HUMAN IgG SUBCLASS LIQUID REAGENT KITS (for use on the
Contents
1. Insert Code SIN083 Version 12 August 2010 Page 7 of 10 10 12 ans IgG1 46 6421 4230 10600 lgG2 46 1980 760 3550 lgG3 46 620 173 1730 lgG4 46 323 16 1150 Total IgG 46 10200 5680 14900 12 14 ans IgG1 54 6581 3420 11500 lgG2 54 2486 1000 4550 lgG3 54 583 283 1250 lgG4 54 409 37 1360 Total IgG 54 10500 6640 14900 14 18 ans IgG1 48 6057 3150 8550 IgG2 48 2614 640 4950 IgG3 48 708 230 1960 IgG4 48 521 110 1570 Total IgG 48 10100 5500 14400 12 Les performances indiqu es ci dessous sont des donn es g n r es sur l automate BN II de Siemens qui fonctionne de la m me fa on que le BN ProSpec 12 1 Pr cision intra essai Trois s rums contenant des taux diff rents de sous classes d IgG ont t test s 10 fois Toutes les concentrations sont exprim es en mg L IgG1 IgG2 IgG3 IgG4 Moy CV Moy CV Moy CV Moy CV S rum 1 40529 1 56 8256 2 59 668 6 2 74 534 1 3 05 Serum 2 12630 1 67 1736 2 16 517 8 2 76 396 1 2 95 Serum 3 3350 6 44 758 5 8 19 353 1 3 49 324 8 5 08 12 2 Precision inter essai Trois s rums contenant des taux diff rents de sous classes d IgG ont t test s au cours de trois s ries diff rentes Toutes les concentrations sont exprim es en mg L N nombre d essais IgG1 IgG2 IgG3 IgG4 Moy CV Moy CV Moy CV Moy CV S rum 1 40765 had 7811 s 6972. 4 1 Antis rums lgG1 lgG2 lls sont monosp cifiques pour la sous classe humaine appropri e lls sont fournis sous forme liquide et contiennent 0 099 d azide de sodium 0 1 d acide E amino n caproique EACA 0 5 BSA et 0 01 benzamidine comme conservateurs 4 2 R actif I9G3 IgG4 Ils sont monosp cifiques pour la sous classe appropri e et sont fournis sous forme iquide Ils contiennent 0 033 d azide de sodium 0 05 ProClin 0 1 EACA et 0 01 benzamidine comme conservateurs 4 3 Calibrateurs et controles Ils sont pr par s partir de pools de s rum humains et sont fournis sous forme liquide et stable lls contiennent 0 099 d azide de sodium 0 1 d EACA et 0 01 de benzamidine comme conservateurs La concentration en sous classes d IgG indiqu e sur les tiquettes des flacons ont t obtenues par comparaison avec la r f rence internationale CRM 470 r f 5 44 R actif suppl mentaire Il contient 0 099 d azide de sodium comme conservateur ProClin est une marque depos e par Rohm and Haas Corp Philadelphia PA 5 PRECAUTIONS Tous les s rums humains fournis dans ce coffret ont t test s et trouv s n gatifs pour l antigene de surface de l h patite B Ag HBs pour le virus de l h patite C et pour les anticorps anti virus de Pimmunod ficience humaine HIV1 and HIV2 Les tests utilis s ont soit t approuv s par la FDA USA ou accept pour un usage en diagnostic in vitro par l union europ enne Di
3. Drei Serum Pr parationen mit unterschiedlichen IgG Subklassen Konzentrationen wurden 10 mal gemessen Alle Konzentrationsangaben sind in mg L IgG1 IgG2 IgG3 IgG4 Mittelwert VK Mittelwert VK Mittelwert VK Mittelwert VK Serum 1 40529 1 56 8256 2 59 668 6 2 74 534 1 3 05 Serum 2 12630 1 67 1736 2 16 517 8 2 76 396 1 2 95 Mixing normal Start wash 06 goto waste End wash 06 park needle Incubation time 1 10sec 0 Incubation delta 1 10sec O Medium 1 Type SystemLiquid Medium ID Diluens Syringe profile 7 Volumel ul 160 Medium 2 Type SystemLiquid Medium ID Air Bubble Syringe profile 2 Volumelul 2 Medium 3 Type Reagent Medium ID 1304 G4L No Calib Syringe profile 3 Volume ul 50 Medium 4 Type Destination Medium ID Syringe profile 6 Volumelul 112 8 5 Messbereich und Sensitivit t Ungef hrer Messbereich Ungef hre Sensitivit t Spezifit t Probenver Probenver mg l d nnung mg l d nnung IgG1 2625 84000 1 100 131 1 5 IgG2 613 19600 1 20 153 1 5 IgG3 55 875 1 100 2 7 1 5 IgG4 38 613 1 100 1 9 1 5 Serum 3 3350 6 44 758 5 8 19 353 1 3 49 324 8 5 08 9 INTERPRETATION DER ERGEBNISSE Qualit tskontrolle Die IgG Subklassen Konzentrationen der Kontrollen sind auf dem jeweiligen Flaschenetikett angegeben Der von dem BN ProSpec gemessene Wert sollte maximal 15 vom angegebenen Wert abwe
