Scale To Fit Print Entire Image O Fill Entire Paper images per page 1 C Print 1 copies per page Cancel C Print ory Supplies 3 Printing can also be performed by dragging the selected Lipoprint report to the Printer icon at the dock _ Pm 1 4 ae ETEA CO la 4 To print multiple reports select the reports to be printed Marquis or A la carte highlight the desired Lipoprint reports see pg 3 13 step 2 then drag them onto the Printer icon at the dock Note Do not print more than eight reports at one time To print a txt file double click on the file and it will open as a spreadsheet Choose File on the upper menu 5
6. 104 0 LDL 2 17 49 33 37 112 1 LDL 3 14 28 21 25 119 0 LDL 4 14 5 10 12 120 0 LDL 5 12 6 5 83 3 LDL 6 9 4 2 50 0 LDL 7 15 7 2 28 8 HDL 29 35 32 35 109 4 Table 13 Observed vs expected values for blended sample AC alc AC mg dL mg dL rae ao recovery VLDL 35 13 24 25 104 2 MiC 37 16 27 25 92 6 MidB 35 11 23 18 78 3 mica 15 16 16 14 875 tort 21 54 38 36 94 8 LDL 2 17 38 28 34 121 4 LDL 3 14 T 11 16 145 4 LDL 4 14 7 11 157 1 tors 12 6 5 83 3 LDL6 9 4 2 50 0 WDL7 15 C 7 2 286 mL 29 40 35 37 105 7 2 25 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT SPECIFIC PERFORMANCE CHARACTERISTICS Sensitivity Sensitivity of the Lipoprint System was defined as the minimum concentration of HDL C and total LDL C that can be detected reliably It was determined as the intersection of the lower 95 confidence interval of the mean with the x axis when plotting a dilution series of expected vs actual values for the individual lipoprotein parameters VLDL sensitivity is gt 2 02 mg dL HDL sensitivity is gt 3 65mg dL and Total LDL sensitivity is 8 30 mg dL Interferences Potentially interfering substances were spiked into serum
7. tube 8 Quantimetrix LIPOPRINT SYSTEH a M Flawed Report 6 Sub fraction cholesterol 0 magfdl LS Range Range LDL Profile not indicative of Type A presence of small dense LDL Total Chol mgidl 100 2200 LDL Chol mgidl 0 lt 130 Reference ranges derived from 125 serum samples that met the NCEP ATPIII fuidelines for desirable lipid status LDL lt is comprised of the sum of cholesterol in Mid bands C through A as well as all the subfractions Lipoprint Profile with red background Solution Quit Lipoware and re launch to reanalyze tube template 3 20 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS IMAGE ANALYSIS Manual Analysis To reanalyze specific gel tube s follow steps through 3 on pages 3 7 to 3 8 4 After selecting the template image to be analyzed the user will be prompted for the mode of operation For manual analysis enter m and click OK NOM Automated a or manual m analysis Cancel OK 5 Enter only the number of tubes that need to be manually analyzed then click OK Note The tubes need not to be adjacent to each other e g if the total number of tubes is 3 Tube 1 tube 17 and tube 19 or any other combination of tubes can be analyzed 3 21 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA
8. 0 91 7 00 10 0 81 8 10 2 22 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT SPECIFIC PERFORMANCE CHARACTERISTICS Inter Assay Precision Cont Table 8 Inter assay precision data for LDL subfractions LDL 1 SD CV LDL 2 SD CV LDL 3 SD CV LDL 4 SD CV LDL 5 SD CV LDL 6 SD CV LDL 7 SD CV Sample N Mean Mean Mean Mean Mean Mean Mean mg dL mg dL mg dL mg dL mg dL mg dL mg dL 1 80 41 1 61 3 92 11 11 47 13 50 N A N A NA N A N A 2 80 37 1 43 3 86 39 15 3 85 14 2 69 19 21 N A NA N A N A 3 80 29 1 1 3 79 24 11 73 7 21 20 1 33 6 65 20 1 21 6 05 N A N A N A 4 80 24 0 88 3 67 22 11 48 6 73 17 0 95 5 59 20 0 69 345 24 0 62 2 58 17 2 05 12 06 4 1 39 33 90 Linearity Linearity studies were conducted with two serum samples fortified with either HDL C or LDL C isolated by ultracentrifugation Sample 1 LDL C 695 mg dL HDL C 260 mg dL VLDL C 140mg dL Sample 2 LDL C 163 mg dL HDL C 178 mg dL VLDL C 38mg dL 2 23 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6
9. 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT SPECIFIC PERFORMANCE CHARACTERISTICS Linearity Cont Serial dilutions were prepared using a physiological human serum albumin solution The concentrations observed for total LDL C HDL C and VLDL C were compared to the expected values Table 9 11 Percent recovery was calculated as follows recovery observed value expected value x 100 Table 9 Dilution linearity for LDL Dilution Observed LDL Expected LDL Recovery mg dL mg dL 0 695 695 100 0 10 627 626 100 2 30 466 486 95 9 50 351 348 100 9 70 195 209 93 3 95 59 69 85 6 Dilution Observed LDL Expected LDL Recovery mg dL mg dL 0 163 163 100 0 5 155 157 98 7 10 151 148 102 0 50 84 89 94 4 90 15 15 100 0 95 13 13 100 0 Table 10 Dilution linearity for HDL Dilution Observed HDL Expected HDL Recovery mg dL mg dL 0 260 260 100 0 10 218 234 93 2 30 174 182 95 6 50 117 130 90 0 70 73 78 93 6 90 25 26 96 2 Dilution Observed HDL Expected HDL Recovery mg dL mg dL 0 178 178 100 0 5 170 169 100 6 10 162 160 101 3 50 95 89 106 7 95 11 9 122 2 97 5 5 3 94 3 Table 11 Dilution linearity for VLDL Dilution Observed VLDL Expected VLDL Recovery mg dL mg dL 0 140 140 100 0
10. 10 129 126 102 4 30 99 98 101 0 50 75 70 107 1 70 48 42 115 5 90 22 14 157 1 Dilution Observed VLDL Expected VLDL Recovery mg dL mg dL 0 38 38 100 0 5 36 37 97 3 10 34 35 97 1 50 24 24 100 0 90 17 13 130 8 95 13 11 4 114 0 2 24 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT SPECIFIC PERFORMANCE CHARACTERISTICS Dose Response Serum sample A containing a wide range of subfractions was blended with an equal volume of sample B containing primarily LDL 2 to give AB The samples were analyzed with the Lipoprint System the cholesterol concentrations of the individual fractions were determined and the observed values were compared with the expected values Table 12 Similarly a serum sample A containing a wide range of subfractions was blended with an equal volume of serum sample C containing primarily LDL 1 to give AC The samples were analyzed with the Lipoprint System and the cholesterol concentrations of the individual fractions were determined and compared with the expected values Table 13 Table 12 Observed vs expected values for blended sample AB A B AB mg dL mg dL al pee recovery VLDL 35 20 28 29 103 6 Mid C 37 20 29 28 97 6 Mid B 35 15 25 21 84 0 Mid A 15 15 15 15 100 0 LDL 1 21 29 25 26
11. Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT SPECIFIC PERFORMANCE CHARACTERISTICS Accuracy by Correlation Cont A similar comparision was performed between Ultracentrifugation 8 Quantification and the Lipoprint LDL System A population of 40 serum samples with LDL C ranging from 66 211 mg dL Lipoprint and 68 218 mg dL B Quantification HDL C ranging from 29 91 mg dL Lipoprint and 28 90 mg dL 8 Quantification as well as VLD C ranging from 9 5 49 mg dL Lipoprint and 6 57 mg dL 8 Quantification was evaluated Tables 17 19 Table 17 Lipoprint LDL B Quant LDL N 40 40 mean mg dL 130 8 130 0 SD mg dL 30 14 30 42 regression Lipoprint LDL 0 933 LDL8 Quant 9 430 2 0 887 Table 18 Lipoprint HDL B Quant HDL N 40 40 mean mg dL 53 5 53 5 SD mg dL 15 29 15 71 regression _ Lipoprint HDL 0 944 HDL quant 3 030 2 0 941 Table 19 Lipoprint VLDL B Quant VLDL N 40 40 mean mg dL 24 7 22 9 SD mg dL 10 34 12 61 regression Lipoprint VLDL 0 689 VLDLg quant 7 990 2 0 8216 2 28 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT REFERENCES 1 10 11 12
12. Reminder dialog box will disappear To obtain the best scan quality out of ScanWizard Pro please calibrate your scanner Click the Calibrate button now to exit SOS apan you will be to recalibrate your scanner to obtan Consistent scanning results specity how often you wish to be v Every 7 days Do not click Calibrate 3 3 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS SCANNING Operation Cont 3 A window will come up that requires the user to enter a file name next to Save As for each template of gels to be scanned The scanned images template should be saved in the Lipoprint Data folder on the desktop To locate the Lipoprint Data folder click on Desktop on the left hand column of the Save scanned image as window Select Lipoprint Data folder on the center column To start scanning click on Save at the bottom right side of the window Caution Confirm that scans are always saved in TIFF format NAA Save scanned image as Save As File name a faie 2m 1 Lipoprint Data z Q search A WW Desktop P 7 Images P 3 Documents ro 3 Lipoprint Data gt Dy macintosh HD h D Library p Music b z d Pictures t E Desktop FF aamin J Mopicatons 5 documents rane i Pictures 1 t jae Format TIFF ve Auto number file nam