4. Medium ID Air Bubble Volume pl 2 Medium ID 1303 G3L Volume ul 50 Medium ID Volume pl 112 Name G4L Assayclass Neph Sample dilution 1 100 Overflow 0 End wash 00 normal F No Calib End wash 06 park needle No Calib long name TBS LigG4 Assaygroup Serum Min dilution 1 5 Measurement transfer 2 Meas window 1 10 sec from 75 until 3600 Calibrator 1306 IgG Std Start dilution 1 80 Supporting points 5 Dilution Deviation 1 100 15 1 100 15 Insert Code SIN083 Version 12 August 2010 Page 9 of 10 Level Medium Low Transfers 1 Mixing No mixing Start wash 06 goto waste End wash 00 normal Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Medium ID Diluens Syringe profile 7 Volumel ul 160 Medium 2 Type Reagent Medium ID 1305 SRL Syringe profile 2 Volume ul 10 Medium 3 Type Sample Medium ID No Calib Syringe profile 3 Volume pl Medium 4 Type Destination Medium ID ringe profile 3 Transfers 2 Mixing normal Start wash 06 goto waste End wash 06 park needle Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Medium ID Diluens Syringe profile 7 Volumel ul 160 Medium 2 Type SystemLiquid Medium ID Air Bubble Syringe profile 2 Volume pl 2 Medium 3 Type Reagent Medium ID 1304 G4L No Calib Syringe prof
5. HUMAN IgG SUBCLASS LIQUID REAGENT KITS for use on the Siemens BN ProSpec Analyser For in vitro diagnostic use Product code LK001 P NK006 P NK007 P LK008 P LK009 P Product manufactured by The Binding Site Group Ltd PO Box 11712 Birmingham B14 4ZB U K www bindingsite co uk Telephone 44 0 121 436 1000 Fax 44 0 121 430 7061 e mail info bindingsite co uk BN ProSpec is a registered trademark of Siemens Healthcare Diagnostics Inc 1 INTENDED USE This kit is intended for quantifying human IgG subclasses 1 2 3 and 4 immunoglobulins in serum on the BN ProSpec Measurement of these immunoglobulins aids in the diagnosis of abnormal protein metabolism and the body s lack of ability to resist infectious agents 2 SUMMARY AND EXPLANATION In normal adults IgG constitutes approximately 75 of the total serum immunoglobulin Within the IgG class the usual order of concentration of the 4 subclasses is IgG 1 gt lgG2 gt IgG3 gt IgG4 but the actual concentration of each may vary markedly between individuals The four IgG subclasses show considerable differences in their properties including ability to fix complement to bind to macrophages and to pass through the placenta Abnormal levels of one or more subclass may be associated with certain conditions including anaphylaxis autoimmune and gut diseases as well as hypo and hyper gammaglobulinaemia ref 1 In particular reduced production of IgG2 in chi
6. goto waste Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type SystemLiquid Syringe profile 2 Medium 3 Type Sample Syringe profile 3 Medium 4 Type Destination Syringe profile 3 End wash 00 normal Medium ID Buffer Volumel ul 170 Medium ID Air Bubble Volume pl 2 Medium ID Volume pl No Calib Medium ID Volume pl Transfers 2 Mixing normal Start wash 06 goto waste Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type SystemLiquid Syringe profile 2 Medium 3 Type Reagent Syringe profile 3 Medium 4 Type Destination Syringe profile 6 End wash 06 park needle Medium ID Buffer Volumel ul 170 Medium ID Air Bubble Volume pl 2 Medium ID 1301 G1 Volume ul 40 No Calib Medium ID Volumel ul 112 Version 100 Result unit mg L Evaluation Measurement duration 1 10 sec 3600 Evaluation method 8 Two point BNII Blockmode 0 Calibration Calibration method LogitLog Deviation 5 Cal valid until 100 Special parameters Parameter 1 2 Controls Control ID Name 0776 TBS IgG Sc 1 0975 TBS IgG Sc 2 Transfers 1 Mixing No mixing Incubation time 1 10sec O Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type SystemLiquid Syringe profile 2 Medium 3 Type Sample Syr
7. tre conserv s entre 2 et 8 C et peuvent tre utilis s jusqu la date de p remption indiqu e sur l tiquette de la bo te Les r actifs non ouverts calibrateurs et contr les doivent tre gard s 2 8 C Apr s ouverture ils sont stables quatre semaines 2 8 C s ils ne sont pas contamin s ou vapor s Il est recommand de ne pas laisser les flacons d antis rum ouverts sur les n ph l m tres temp rature ambiante en dehors de leur utilisation Au cours du temps les flacons d antis rums non ouverts peuvent d velopper un pr cipit ou un trouble qui n est d une contamination bacterienne et qui n affecte pas l activit de l antiserum Dans de telles situations l antis rum doit tre clarifi par filtration travers un filtre ayant des pores de 0 2 um ou par centrifugation approximativement 10 minutes 15000g 7 PRELEVEMENT ET PREPARATION DE L ECHANTILLON Utiliser des s rums fraichement pr lev s ou congel s Les pr l vements sanguins doivent tre collect s par ponction veineuse le caillot doit se former naturellement et le s rum doit tre r cup r le plus rapidement possible pour viter l h molyse Le s rum peut tre stock 8 jours 2 8 C avant le test ou 20 C ou une temp rature inf rieure pour un stockage prolong du s rum non dilu Les cong lations et d cong lations successives peuvent abimer le s rum Les s rums lipidiques h molys s comtamin s par des micr
8. 0975 TBS IgG Sc 2 1 20 Low long name TBS IgG2 Assaygroup Serum Min dilution 1 5 Calibrator 1306 IgG Std Start dilution 1 5 Supporting points 6 Insert Code SIN083 Version 12 August 2010 Page 1 of 10 Transfers 1 Mixing No mixing Incubation time 1 10sec 0 Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type SystemLiquid Syringe profile 2 Medium 3 Type Sample Syringe profile 3 Medium 4 Type Destination Syringe profile 3 Transfers 2 Mixing normal Incubation time 1 10sec 0 Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type SystemLiquid Syringe profile 2 Medium 3 Type Reagent Syringe profile 3 Medium 4 Type Destination Syringe profile 6 IgG3 Assay Details Assay ID 203 Version 100 Result unit mg L Evaluation Measurement duration 1 10 sec 3600 Evaluation method 8 Two point BNI Blockmode 0 Calibration Calibration method LogitLog Deviation 5 Cal valid until 100 Special parameters Parameter 1 2 Controls Control ID Name 0776 TBS IgG Sc 1 0975 TBS IgG Sc 2 Transfers 1 Mixing No mixing Incubation time 1 10sec O Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type Reagent Syringe profile 2 Medium 3 Type Sample Syringe profile 3 Medium 4 Type Destination Syringe profile 3 Transfers 2 Mixing normal Incubation time 1 10sec O Medium 1 Type SystemLiquid Sy