13. Total Cholesterol must be entered manually Sample ID must not exceed 30 characters Click OK to continue AAO j Sample ID for tube 1 Prof Albert Einstein 3 10 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS IMAGE ANALYSIS Automated Analysis Cont 8 Enter the Total Cholesterol for the specified sample and click OK Note The Total Cholesterol of a given sample must be obtained externally e g clinical analyzer Default value is set to 100 mg dL perenne O N 9 After all the sample ID s and cholesterol values have been entered the program will proceed with the analysis and create a Lipoprint Report The Lipoprint reports will be automatically saved in the images folder on the desktop Note If an error occurs while entering the Patient ID or TC Cancel the entry process While cancelling the entry process a sequence of dialog boxes will appear Follow instructions in the dialog box click Don t Save and close all windows that follow Start over at step 2 pg 3 8 3 11 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS IMAGE ANALYSIS Automated Analysis Cont 10 After the analysis has been successfully comple
14. Warnick GR Wood PD National Cholesterol Education Program Recommendations For Measurement of High Density Lipoprotein Cholesterol Executive Summary Clin Chem 1995 41 10 1427 1433 Bachorik PS Ross JW National Cholesterol Education Program Recommendations for Measurement of Low Density Lipoprotein Cholesterol Executive Summary Clin Chem 1995 41 10 1414 1420 Griffin BA Caslake MJ Yip B Tait GW Packard CJ Shepherd J Rapid Isolation Of Low Density Lipoprotein LDL Subfractions From Plasma By Density Gradient Ultracentrifugation Atherosclerosis 1990 83 1 59 67 Otvos JD Measurement of Lipoprotein Subclass Profiles by Nuclear Magnetic Resonance Spectroscopy In Handbook Of Lipoprotein Testing Rifai N Warnick GR Dominiczak MH Eds AACC Press 1999 2nd Edition Washington DC Pages 609 623 Austin MA Hokanson JE Brunzell JD Characterization Of Low Density Lipoprotein Subclasses Methodologic Approaches And Clinical Relevance Current Opinion In Lipidology 1994 5 6 395 403 Rajman I Kendall MJ Cramb R Holder RL Salih M Gammage MD Investigation of Low Density Lipoprotein Subfractions as A Coronary Risk Factor In Normotriglyceridaemic Men Atherosclerosis 1996 125 2 231 242 McNamara JR Campos H Ordovas JM Peterson J Wilson PW Schaefer EJ Effect of Gender Age and Lipid Status on Low Density Lipoprotein Subfraction Distribution Results from the Framingham Offspring Study Arteriosclerosis 1987 7 483 490
15. dL mg dL mg dL mg dL mg dL mg dL mg dL 1 12 36 0 60 1 67 10 1 68 16 80 N A N A N A N A N A 2 12 28 0 91 3 25 32 0 70 2 19 14 1 65 11 79 N A N A N A N A 3 12 21 0 51 2 43 19 0 88 4 63 18 0 38 2 11 19 0 86 453 N A N A N A 4 12 24 0 86 3 58 22 0 80 3 64 17 0 28 1 65 20 0 49 2 45 24 0 4 1 72 16 0 72 4 62 4 0 68 17 89 Inter Assay Precision Four samples were tested in duplicate twice a day for 5 days in 4 electrophoresis chambers using a single lot of Gel Tubes The corresponding precision results for inter assay are shown in Tables 6 8 Table 6 Inter assay precision data for HDL LDL VLDL Sample N HDL C SD cv LDL C sD cv VLDLC sp cv Mean mg dL Mean mg dL Mean mg dL 1 80 60 149 2 49 94 141 1 50 10 0 90 9 40 2 80 46 145 3 15 137 1 73 1 26 T 0 91 827 3 80 32 1 52 4 75 160 2 03 1 27 33 2 35 7 12 4 80 50 2 32 4 69 178 2 79 1 57 23 1 97 857 Table 7 Inter assay precision data for Midband subfractions Sample N Mid C SD CV Mid B SD CV Mid A SD CV Mean mg dL Mean mg dL Mean mg dL 1 80 4 1 23 8 79 10 0 87 8 27 19 2 00 10 90 2 80 18 1 99 11 06 13 0 86 6 62 16 133 831 3 80 2 2 84 13 63 22 1 04 4 73 17 1 27 747 4 80 28 187 7 79 13
16. mg dL High Risk LDL C includes all the lipoprotein particles with d gt 1 006 to 1 063 kg L such as VLDL remnants IDL Lp a and LDL 2 These particles correspond to the sum of Midbands and the LDL subfractions resolved by the Lipoprint System Subfraction Cholesterol Expected normal values for the individual subfractions on the Lipoprint System were established as follows Self declared healthy individuals N 273 aged 18 to 85 years 166 females and 107 males 47 Caucasian 18 Hispanic 16 Asian 5 Black and 14 undeclared were recruited These volunteers were required to fast for 12 hours Whole blood was collected into a serum tube via venipuncture Serum samples were then used to test for indicated lipid parameters and to generate the lipoprotein profiles with the Lipoprint LDL system Exclusion criteria included diabetics individuals taking lipid lowering medication and those with recent heart attacks Additionally pregnant women were excluded from the study due to their changed lipid status 12 Only samples that met the NCEP guidelines ATP III for desirable lipid status 10 namely TC lt 200 mg dL LDL lt 130 mg dL HDL gt 40 mg dL and Triglycerides lt 150 mg dL 11 were used to determine the expected values The normal samples N 114 aged 18 to 84 years 32 male and 68 female were used to establish expected normal values defined as the 95 confidence interval mean 2 SD for each lipid parameter as obtained on th
17. recommend reading the following sections e Components on page 1 4 gt Provides an overview of system reagents hardware and software e Principles Of Operation on page 1 5 gt Describes the electrophoresis process scanning and analysis e Lipoprint LDL Kit on pages 2 1 to 2 29 gt Provides detailed instructions for setting up and using the gel tubes gt Provides storage recommendations for reagents and buffer solution e Sample Analysis on pages 3 1 to 3 26 gt Provides detailed steps involved in using Lipoware 1 1 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM ABOUT THIS SYSTEM ABBREVIATIONS A Ampere CAD Coronary Artery Disease CD ROM Compact Disk Read Only Memory CHD Coronary Heart Disease FAQ Frequently Asked Questions HD Hard Drive HDL High Density Lipoprotein HDL C High Density Lipoprotein Cholesterol IDL Intermediate Density Lipoprotein IDL C Intermediate Density Lipoprotein Cholesterol LDL Low Density Lipoprotein LDL C Low Density Lipoprotein Cholesterol mA Milliampere MHz Million Cycles Per Second Megahertz ML Milliliter pL Microliter OTS Off The Shelf PDF Acrobat File Format SCSI Small Computer System Interface TC Total Cholesterol TG Triglyceride USB Universal Serial Bus V Volts VLDL Very Low Density Lipoprotein VLDL C Very Low Density Lipoprotein Cholesterol 1
18. samples at the concentrations listed in Table 14 and tested next with uncompromised samples 4 replicates Hemoglobin at concentrations above 200 mg dL and heparin at typical concentrations found in heparinized plasma collection tubes were found to interfere with the Lipoprint test Table 14 Interferences Interferant Conc VLDL C Recovery LDL C Recovery HDL C Recovery mg dL mg dL mg dL mg dL none 17 1 1 100 115 1 9 100 42 2 4 100 bilirubin 20 15 0 9 89 115 2 2 100 45 1 6 107 hemoglobin 500 28 0 3 165 106 2 0 92 40 2 0 95 hemoglobin 200 18 0 2 106 116 0 7 101 40 0 6 95 niacin 2 5 16 0 7 94 115 1 5 100 43 2 0 102 EDTA 200 16 1 0 94 114 0 4 99 44 1 2 105 Heparin 14 U ml 20 0 4 118 113 1 4 98 41 1 9 98 Serum vs Plasma Comparison Serum and EDTA plasma samples from 37 patients were compared on the Lipoprint System The cholesterol values mg dL of all lipoprotein fractions and subfractions N 322 up to 12 per sample generated by the Lipoprint System were plotted and correlated Cholesterol 0 995 Cholesterol serum 1 158 r 0 971 plasma 2 26 QuantimetrixX 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT SPECIFIC PERFORMANCE CHARACTERISTICS Sample Stability Serum samples N 22 coveri
19. 2 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM ABOUT THIS SYSTEM TECHNICAL UPDATES Technical updates inform our customers of current information product improvements new products related to the operation of the Lipoprint System and related laboratory techniques Technical updates will be produced periodically and mailed to you as they become available Store the updates in the section labeled Technical Updates PRODUCT INFORMATION TECHNICAL SUPPORT To Reach Us By Telephone 7 00 am 4 30 pm Pacific Time Monday Friday 800 624 8380 1 310 536 0006 By Fax 800 845 1834 1 310 643 0180 On the Web www lipoprint com We strongly encourage you to visit our web site to learn more about our products E mail lipoprint 4qc com 1 3 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM ABOUT THIS SYSTEM COMPONENTS Reagents e Lipoprint LDL Kit 100 Tests e Liposure Lipoprotein Control Hardware e Digital Scanner e iMac Computer e Color Printer e Preparation Rack e Preparation Light e Electrophoresis Chamber e Power Supply 120V 220V e Rimming Tool e USB Printer Cable e Firewire Cable e Scanner Template Software e ScanWizard Pro e Lipoware e AppleWorks 1 4 Quantimetri