9. 1 Mixing No mixing Start wash 06 goto waste End wash 00 normal Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Medium ID Buffer Syringe profile 7 Volume ul 170 Medium 2 Type SystemLiquid Medium ID Air Bubble Syringe profile 2 Volume ul 2 Medium 3 Type Sample Medium ID No Calib Syringe profile 3 Volumef ul Medium 4 Type Destination Medium ID Syringe profile 3 Volumef ul Transfers 2 Mixing normal Start wash 06 goto waste End wash 06 park needle Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Medium ID Buffer Syringe profile 7 Volume ul 170 Medium 2 Type SystemLiquid Medium ID Air Bubble Syringe profile 2 Volume pl 2 Medium 3 Type Reagent Medium ID 1301 G1 F No Calib Syringe profile 3 Volume ul 40 Medium 4 Type Destination Medium ID Syringe profile 6 Volume pl 112 1962 Assay Details Assay ID 202 Name G2 Version 100 Assayclass Neph Result unit mg L Sample dilution 1 20 Evaluation Measurement duration 1 10 sec 3600 Measurement transfer 2 Evaluation method 8 Two point BNII Meas window 1 10 sec from 75 until 3600 Blockmode 0 Overflow 0 Calibration Calibration method LogitLog Deviation 5 Cal valid until 100 Special parameters Parameter 1 2 Controls Control ID Name Dilution Deviation Level 0776 TBS IgG Sc 1 1 20 Medium
10. 160 Medium 2 Type SystemLiquid Medium ID Air Bubble Syringe profile 2 Volume pl 2 Medium 3 Type Reagent Medium ID 1304 G4L No Calib Syringe profile 3 Volume ul 50 Medium 4 Type Destination Medium ID Syringe profile 6 Volume pl 112 8 5 Gamme de mesure et sensibilite Gamme de mesure Sensibilit Specificite Dilution Dilution ais chantillon ent echantillon lgG1 2625 84000 1 100 131 1 5 IgG2 613 19600 1 20 153 1 5 IgG3 55 875 1 100 2 7 1 5 lgG4 38 613 1 100 1 9 1 5 9 INTERPRETATION DES RESULTATS Contr le de qualit Les concentrations des contr les sont indiqu es sur l tiquette du flacon Les r sultats seront corrects si la valeur obtenue diff re au maximum de 15 par rapport la valeur cible 10 LIMITES 10 1 Il existe une bonne corr lation entre les dosages n ph l m triques et les dosages en immunodiffusion radiale et ceci avec un grand nombre d chantillons 10 2 Cependant les performances de ce coffret ne peuvent tre garanties pour des chantillons contenant des concentrations lev es en paraprot ines ou complexes Version 100 Result unit mg L Assayclass Neph Sample dilution 1 100 Assaygroup Serum Min dilution 1 5 immuns circulants facteur rhumato de ou pour des s rums h molys s ou lipidiques Ces s rums ne peuvent tre dos s par cette technique cause de la d viation de Evaluation Measurement duration 1 10 sec 360
11. 06 goto waste Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type SystemLiquid Syringe profile 2 Medium 3 Type Reagent Syringe profile 3 Medium 4 Type Destination Syringe profile 6 End wash 06 park needle Medium ID Buffer Volumelul 170 Medium ID Air Bubble Volume pl 2 Medium ID 1301 G1 Volumelul 40 No Calib Medium ID Volume pl 112 Name G2 Assayclass Sample diluti Version 100 Result unit mg L Evaluation Measurement duration 1 10 sec 3600 Evaluation method 8 Two point BNII Blockmode 0 Calibration Calibration method LogitLog Deviation 5 Cal valid until 100 Special parameters Parameter 1 2 Controls Control ID Name Dilution 0776 TBS IgG Sc 1 1 20 0975 TBS IgG Sc 2 1 20 Transfers 1 Mixing No mixing long name TBS IgG2 Neph Assaygroup Serum on 1 20 Min dilution 1 5 Measurement transfer 2 Meas window 1 10 sec from 75 until 3600 Overflow 0 Calibrator 1306 IgG Std Start dilution 1 5 Supporting points 6 Deviation 15 15 Start wash 06 goto waste End wash 00 normal Incubation time 1 10sec 0 Incubation delta 1 10sec O Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type SystemLiquid Syringe profile 2 Medium 3 Type Sample Syringe profile 3 Medium 4 Type Destination Syringe profile 3 Medium ID Vo
12. 5 K 532 6 O S rum 2 12402 A 1621 er 525 9 4 387 7 Rod S rum 3 3043 Hi 780 1 353 1 as 319 6 vs BN est une marque d pos e de Siemens Healthcare Diagnostics Inc 13 BIBLIOGRAPHIE 1 Shakib F 8 Stanworth D R 1980 Human IgG subclasses in health and disease Part 11 La Ricerca Clin Lab 10 561 580 2 Shackleford P G Polmar S H et al 1986 Spectrum of IgG2 subclass deficiency in children with recurrent infections Prospective study J Pediatr 108 647 653 3 Hamilton R H 1987 Human IgG subclass measurements in the clinical laboratory Clin Chem 33 1707 1725 4 Schur PH 1987 IgG subclasses a review Annals of Allergy 58 89 99 5 Carr Smith H D Overton J Bradwell A R 1997 IgG subclass value assignment to the protein reference preparation CRM470 Clin Chem Vol 43 No 56 PS238 KITS REACTIVOS L QUIDOS SUBCLASES IgG HUMANA para uso en el analizador Siemens BN ProSpec Para uso diagn stico in vitro nicamente Codigo LK001 P NK006 P NK007 P LK008 P LK009 P Producto fabricado por The Binding Site Group Ltd PO Box 11712 Birmingham B14 4ZB U K www bindingsite co uk The Binding Site Spain S L U C Balmes 243 4 32 08006 Barcelona Tel fono 902027750 Fax 902027752 e mail info bindingsite es web www bindingsite es BN ProSpec e una marca registrada de Siemens Healthcare Diagnostics Inc 11 UTILIZACI N E
13. Control Human IgG Subclass High Control 4 x 1 6mL antiserum or 4 x 2 4mL Latex Reagent to the appropriate subclass 2 x 3 0mL BS Supplementary Reagent L in LK008 P and LK009 P kits only a go m co 0 MO No 1 p amp w D Ed D Materials required but not provided Equipment for collection and preparation of test samples A fully operational and equipped Analyser mm 99 mo o GD Bo amp Hib ND O1 BR D 8 4 Test procedure 8 4 1 The user should be familiar with the operation of the BN ProSpec analyser before attempting to carry out the test procedures 8 4 2 Prepare the analyser as per the user s manual 8 4 3 The IgG subclass assay protocols are defined in the analyser 8 4 4 A Siemens representative is required to create the new assay protocols Assay Details Assay ID 201 Name G1 Version 100 Assayclass Neph Result unit mg L Sample dilution 1 100 Evaluation Measurement duration 1 10 sec 3600 Measurement transfer 2 Evaluation method 8 Two point BNII Meas window 1 10 sec from 75 until 3600 Blockmode 0 Overflow 0 Calibration Calibration method LogitLog Deviation 5 Cal valid until 100 Special parameters Parameter 1 2 Controls Control ID Name Dilution Deviation 0776 TBS IgG Sc 1 1 100 15 0975 TBS IgG Sc 2 1 100 15 long name TBS IgG1 Assaygroup Serum Min dilution 1 5 Calibrator 1306 IgG Std Start dilution 1 10 Supporting points 6 Transfers
14. Erkrankungen in Ver bindung gebracht werden z B Autoimmunerkrankungen Anaphylaxie Darmerkrankungen sowie Hypo und Hypergammaglobulin mien Ref 1 Speziell bei Kindern kann sich eine verminderte IgG2 Produktion durch wiederkehrende Infektionen bemerkbar machen Ref 2 Reviews Ref 3 4 Das BN ProSpec erlaubt eine vollautomatische schnelle und quantitative Bestimmung der Pr zipitationsreaktion zur Proteinbestimmung 3 TESTPRINZIP Die Nephelometrie beruht auf der Messung von Streulicht das durch l sliche Partikel in einer L sung erzeugt wird Dieses Prinzip kann f r die Messung von l slichen Antigenen angewendet werden da sie mit dem entsprechenden spezifischen Antik rper reagieren und einen unl slichen Antigen Antik rper Komplex Immunkomplex bilden Diese Immunkomplexe streuen in die L sung eingestrahltes Licht und wenn der Antik rper im berschuss vorliegt ist die Lichtstreuung direkt proportional zur Antigenkonzentration Durch Messung eines verschieden verd nnten Standards mit bekannter Antigenkonzentration wird eine Kalibrationskurve erstellt Die Antigenkonzentration von unbekannten Proben wird nach Messung direkt an Hand der Kalibrationskurve ermittelt Latex verst rkte Antik rper Bei einigen Antigen Antik rper Reaktionen werden nicht gen gend gro e Immunkomplexe gebildet und diese k nnen daher nicht nephelometrisch bestimmt werden Ist der Antik rper an ein Latexpartikel mit geeigneter Gr e gekoppelt