20. 21 References 2 29 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 e www 4gc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT INTENDED USE AND INDICATIONS FOR USE The Quantimetrix Lipoprint System LDL Subfractions Kit Lipoprint LDL Kit is a device intended to measure lipoprotein cholesterol for lipoprotein fractions and subfractions from VLDL to HDL in fasting serum or plasma with a Total Cholesterol concentration of gt 100 mg dL Lipoprotein cholesterol measurements are used as an aid in evaluating lipid metabolism disorders when used in conjunction with other lipid tests patient risk assessment and clinical evaluation 2 1 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT SUMMARY AND EXPLANATION OF THE TEST Plasma lipoproteins are spherical particles responsible for the transport of cholesterol triglycerides and phospholipids There are five major lipoprotein classes Chylomicrons very low density lipoprotein VLDL intermediate density lipoprotein IDL low density lipoprotein LDL and high density lipoprotein HDL Low HDL cholesterol is a strong independent predictor of coronary heart disease CHD 1 Increased LDL cholesterol LDL C has been identified as a major risk factor for CHD 2 It is known that the lipoprotei
21. 2A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT SUMMARY AND EXPLANATION OF THE TEST In a study of 109 patients with myocardial infarction MI Austin et al 5 showed that LDL subfraction Pattern B was associated with a threefold increased risk of MI independent of sex age and relative weight Krauss 8 reported similar findings Rajman et al 6 also reports on the risk factor associated with low density lipoprotein subfractions in normotriglyceridemic men 2 3 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT TEST PRINCIPLE The Lipoprint LDL Kit consists of e Precast linear polyacrylamide gel stacking gel and separating gel in a glass tube Fig 2 e Liquid loading gel with a lipophilic dye e Buffer salts Loading Gel Stacking Gel VLDL Cc B Midband A 1 7 Large LDL 3 J Small LDL HDL lt End of Gel Separating Gel N OU Figure 2 Lipoprint gel tube schematic 2 4 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT TEST PRINCIPLE The dye binds proportionally to the relative amount of cholesterol in each lipoprotein 9 The prestained lipoproteins subsequently undergo electrophoresis During the first phase of the e
22. DL TC mg dL mg dL mg dL mg dL mg dL mg dL mg dL mg dL mg dL mg dL mg dL mg dL range 5 69 9 40 5 37 5 34 9 77 7 55 0 35 0 28 0 11 58 215 26 137 104 319 mean 24 4 23 2 14 6 18 6 46 5 31 5 9 2 4 7 5 5 134 8 54 2 219 4 SD 13 50 5 65 5 21 6 14 15 13 10 98 7 50 6 05 3 6 27 17 18 05 35 04 95 0 12 0 4 2 6 4 16 2 9 5 0 0 0 80 5 18 2 148 6 range 51 34 7 25 0 30 9 76 8 53 5 24 2 16 8 16 189 1 90 2 290 2 N 141 141 141 141 141 141 115 44 8 141 141 141 Number of samples showing the respective fractions 2 20 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT SPECIFIC PERFORMANCE CHARACTERISTICS Precision Four samples were tested for intra assay and inter assay variability respectively Samples with low medium and high HDL C and LDL C were selected Sample 1 Low LDL C high HDL C and homogeneous LDL pattern LDL 1 and 2 only Sample 2 Medium LDL C medium HDL C and slightly dispersed LDL pattern LDL 1 2 3 Sample 3 High LDL low HDL and dispersed LDL pattern LDL 1 2 3 and 4 Sample 4 High LDL intermediate HDL C and dispersed LDL pattern LDL 1 through 7 Intra Assay Precision Samples were tested in replicates of 12 the maximum capacity of the electrophoresis chamber Precision results
23. E Category None C Auto Corrections g D Range Automatic E WEB Points No Correction W Tone Curve No Correction BE BAC NoCorrection W Color No Correction Filters Automatic Color Restoration Default 78ppi 100 ICROTEK 022 St Sick t create a new job or to toggle scan job selection Use Control Click to Mi temporarily override scan frame lock settings ScanWizard Pro Start Screen Note A Settings window will appear at the left side of the ScanWizard Pro window DO NOT CHANGE any of the Settings in this window Changes in the Settings will make the scanned image unusable for analysis 3 2 QuantimetriX 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS SCANNING Operation Cont Note A Calibration Reminder dialog box may appear as shown below Please calibrate your scanner s To obtain the best scan quality out of ScanWizard Pro please calibrate your scanner Click the Calibrate button now to exit ScanWizard Pro and run the Microtek Scanner Profiler to start calibration im the future you will be prompted to recalibrate scanner to obtain Consistent scanning results Please specify how yOu wish to be If the Calibration Reminder dialog box appears go to the drop down arrows select Never and click on Skip After clicking on Skip the Calibration
24. E tube 12 Quantimetrix LIPOPRINT SYSTEM f HDL Chyomicron Re eet SO dade ag 15 Sub fraction ert lien 12 mgtdl HI HI HI HI Lo Bae Be a so amon o aange Renga s22 23 5 35 5730 Range LDL Profile not indicative of Type A presence of small dense LDL Total Chol mg dl 100 lt 200 LDL Chol mgfdl 63 lt 130 Reference ranges derived from 125 serum samples that met the NCEP ATPIII puidelines for desirable lipid status LDL C is comprised of the sum of cholesterol in Mid bands C through A as well as all the subfractions Lipoprint Profile showing chylomicron Chylomicron presence of bubbles or artifact above VLDL Solution Perform manual analysis as indicated in pages 3 21 to 3 24 Be sure to click center of tube slightly above VLDL excluding chylomicrons bubbles or any other artifact 3 17 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS REPORT GENERATION Flawed Report 4 SAMPLE tube 1 Quantimetrix LIPOPRINT SY STEM ii Sub fraction cholesterol mgidl Al Ml At Al L Reference Lan Fe Reference Range s2 a 1 ST a 6 Oo OO Range LDL Profile not indicative of Type A presence of small dense LDL Total Chol mg dl 100 lt 200 LDL Chol mgidl 86 lt 130 Reference ranges derived from 125 serum samples that met the NCEP ATPIII puide
25. ER 2 Introduction 4 1 LIPOPRINT LDL KIT Electrophoresis Process 4 2 Scanning 4 3 Intended Use And Indications For Use 2 1 Analysis 4 4 Summary And Explanation Of The Test 2 2 Report Generation 4 5 Test Principle 2 4 Printing and Trash 4 6 Product Description 2 6 Warnings And Precautions 2 8 Specimens And Specimen Collection 2 8 RESEARCH USE ONLY Assay Procedure 2 9 APPLICATIONS Quality Control 2 13 Qualitative Results 2 14 Quantitative Results 2 16 PARTS AND ACCESSORIES Limitations 2 18 Expected Values 2 19 Specific Performance Characteristics 2 21 ONE YEAR WARRANTY References 2 29 Quantimetrix 2005 Manhattan Beach Bivd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM C h a pte r 1 ABOUT THIS SYSTEM TOPICS SEE PAGE What To Do If You Are A New User 1 1 Abbreviations 1 2 Technical Updates 1 3 Product Information Technical Support 1 3 Components 1 4 Principles of Operation 1 5 Assembly 1 6 Login 1 11 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4gc com MO92002A 6 10 LIPOPRINT SYSTEM ABOUT THIS SYSTEM WHAT TO DO IF YOU ARE A NEW USER This manual is written with the assumption that you know how to operate an iMac computer If you are not familiar with this computer refer to the Macintosh documentation included in this manual for specific operating information For an overview we
26. L Automatic Pipettor 4 Magnetic Stirrer 5 Parafilm 6 Graduated Cylinders Reconstitution of Reagents The electrolyte buffer solution is reconstituted by dissolving one vial of Lipoprint LDL Kit Buffer salts in 1200mL of distilled or deionized water Storages and Stability Gel tubes loading gel and buffer salts should be stored at 2 8 C Do not freeze With proper storage the reagents opened or unopened are stable until the date of expiration 2 7 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT WARNINGS AND PRECAUTIONS For In Vitro Diagnostic Use Only e Use the Lipoprint LDL Kit only in accordance with the Lipoprint Insert instructions e The loading gel solution contains acrylamide which is toxic when in contact with skin or swallowed Avoid inhalation and prolonged exposure to the loading gel solution e The loading gel is light sensitive and is packaged in an amber glass bottle e Avoid pipetting by mouth and any physical contact with the reagents or specimens e All samples reagents and controls should be treated as potentially infectious if ingested or absorbed through prolonged skin contact Precautions as they apply to your facility should be used for handling and disposal of materials at all times SPECIMENS AND SPECIMEN COLLECTION e Only fasting 12 hours samples should
27. Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT PRODUCT DESCRIPTION The Lipoprint LDL Kit consists of precast high resolution polyacrylamide gel tubes a loading gel solution containing a lipophilic dye and buffer salts Reagents and Materials Provided See Quantimetrix Catalog No 48 7002 A 100 test kit consists of 1 Lipoprint LDL Gel Tubes 100 tubes Polyacrylamide Buffer Preservative 2 Lipoprint LDL Loading Gel 24 mL Acrylamide N N methylenebisacrylamide Lipophilic dye Catalyst Stabilizer Buffer 3 Lipoprint LDL Buffer Salts 6 vials Tris hydroxymethyl aminomethane Boric Acid 4 Lipoprint LDL Product Insert 1 each 2 6 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT PRODUCT DESCRIPTION Lipoprint System Not Provided See Quantimetrix Catalog No 48 9150 1 Computer includes Lipoware Analysis Program Color Printer Digital Scanner Electrophoresis Chamber Power Supply 120V 220V Preparation Rack Preparation Light Oe DA ee Rimming Tool Liposure Lipoprotein Control Not Provided See Quantimetrix Catalog No 48 7060 Level 1 Material Required Not Provided 1 Distilled or Deionized Water 2 25uL Automatic Pipettor 3 200v
28. RINT LDL KIT ASSAY PROCEDURE 5 Apply 200uL of Lipoprint Loading Gel to each tube 6 Place a strip of Parafilm between the Gel Tubes and Preparation Rack Cover to avoid contamination Mix the Loading Gel with the specimen by inverting the Preparation Rack several times Fig 5 Figure 5 Mixing of loading gel and specimens 7 Set the Preparation Light upright with the bulb at the top Place the loaded Preparation Rack so the Loading Gel is touching the bulb Fig 6 Photopolymerize the Loading Gels for 30 minutes but no longer than 40 minutes Figure 6 Photopolymerization 2 10 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT ASSAY PROCEDURE 8 After the photopolymerization is completed remove each Gel Tube from the Preparation Rack and carefully insert it into the silicone adapters of the upper chamber While holding the Gel Tube by the side push it up until the loading gel end of the gel tube is flush with upper side of adapter Wetting the top of the Gel Tube makes insertion easier Avoid touching either end of the tube during this step If running less than a full chamber plug empty adapters with small glass tubes provided for this purpose Push tubes from top until flush with lower side of adapter Fig 7 Figure 7 Loading tubes 9 Place 1000mL of electrolyte buffer
29. SIS IMAGE ANALYSIS There are two modes for analysis Automated a and Manual m In Automated mode the software will identify the VLDL and HDL peaks without user interaction In Manual mode the user will be prompted to identify the VLDL and HDL bands on a digital image of the gel tube Automated Analysis 4 After selecting the template image to be analyzed the user will be prompted for the mode of operation Automated a is the default setting To proceed analysis in Automated mode select OK nnn Automated a or manual m analysis a Cancel OK 5 Enter the total number of tubes on the template and confirm with OK 24 3 9 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS IMAGE ANALYSIS Automated Analysis Cont 6 Prompt asks if the Sample IDs and Total Cholesterol TC values of all tubes will be entered manually Y is the default setting If Y is selected click OK If you do not want to enter Sample ID and TC values enter N click OK and the analysis will begin immediately program will name the tubes in sequence tube 1 tube 2 etc and use a default value of 100 mg dL for total cholesterol AAN Manually enter samples names TC Yy Cancel OK 7 If Y was selected then the Sample ID and
30. T SYSTEM _ i Sub fraction cholesterol mgidl fi At fi Ai LDL Profile not indicative of Type 4 presence of small dense LDL Total Chol tngidl 100 lt 200 LDL Chol mgidl 72 z 130 Reference ranges derived from 125 serum samples that met the NCEP ATPIII puidelines for desirable lipid status LDLC is comprised of the sum of cholesterol in Mid bands C through A as well as all the subfractions e Area not reported e Particle Size A for each LDL subfraction not reported e Mean LDL Particle Size not reported LDL profile is defined as indicative of type A or not indicative of type A 3 14 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS REPORT GENERATION Flawed Report 1 at 1212006 SAMPLE tube 1 Quantimetrix LIPOPRINT SYSTEM Sell 8 8 5 2 15 Sub fraction cholesterol mgtdl HI HI HI HI HI Lo Rino G2 a s Oe ooro o Benge Range s22 23 15 57 wooo a Range LDL Profile not indicative of Type A presence of small dense LDL Total Chol mg dl 100 lt 200 LDL Chol mgtdl 72 lt 130 Reference ranges derived from 125 serum samples that met the NCEP ATPIII puidelines for desirable lipid status LDL C is comprised of the sum of cholesterol in Mid bands C through A as well as all the subfractions Lipoprint Profile shifted to the right Ch
31. anner for analysis Allow the gel tubes to rest for at least 30 minutes but no longer than 2 hours before scanning Procedural Note Distortion of the separating gel surface and the VLDL band may occur during electrophoresis The distortion is corrected after the gels are removed from the electrophoresis chamber by carefully introducing the rimming tool from the top of the loading gel alongside the glass In a circular motion the surface of the separating gel The gel will return to its normal shape Avoid dislodging the loading gel when withdrawing the tool should the gel become dislodged fill the void with deionized distilled water 2 12 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT QUALITY CONTROL Reliability of test results should be routinely monitored with control materials that reasonably emulate performance on patient specimens Use of a quality control material is recommended with each run of patient samples Control materials are intended only as monitors of accuracy and precision The recovery of control values within the appropriate ranges should be the criterion in validating assay performance Quality control should be performed in conformance with local state and or federal regulations or accreditation requirements An appropriate quality control material Liposure is available from Quantimet
32. are loaded onto the template in the following sequence 1 Remove the template holder tray from the scanner bay 2 Insert the tube template with the label towards the handle and return tray to the scanner bay for gel tube loading 3 Carefully clean the gel tubes Place gel tubes onto the tube template slots beginning at position 1 and continue without leaving any empty slots between tubes Orient the tubes with the loading gel towards the number on the template 4 After all samples have been loaded carefully push the tray into the scanner bay Ensure that the tray is pushed until it stops Operation 1 Scanning is initiated by clicking on the ScanWizard Pro icon located at the dock 3 1 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS SCANNING Operation Cont 2 ScanWizard Pro start screen will appear To begin scanning click the Scan button Settings 000 ScanWizard Pro 4800 AF Scanner Overview Image File Scanner View Preferences Correction Help Job Positive A 7 m a e eee Positive ay Overview _ Prescan Scan 3 Q SC twee Coysa W ea P Eaa J Resolution 150 Flen E 3 w H E p Frame 8 000 x 10 00 inch fi pi Output 8 000 x 10 000 Scaling 100 0 18M8 Transform Fo aE M Sampling None
33. be used e Serum or EDTA plasma may be used See p 2 26 e Do not use heparin as an anticoagulant See p 2 18 e Samples can be kept for up to 7 days at 2 8 C e Freezing of the sample is not recommended However if a sample needs to be frozen it should be frozen cryogenically 70 C or below 2 8 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT ASSAY PROCEDURE 1 Prepare the electrolyte buffer solution as described by dissolving one vial of the buffer salts in 1200mL of deionized distilled water 2 Remove the Gel Tubes from the jar wipe off and place them in the Preparation Rack with the unfilled end up Fig 4 Avoid touching the ends of the Gel Tube or exerting any pressure on the gels since this will cause air bubbles to be introduced into the gel Do not use the Gel Tube if air bubbles appear inside or gel protrudes 3 Remove the storage buffer completely from the top of the gels by shaking the rack while inverted If necessary blot the end of the tube while the tubes are inverted in order to remove excess buffer from inside the tube 4 Apply 25yL of sample to each tube Fig 4 Figure 4 Sample application 2 9 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOP
34. cking on the file in the dock will position it back on the desktop screen tube 1 TED G amp D Ga G oP Previous Next Zoom Move Select Slideshow Sidebar 2 3 4 5 1 7 Sample ID Bar 3 Examine Lipoprint reports to verify proper analyses were attained see pg 3 14 to 3 21 A proper Lipoprint report will show small tic marks at the base of the graph corresponding to the apex of the VLDL and HDL peaks Reanalyze any unacceptable Lipoprint reports according to manual analysis steps outlined beginning on page 3 22 Reanalyzing a sample with the same patient ID will overwrite the previous report Note After analysis has been completed all files in the images folder should be transferred to the Lipoprint Data folder or to any other folder of your choosing The images folder should be completely empty of files before starting a new analysis To delete files select them and drag them to the Trash icon located at the dock Files can be recovered from the Trash by dragging them out unless they are permanently deleted by selecting Finder on the upper menu bar and then clicking on Empty Trash 3 13 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS REPORT GENERATION Lipoprint Profile Lipoware Clinical Version 81 1412006 SAMPLE ube 12 Quantimetrix LIPOPRIN
35. dondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 www 4gc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS SCANNING Operation Cont 6 To insure that a good image has been obtained check scanned template by opening the Lipoprint Data folder located on the desktop Double click the scan icon for the template to be viewed The template image can be Zoomed In or Zoomed Out by clicking on the zoom icon The template window may be enlarged by pulling the window outward by the right hand lower corner and by dragging the window outward with the mouse on the right hand lower corner A good gel tube template scan will show gel tube images with no artifacts e g bubbles smudges smears dirt dried buffer salts fingerprints in the area between VLDL and HDL Bubbles within the loading gel may be ignored because they are not within the analysis parameters therefore will not be graphed Bubbles near or below the HDL band may be removed with the gel rimming tool After correcting any gel tube artifacts rescan the gel template 011402 copy S amp S 3 Print Zoom in Zoom Out 3 6 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 www 4gc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS IMAGE ANALYSIS Introduction Lipoware is a Quantimetrix configured software program for the analysis of lipoprotein fract
36. e Lipoprint LDL System Table 1 2 19 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT EXPECTED VALUES Subfraction Cholesterol Cont Table 1 Normal population Midband Cholesterol LDL Subfraction Cholesterol VLDL Mid C Mid B Mid A LDL 1 LDL 2 LDL 3 Total LDL HDL TC mg dL mg dL mg dL mg dL mg dL mg dL mg dL mg dL mg dL mg dL range 6 26 9 24 5 17 6 26 24 59 4 32 0 4 59 128 40 103 123 199 mean 12 9 16 5 10 1 16 6 41 1 14 3 1 9 95 7 56 8 168 1 SD 4 12 2 82 2 40 4 26 7 85 6 82 0 81 16 56 11 29 18 45 pale 4 7 22 1 10 9 22 1 5 3 14 9 8 1 25 1 25 4 56 8 0 7 286 0 3 6 62 5 128 8 40 0 79 4 131 2 200 0 N 114 114 114 114 114 114 44 114 114 114 Number of samples showing the respective fractions It is recommended that each laboratory establish its own normal range which may be unique to the population it serves depending on geographical patient or environmental factors Data for the population outside the NCEP group N 141 aged 18 to 77 years 46 male and 54 female are shown in Table 2 Table 2 Population outside NCEP guidelines mg dL Midband Cholesterol LDL Subfraction Cholesterol VLDL Mid C Mid B Mid A LDL 1 LDL 2 LDL 3 LDL 4 LDL 5 Total LDL H
37. eaks Incomplete peak refer to page 3 19 Perform a manual analysis Bands migrated too far refer to page 3 19 Perform a manual analysis Misidentified HDL peak refer to page 3 19 Perform a manual analysis Noisy graphs dirty scans Dirty tubes Clean tubes rescan Too many trapped air bubbles Remove bubbles if possible and rescan or repeat test Shifted graph or unexpected peaks Chylomicron noisy peaks refer to pages 3 18 Perform a manual analysis Missing peaks refer to pages 3 17 Perform a manual analysis No color in peaks Misidentified peaks 4 5 Perform a manual analysis Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM TROUBLESHOOTING PRINTING AND TRASH PROBLEM POSSIBLE CAUSE SOLUTION No print out Printer connection error Check USB connection re start iMac and initiate printing No printer online Go to Apple Menu System Preferences Select printer and fax icon and select printer No ink Replace cartridge ON OFF Turn ON No paper Load paper tray Poor quality print Nozzle head dirty or out of alignment Refer to printer manual for cleaning instructions Low on ink Replace cartridge Trash will not empty Preview software is open Quit Preview and then go to Finder and Empty Tra
38. ect OK on the dialog box and try again clicking only once de improper band selection please try again Be 11 Improper band selection dialog box will appear by double clicking above VLDL for a second time The analysis will be terminated Select OK on the dialog box and close all windows Go back to page 3 21 to repeat the manual analysis IM Ab improper band selection analysis was BS terminated E The reanalyzed tube s are saved in the images folder on the desktop Follow detailed instructions on page 3 13 to view reanalyzed files Repeat manual reanalysis procedure until satisfactory reports are generated Re analysis of any samples using the same name will overwrite any previous report 3 24 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS IMAGE ANALYSIS Spread Sheet Report A tab delimited txt file with all numerical data will be created for every automated or manual analysis All txt files can be found in the images folder located on the desktop Each automated or manual analysis will create a new txt file To open the txt file double click on it To exit program select AppleWorks from the upper menu bar and choose Quit AppleWorks 3 25 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX
39. es 1 _ Append file extension tif V Send scanned images to Add ie New Folder Cancel Save A 3 4 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS SCANNING Operation Cont 4 A prompt will indicate scanning in progress ScanWizard Pro 4800 AF Scanner Context 4800 AF Scanner Job Positive Status Reading and processing image data _ _ Stop 5 After scanning is completed the program will return to the ScanWizard Pro start screen pg 3 2 Quit the ScanWizard Pro program by selecting ScanWizard Pro from the upper menu then choose Quit ScanWizard Pro QA About ScanWizard Pro 7 bo j ScanWizard Pro 4800 AF Scanner Overview Image le Scanner View Preferences Correction Help Preferences R T ome EA p steed Conon C meran C san gt amp ite 2 Hide ScanWizard Pro7 3H wea KEH Hide Others MESLE wi Output 8 000 x 10 000 Scaling 100 0 1 8 M8 i Transom Fo BP M Sampling None Category None i D Auto Corrections 5 D Range Automatic E WAB Points No Correction E Tone Cure no Comecon E BEC No Correct rection E Color No Correction Filters None E Aromatic Color Restoration f i i la 3 5 Quantimetrix 2005 Manhattan Beach Blvd Re
40. for HDL C LDL C sum of Midbands C B A and LDL subfractions and VLDL C are shown in Table 3 Table 3 Intra assay precision data for HDL LDL and VLDL sample N Mean mg l SD ee Meat nora SD J CN Menn nga 9D al 1 l 55 147 268 86 090 1 05 17 099 5 86 2 2 42 0 78 1 87 120 143 1 20 15 099 643 3 2 31 0 90 2 87 133 202 152 35 197 558 4 2 8 137 284 180 202 112 23 166 728 Precision Data For Midbands C B A and LDL subfractions 1 7 are seen in Tables 4 and 5 Table 4 Intra assay precision data for Midband subfractions Sample N Mean ngl SD OV o Mean mg d SD CV Mean g dl SD CV 1 12 16 076 4 75 9 0 60 6 67 14 156 11 14 2 12 17 144 847 13 051 3 92 14 072 514 3 lel 22 121 5 50 16 0 47 2 94 13 050 3 85 4 2 30 144 4 80 15 136 907 10 o77 7 70 2 21 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT SPECIFIC PERFORMANCE CHARACTERISTICS Table 5 Intra assay precision data for LDL subfractions LDL 1 SD CV LDL 2 SD CV LDL 3 SD CV LDL 4 SD CV LDL 5 SD CV LDL 6 SD CV LDL 7 SD CV Sample N Mean Mean Mean Mean Mean Mean Mean mg