15. Seren oder h molytische oder lip mische Seren verwenden 8 TESTDURCHF HRUNG 8 1 Gelieferte Materialien f r Kombi Kit Bestell Nr LK001 P 8 1 1 2x 1mL BS IgG Sub Std Human IgG Subklassen Kalibrator 8 1 2 2 x 1mL BS IgG Sub Low Control Human IgG Subklassen Kontrolle Low Level 8 1 3 2 x 1mL BS IgG Sub Control Human IgG Subklassen Kontrolle High Level 8 1 4 1x 1 6mL Human IgG1 BS Antiserum IgG1 Subklassen Antiserum 8 1 5 1 x 1 6mL Human IgG2 BS Antiserum IgG2 Subklassen Antiserum 8 1 6 1 x2 4mL 9G3 BS Latex IgG3 Latex Reagenz 8 1 7 1 x2 4mL gG4 BS Latex IgG4 Latex Reagenz 8 1 8 1 x 3mL BS Supplementary Reagent L BS Supplementary Reagent L f r die IgG3 und IgG4 Tests 8 2 Bestell Nr NK006 P NK007 P LK008 P LK009 P 8 2 1 2x 1mL BS IgG Sub Std Human IgG Subklassen Kalibrator 8 2 2 2 x 1mL BS IgG Sub Low Control Human IgG Subklassen Kontrolle Low Level 8 2 3 2 x 1mL BS IgG Sub Control Human IgG Subklassen Kontrolle High Level 8 2 4 4 x 1 6mL Antiserum oder 4 x 2 4mL Latexreagenz der jeweiligen Subklasse 8 2 5 2 x 3mL BS Supplementary Reagent L nur f r LK008 P und LK009 P 8 3 Ben tigte nicht im Kit enthaltene Materialien 8 3 1 Laborausstattung zum Sammeln und Vorbereiten der Proben Probenr hrchen Zentrifugen etc 8 3 2 Einen voll ausgestatteten und funktionsf higen BN ProSpec 8 4 Testdurchf hrung 8 4 1 Der Anwender sollte mit dem BN ProSpec vertraut sein bevor der Test dur
16. Si quedan fuera de los l mites aceptables en un 15 frente a la nueva curva de calibraci n se ha de comprobar el instrumento antes de repetir el ensayo Si los problemas contin an p ngase en contacto con su proveedor 10 4 Los resultados de la medida de subclases IgG no deben utilizarse para evaluar atopia en pacientes al rgicos 10 5 No debe realizarse el diagn stico ni iniciarse un tratamiento bas ndose nicamente en la medida de las subclases IgG deben tenerse en cuenta tambi n la historia cl nica y resultados de otras pruebas de laboratorio 11 VALORES ESPERADOS Los valores indicados a continuaci n se obtuvieron a partir de un n mero limitado de muestras inglesas y se ofrecen nicamente como una gu a Es muy recomendable que cada laboratorio establezca sus propios rangos de normalidad 11 1 Rangos normales adultos Estos rangos se obtuvieron midiendo las subclases en sueros proporcionados por el Birmingham Blood Transfusion Service de donantes adultos sanos Subclase N mero Media Mediana Rango 95 IgG n mg L mg L Percentil mg L IgG1 30 6330 6085 3824 9286 IgG2 30 4528 4541 2418 7003 IgG3 30 790 7 706 4 218 2 1760 6 19G4 30 280 0 215 3 39 2 864 0 11 2 Rangos pedi tricos Estos rangos se obtuvieron midiendo la IgG total y las subclases en muestras de suero pedi tricas del Birmingham Hospital utilizando productos de inmunodifusi n radial de Bi
17. and ProClin 300 and must be handled with caution suitable gloves and other protective clothing should be worn at all times when handling this product Do not ingest or allow contact with the skin particularly broken skin or open wounds or mucous membranes If contact does occur wash with a large volume of water and seek urgent medical advice Explosive metal azides may be formed on prolonged contact of sodium azide with lead and copper plumbing on disposal of reagent flush with a large volume of water to prevent azide build up This product