41. ht 120V 220V Electrophoresis Chamber Power Supply 120V 220V Rimming Tool USB Printer Cable Tube Adapter User s Manual Part Catalog Number 48 7002 48 9530 48 9519 Part Catalog Number C0950 C0944 C0945 43 5301 43 5311 5312 43 6001 43 3903 3904 43 5451 C0952 43 5581 MO920 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM ONE YEAR WARRANTY Coverage This warranty covers any defective materials or workmanship with exceptions stated below with regard to the Lipoprint system product Duration The warranty remains in effect one year from the date of original purchase Not Covered This warranty does not cover damage caused by misuse or use other than intended and described in the product instruction manual Use of equipment other than that supplied by Quantimetrix voids this warranty The purchaser is responsible for costs of shipping customs duty and delivery regarding this warranty Quantimetrix will repair any defects in materials or workmanship not caused by purchaser or exchange the defective unit with a reconditioned or new unit Purchaser is responsible for set up of replacement instrumentation Extended Warranty The purchaser may extend all warranty terms an additional year by purchasing an extended warranty policy for 10 of the purchase price N B Purchaser means either distributor or e
42. ions cont b With the scanner turned OFF turn the scanner on its side to see the locking screw at the bottom Using a small screwdriver turn the locking screw counterclockwise to unlock it When successfully unlocked the screw will push out a little nearly even with the bottom of the scanner Note Always re lock scanner before moving or transporting 1 8 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM ABOUT THIS SYSTEM ASSEMBLY Lipoprint System Assembly Instructions cont c Use the USB cable to connect scanner to the iMac ch Warning Do not plug the scanner into the power source until the USB cable is securely attached The USB connection could be damaged if connected with the power ON AEA NS d Plug in the power cord to the back of the scanner and the other end into the surge protected power strip Power strip not included with international systems 4 Color printer a Unpack the printer and retain all packaging materials b Place the printer in a convenient location near the iMac computer 1 9 Quantimettrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 www 4gc com MO92002A 6 10 LIPOPRINT SYSTEM ABOUT THIS SYSTEM ASSEMBLY Lipoprint System Assembly Instructions cont c Attach the rounded end of the USB cable to the printer
43. ions and subfractions using the Lipoprint system Lipoware drops vertical lines at predefined Rf values separating the various lipoprotein bands It determines the area under the curve in area and calculates the cholesterol concentration in mg dL for each band The analysis program requires the inputs of a Total Cholesterol TC value for each sample to be analyzed The TC values must be obtained externally e g clinical analyzer Operation 1 Initiate analysis by clicking on the Lipoware Clinical icon in the dock 3 7 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS IMAGE ANALYSIS Operation Cont 2 Select Macros from the upper menu bar then drag down to Lipoprint LDL and click on it ee ete a Lee ee 3 A Open Image Files window will appear Select the template image to be analyzed located under Desktop Lipoprint Data Double click on the selected template image a gt ss jm L Lipoprint Data Zz Q search DEVICES images EE Test Template T a Administra K T Macintosh HD PLACES Applications lipousert gt MEDIA w Enable All Images C Open _ 3 8 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALY
44. ishandling of tube containers Discard gel tube s No band migration No power Check power supply connection No buffer in upper chamber Add buffer Air bubbles Dislodge bubbles No sample Repeat test Unusual migration rate Wrong current Check power setting Wrong buffer dilution Repeat buffer dilution Overused buffer Make new buffer Cold buffer Bring buffer to room temperature Poor band resolution Heparinized samples Do not use Deteriorated samples Refer to Specimen Collection in product insert Not enough sample or loading gel Check pipette calibration Cold buffer Bring buffer to room temperature Band above stacking gel Chylomicron presents in samples Use only fasting samples Aggregation by improperly frozen samples Refer to Specimen Collection in product insert 4 2 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM TROUBLESHOOTING SCANNING PROBLEM POSSIBLE CAUSE SOLUTION Scanner not responsive No connection to computer Check connections Scanner OFF Turn ON scanner Scanner cannot complete scan Template holder not pushed in completely Push in re scan Black template image Scanner set at reflective mode Rescan in positive transparency mode Negative template image Scanner set at negative mode Rescan in positive transparency mode Error Code 147 or 122 Template holder not pushed in comple
45. l scanner for gel tube image acquisition e Color printer for report generation 1 Lipoprint electrophoretic equipment and reagents are described in detail in Chapter 2 2 iMac computer a Unpack the iMac from its box Save all packing materials in the event that the system needs to be returned for repair b The computer has been preconfigured with all the software necessary to run the Lipoprint System c Attach the power cord into the back of the iMac screen Plug the iMac into the surge protected power strip provided Power strip not included with international systems Warning Keep the power strip in OFF position until all the components are connected d Place the keyboard in front of the iMac plug the USB cable from the keyboard into the USB port Ch in the rear of the iMac screen 1 6 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM ABOUT THIS SYSTEM ASSEMBLY Lipoprint System Assembly Instructions cont e Plug the USB cable from the mouse into any of the two USB ports on the keyboard 3 Digital scanner a Unpack the scanner and retain all packaging materials 1 7 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM ABOUT THIS SYSTEM ASSEMBLY Lipoprint System Assembly Instruct
46. lectrophoresis the lipoprotein particles are concentrated by the loading and stacking gels into a sharp narrow band As the lipoprotein particles migrate through the separating gel matrix they are resolved into lipoprotein bands according to their particle sizes from largest to smallest due to the sieving action of the gel HDL migrates the farthest followed by small dense LDL larger buoyant LDL Midbands comprising primarily IDL and VLDL Chylomicrons if present will appear above the stacking gel or remain in the loading gel A typical Lipoprint profile consists of 1 VLDL band 3 Midbands up to 7 LDL bands and 1 HDL band After the electrophoresis is completed the various stained lipoprotein fractions bands present in the sample are identified by their mobility Rf using VLDL as the starting reference point VLDL 0 and HDL as the leading point HDL 1 Fig 3 cathode Rf 0 00 VLDL 0 09 c 017 B Midband 0 27 A 0 32 1 Large LDL 0 38 0 45 3 oe 4 small LDL 0 56 5 0 60 6 0 64 7 Rf gt 1 00 HDL End of Gel anode amp Figure 3 Mobilities of the Lipoprotein Bands The relative area for each lipoprotein band is determined and multiplied by the total cholesterol concentration of the sample to yield the amount of cholesterol for each band in mg dL The total cholesterol concentration of the sample needs to be measured independently e g a clinical analyzer or a point of care instrument 2 5
47. lines for desirable lipid status LDL C is comprised of the sum of cholesterol in Mid bands C through A as well as all the subfractions Lipoprint Profile with unrecognized HDL peak HDL missing because of improper band recognition Solution Perform manual analysis as indicated in pages 3 21 to 3 24 Be sure to click slightly below HDL 3 18 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS REPORT GENERATION Flawed Report 5 SAMPLE tube 1 Quantimetrix LIPOPRINT SYSTEM SMEL o Omet LPO PRINT SYSTEM Sub fraction cholesterol 11 2 18 2 4 ee mgidl HI H fil Range Range LDL Profile not indicative of Type A presence of small dense LDL Total Chol mgfdl 100 2200 LDL Chol mgfdl 52 lt 130 Reference ranges derived from 125 serum samples that met the NCEP ATPIII puidelines for desirable lipid status LDL C is comprised of the sum of cholesterol in Mid bands C through A as well as all the subfractions Lipoprint Profile with distorted distribution Distorted distribution due to improper band selection Solution Perform manual analysis 3 19 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS REPORT GENERATION gr 1212006 SAMPLE
48. n classes are heterogeneous consisting of multiple subfractions that vary with respect to particle size density and chemical composition Lipoprotein heterogeneity has been demonstrated by various analytical methods such as density gradient ultracentrifugation 3 nuclear magnetic resonance NMR 4 non denaturing gradient gel electrophoresis GGE 5 and the Lipoprint System a linear polyacrylamide gel electrophoresis system 6 LDL can be resolved into a maximum of seven LDL subfractions with the Lipoprint System The LDL subfractions have been named LDL 1 consisting of the largest particles through LDL 7 consisting of the smallest particles Genetic as well as environmental factors are responsible for the differences in the degree of LDL heterogeneity among subjects Age gender and lipid status are known to affect the LDL subfraction profile 7 Individuals exhibiting lipoprotein profiles consisting primarily of the larger buoyant LDL 1 and LDL 2 subfractions have been designated as Pattern A while profiles with predominantly smaller and denser subfractions LDL 3 through LDL 7 have been designated as Pattern B 5 Fig 1 Normal VLDL IDL LDL 1 2 HDL Abnormal i VLDL IDL LDL 1 7 HDL Figure 1 Normal Pattern A and abnormal Pattern B lipoprotein profiles 2 2 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO9200
49. nd Type B abnormal based on the average particle size of the LDL particles 5 A normal Lipoprint profile with predominately large LDL LDL 1 and LDL 2 is consistent with Type A Fig 11 Chol mg dL 12 16 15 17 47 17 73 ange gs fora s2 13 5 80 eee Total LDL C mg dL 112 lt 130 LDL Profile Indicative of TYPE A mainly large LDL Total Chol mg dl 198 lt 200 Figure 11 Typical Normal Lipoprint Profile 2 16 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT QUANTITATIVE RESULTS An abnormal Lipoprint profile with predominantly small LDL LDL 3 to LDL 7 is consistent with Type B as described in the literature Fig 12 11 2 2001 SAMPLE 20 Q27a Quantimetrix Lipoprinit System 9 17 Area l fe 6 0 121133 9 0 81 4 9 1 1 15 3 Chol mg dL 31 26 16 31 34 23 21 13 3 maid HI HI HI HI HI HI HI HI Reference 22 13 15 25 57 30 Reference Range Range Total LDL C mg dL 189 HI 130 LDL Profile Not Indicative of TYPE A presence of small dense LDL Total Chol mg dL 260 HI 200 Figure 12 Typical abnormal Lipoprint profile 2 17 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT LIMITATIONS e The Lipoprint LDL test
50. nd user If a purchaser makes a purchase directly from Quantimetrix then Quantimetrix is responsible for the warranty If the purchaser makes a purchase from a distributor then the distributor must honor all warranty issues Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 e www 4gc com MO92002A 6 10
51. ng a wide range of cholesterol concentrations and containing 7 12 fractions and subfractions were kept in the refrigerator for 7 days The Lipoprint profiles for all samples were generated on day 3 and day 7 and the resulting cholesterol values of all lipoprotein fractions and subfractions N 206 were compared Cholesterol day 3 0 996 Cholesterol day o 0 075 r 0 976 Cholesterol day 7 0 964 Cholesterol day 0 0 758 r 0 961 It was concluded that serum and EDTA plasma are equally suitable as specimens and that the samples may be stored refrigerated 2 8 C for up to 7 days Accuracy by Correlation The Lipoprint Test System was compared to a direct HDL method EQUAL Diagnostics HDL Direct Liquid Select and a direct LDL method EQUAL Diagnostics LDL Direct Liquid Select A population of 268 serum samples with LDL C ranging from 55 218 mg dL LipoPrint and 54 215 mg dL direct LDL as well as HDL C ranging from 24 129 mg dL LipoPrint and 26 137 mg dL direct HDL was evaluated Table 15 and 16 Table 15 Lipoprint HDL Direct HDL N 268 268 mean mg dL 53 2 54 8 SD mg dL 15 13 15 43 regression Lipoprint HDL 0 9361 HDL giract 1 8607 2 0 912 Table 16 Lipoprint LDL Direct LDL N 268 268 mean mg dL 121 9 116 3 SD mg dL 30 73 29 58 regression Lipoprint LDL 0 998 LDL girect 5 7995 0 923 2 27 Quantimetrix 2005 Manhattan Beach Blvd
52. poprint System LDL Subfractions Template 1 Mark the center of each lipoprotein band present with a marking pen on the glass tube 2 Align the VLDL fraction at the top of the separating gel with the line on the template marked VLDL 3 Slide the gel along the template until the HDL band is superimposed over the line marked HDL 4 Determine the lipoprotein subfractions present in the sample by matching the bands on the gel to the corresponding lines on the template 2 15 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT QUANTITATIVE RESULTS Note A total cholesterol value for each sample to be analyzed must be obtained using a clinically approved cholesterol method prior to generating quantitative results The total cholesterol concentration of the sample must be gt 100mg dL The Gel Tubes are scanned and the relative area of each lipoprotein subfraction is established by dropping vertical lines at predetermined cut off ranges for each band The amount of cholesterol in each lipoprotein band is calculated by multiplying the relative area of each band by the total cholesterol of the sample LDL cholesterol is calculated as the sum of the cholesterol concentrations of all the LDL subfractions plus the Midbands A B and C Traditionally lipoprotein subfraction profiles have been classified into Type A normal a
53. port on the rear panel of the printer and the USB connector to the open port on the keyboard or on the back of the iMac d Plug the power cord into the surge protected power strip Power strip not included with international systems Now that the system assembly is completed turn ON the system in the following sequence 1 Power strip 7 2 iMac On button is located on the back of the screen J 3 Scanner wait for the scanner to complete it s initiation cycle 4 Printer It is recommended to turn OFF the Lipoprint System when not in use 1 10 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM ABOUT THIS SYSTEM LOGIN Upon turning on the iMac a Mac OSX window will appear with three different options e Administrator e Lipouser1 e Lipouser2 1 11 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM ABOUT THIS SYSTEM LOGIN Administrator Login Administrator has privileges to change settings on System Preferences Lipousers do not have such privileges 1 Double click on Administrator A new window will appear 2 Enter the password Password This password may be changed by the administrator 3 Amenu bar will appear at the top of the screen Clicking on the apple icon and selecting Sy
54. rix Corporation 2 13 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT QUALITATIVE RESULTS Note The various bands of a Lipoprint profile may be identified using the Lipoprint Template for a qualitative assessment homogeneous LDL distribution vs heterogeneous distribution The lipoprotein profile reflects the lipid status of the patient sample Fig 9 A normal lipoprotein profile Pattern A will typically exhibit bands for VLDL Midbands C B and A these comprise IDL LDL 1 and 2 and HDL The presence of additional LDL subfractions LDL 3 through 7 is indicative of heterogeneous LDL i e Pattern B Figure 9 Lipoprotein subfraction distributions for five individuals from a homogeneous LDL pattern on the left to a progressively more heterogeneous pattern on the right 2 14 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT QUALITATIVE RESULTS Using the Lipoprint Template The Lipoprint LDL System Template is used to identify the lipoprotein subfractions present on the gel Fig 10 Rf 0 00 me 0 09 0 17 027 0 38 c A 0 1 051 2 i 3 4 5 6 7 Rf gt 1 00 HDL lt End of Ge Figure 10 Li
55. sh 4 6 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM FOR RESEARCH USE ONLY The Lipoprint LDL System with the Lipoware Clinical analysis software has been cleared by the FDA for the analysis of lipoprotein subfractions in human serum or plasma Upon customer request for supporting ongoing research selected units of the Lipoprint LDL System have been modified with additional software for Research Use Only RUO The RUO software is not intended for diagnostic purposes as it has not been reviewed by the FDA The Lipoware LDL RUO and Lipoware HDL RUO icons located on the dock of the Lipoprint LDL System computer screen identify the Research Use Only analysis software In contrast the Lipoware Clinical icon identifies the FDA cleared analysis software IL RUO Lipoware Clinical Icon Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 www 4gc com MO92002A 6 10 LIPOPRINT SYSTEM PARTS AND ACCESSORIES PART NUMBER UPDATES Part numbers are subject to change Consult the Quantimetrix Web Site for updated information REAGENTS Description Lipoprint LDL Kit 100 Tests Liposure Lipoprotein Control Lipoprint System Buffer HARDWARE Description Digital Scanner iMac Computer Color Printer Preparation Rack Preparation Lig
56. should be used in conjunction with other data e g additional clinical testing physician observations family history etc e Heparin interferes with the separation of LDL subfractions e Total cholesterol in the sample must be above 100mg dL to avoid overestimation of VLDL cholesterol e Chylomicrons in sample will invalidate the subfraction cholesterol measurement Appearance of the specimen i e cloudiness turbidity or a creamy layer on top of the sample after overnight refrigeration should be used as the confirming test for presence of chylomicrons e Lipoprint results have not been evaluated for testing lipoproteins during pregnancy e NCEP has not published guidelines for optimal desirable values for LDL subfractions 2 18 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT EXPECTED VALUES HDL C and LDL C The National Cholesterol Education Program NCEP had established risk related cut off points for LDL cholesterol and HDL cholesterol These were recently reaffirmed in May 2001 by the third report on the NCEP Adult treatment Panel ATP III 10 Fraction Range Status HDL C 40 mg dL Decreased Risk at Higher Levels lt 40 mg dL Increased Risk LDL C lt 100 mg dL Optimal for Patients with Coronary Heart Disease CHD lt 130 mg dL Desirable 130 159 mg dL Borderline High Risk 160
57. solution in the lower chamber and 200mL in the upper chamber The lower buffer may be reused up to five times Use only fresh buffer in the upper chamber The buffer must be at room temperature RT 18 27 C 10 After both chambers are assembled and filled with buffer thoroughly examine each tube for air bubbles Dislodge any bubbles with a pipettor tip Bubbles could obstruct the passage of electrical current 2 11 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM LIPOPRINT LDL KIT ASSAY PROCEDURE 11 Put the electrophoresis chamber lid in place and connect it to the power supply Fig 8 Adjust the power supply to deliver the current of 3mA per Gel Tube e g 36mA for 12 tubes 18mA for 6 tubes etc The voltage should be set at a maximum delivery of 500V Figure 8 Assembled chamber 12 Electrophoresis time is approximately 60 minutes Stop the electrophoresis when the HDL fraction is about lcm from the bottom of the fastest migrating Gel Tube 13 When the electrophoresis is complete turn the power OFF remove the chamber lid and discard the electrolyte buffer in the upper chamber The lower buffer may be retained and reused up to 5 times Discard after 7 days 14 Before removing the Gel Tubes from the electrophoresis chamber wipe off excess buffer then place in the Preparation Rack for transport to the sc
58. stem Preferences will give access to all computer systems OOO System Preferences Lale Showa Personal lo is lO eo iu F E D lie lo ia m fe fe do fie Je R e 2 Startup Disk Time Machine 1 12 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM ABOUT THIS SYSTEM LOGIN Lipouser Login 1 Double click on Lipouser or Lipouser2 A new window will appear 2 Enter the password user Only the administrator has privileges to change the password 3 The Lipouser desktop includes the following a Desktop icons e Macintosh HD e Images folder e Lipoprint Data folder b Dock icons e Finder e Dashboard e Safari e Appleworks e ScanWizard Pro e Lipoware Clinical e Printer e Preview e Trash 1 13 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM C h a pte r 2 LIPOPRINT LDL KIT TOPICS SEE PAGE Intended Use and Indications For Use 2 1 Summary and Explanation of the Test 2 2 Test Principle 2 4 Product Description 2 6 Warnings and Precautions 2 8 Specimens and Specimen Collection 2 8 Assay Procedure 2 9 Quality Control 2 13 Qualitative Results 2 14 Quantitative Results 2 16 Limitations 2 18 Expected Values 2 19 Specific Performance Characteristics 2
59. ted quit Lipoware by selecting Lipoware Clinical on upper menu bar A window asking Cancel or Continue will appear Ignore this warning all information has already been saved Select Continue to proceed toon aa Ue TEN 3 12 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS REPORT GENERATION 1 To view the report of previously analyzed gel tubes double click the images folder on the desktop 2 To select one report at a time double click on the individual PICT icon To select all the reports at once marquis select by clicking on the outer left side of the icons Click hold and drag the marquis box to highlight all reports Double click on any highlighted icon to open reports To select reports a la carte hold down the Apple Command key while highlighting the specific file icon Double click on any highlighted icon to open selected reports Note The Lipoprint report window may be enlarged by clicking on the Zoom In icon located above the image profile Click once on the green button in the upper left hand side of the profile window to enlarge the window To close the opened image profile click once on the red x button located in the upper left hand side of the window Clicking on the yellow button in the upper left hand side of the window will position the file at the dock Cli
60. tely Push in re scan 4 3 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM TROUBLESHOOTING ANALYSIS PROBLEM Cut off template image POSSIBLE CAUSE Pulled tray out before scan is completed SOLUTION Rescan scanning completed when Start Screen reappears Macro error copy failed error message Tube entry exceeded maximum Relaunch enter correct of tubes Array out of index error message HDL bands have migrated too far Perform a manual analysis 1 0 Error code 120 Software corruption Missing Images folder on desktop Call Technical Support Error code 48 Duplicate file name Close program relaunch Lipoware Error code 107 Memory overload too many applications opened on desktop Close all applications relaunch Lipoware Macro error window not found in line 1114 1139 Operator clicked with the mouse on the desktop while software is generating profiles Close program re launch Lipoware Error code 5000 Image folder is locked Unlock image folder 4 4 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM TROUBLESHOOTING REPORT GENERATION PROBLEM POSSIBLE CAUSE SOLUTION No color in HDL p
61. x 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM ABOUT THIS SYSTEM PRINCIPLES OF OPERATION Lipoprint LDL is an electrophoresis system capable of separating lipoprotein fractions and subfractions The resolved lipoprotein bands are scanned analyzed and a report is generated Electrophoresis Lipoproteins are stained with a lipophilic dye and separated using a high resolution linear polyacrylamide gel Scan The gel tubes are placed into the digital scanner to generate an image of the resolved lipoprotein bands Analysis The digital image is analyzed using Lipoware software which calculates the cholesterol level for each of the lipoprotein fractions and subfractions Report An easy to interpret color graph of the patient s lipoprotein profile is generated and printed 1 5 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM ABOUT THIS SYSTEM ASSEMBLY Lipoprint System Assembly Instructions Clear a convenient space for setting up the computer scanner and printer The space should be about 3 X 6 with adequate ventilation and a nearby power outlet The Lipoprint system is comprised of 4 functional components e Lipoprint electrophoretic equipment and reagents e iMac computer for data acquisition and analysis e Digita
62. ylomincrons bubbles or artifacts appear in VLDL area shifting VLDL to Mid C area Solution Perform manual analysis as indicated in pages 3 21 to 3 24 Be sure to click center of tube slightly above VLDL excluding chylomicrons bubbles or any other artifact 3 15 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS REPORT GENERATION Flawed Report 2 SAMPLE tube 1 Quantimetrix LIPOPRINT SYSTEM i HDL Sub fraction cholesterol 15 5 10 10 10 0 8 8 mgfdl HI HI HI Rane p2 a noas a eoo o parang LDL Profile not indicative of Type A presence of small dense LDL Total Chol gdi 100 2200 LDL Chol mg dl 66 lt 130 Reference ranges derived from 125 serum samples that met the NCEP ATEI pens for desirable lipid status LDL C is comprised of the sum of cholesterol in Mid bands C through A as well as all the subfractions Lipoprint Profile shifted to the left VLDL peak missing while Mid band C appear in VLDL area Solution Perform manual analysis as indicated in pages 3 21 to 3 24 Be sure to click center of tube slightly above VLDL 3 16 Quantimetrix 2005 Manhattan Beach Blvd Redondo Beach CA 90278 1205 USA e 1 310 536 0006 FAX 1 310 536 9977 e www 4qc com MO92002A 6 10 LIPOPRINT SYSTEM SAMPLE ANALYSIS REPORT GENERATION Flawed Report 3 at 1212006 SAMPL

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lipoprint&reg; system user's manual

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lipoprint® system user's manual