should only be used by suitably trained personnel for the purposes stated in the Intended Use Strict adherence to these instructions is essential at all times Results are likely to be invalid if parameters other than those stated in these instructions are used Reagents from different batch numbers of kits are NOT interchangeable If large numbers of tests are performed care should be taken to ensure that all the reagents are from the same batch 6 STORAGE AND STABILITY The unopened kits should be stored at 2 8 C and can be used until the expiry date given on the kit box label Unopened reagent calibrators and controls should be stored at 2 8 C Once opened they are stable for four weeks at 2 8 C providing precautions to prevent evaporation and contamination are taken Open vials should not be left on the analyser when they are not in use Over time unopened vials of antisera can develop a preci
18. from 75 until 3600 Overflow 0 Calibrator 1306 IgG Std Start dilution 1 80 Supporting points 5 Deviation Level 15 Medium 15 Low Transfers 1 Mixing No mixing Start wash 06 goto waste End wash 00 normal Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Medium ID Diluens Syringe profile 7 Volumel ul 160 Medium 2 Type Reagent Medium ID 1305 SRL Syringe profile 2 Volume ul 10 Medium 3 Type Sample Medium ID No Calib Syringe profile 3 Volumef ul Medium 4 Type Destination Medium ID Syringe profile 3 Volume pl Transfers 2 8 10 Jahre gG1 32 6345 4320 10200 gG2 32 2126 720 4300 gG3 32 521 127 853 gG4 32 349 19 932 Total IgG 32 9700 5680 13600 10 12 Jahre gG1 46 6421 4230 10600 gG2 46 1980 760 3550 IgG3 46 620 173 1730 IgG4 46 323 16 1150 Total IgG 46 10200 5680 14900 12 14 Jahre gG1 54 6581 3420 11500 gG2 54 2486 1000 4550 gG3 54 583 283 1250 gG4 54 409 37 1360 Total IgG 54 10500 6640 14900 14 18 Jahre gG1 48 6057 3150 8550 gG2 48 2614 640 4950 gG3 48 708 230 1960 gG4 48 521 110 1570 Total IgG 48 10100 5500 14400 12 LEISTUNGSDATEN Die unten aufgef hrten Leistungsdaten wurden auf einem Siemens BN II ermittelt der nach dem gleichen Prinzip wie der BN ProSpec arbeitet ermittelt 12 1 Intra Assay Variation
19. 0 8420 Medium 2 IgG2 39 838 225 3000 Type SystemLiquid Medium ID Air Bubble IgGs 39 445 186 853 Syringe profile 2 Volume pl 2 IgG4 39 129 5 784 Medium 3 Total IgG 39 6640 3270 12700 Type Reagent Medium ID 1303 G3L No Calib 2 4 ans Syringe profile 3 Volumef ul lgG1 36 5495 3150 9450 Medium 4 IgG2 36 1146 360 2250 Type Destination Medium ID IgG3 36 396 173 676 Syringe profile 6 Volumef ul 19G4 36 171 10 537 Total IgG 36 7770 4680 12500 IgG4 4 6 ans Assay Details IgG1 49 6149 3060 9450 Assay ID 204 Name G4L long name TBS LigG4 IgG2 49 1496 605 3450 Version 100 Assayclass Neph Assaygroup Serum IgG3 49 531 99 1221 Result unit mg L Sample dilution 1 100 Min dilution 1 5 IgG4 49 329 18 1125 Evaluation Total IgG 49 9280 5320 13400 je uroment en pro EN een os i 5 eh 6 8 ans valuation method 8 Two poin leas window sec from until Blockmode 0 P Overflow 0 IgG 43 5758 2880 9180 Calibration IgG2 43 1754 440 3750 Calibration method LogitLog Calibrator 1306 IgG Std IgG3 43 437 155 853 Deviation 5 Start dilution 1 80 IgG4 43 331 4 992 Cal valid until 100 Supporting points 5 Total IgG 43 8860 4540 13600 Special parameters 8 10 ans at 1 2 IgG1 32 6345 4320 10200 Control ID Name Dilution Deviation Level es En ee 0776 TBS IgG Sc 1 1 100 15 Medium g 0975 TBS IgG Sc 2 1 100 15 Low IgG4 32 349 19 932 Total IgG 32 9700 5680 13600
20. 0 Evaluation method 8 Two point BNI Blockmode 0 Calibration Calibration method LogitLog Deviation 5 Cal valid until 100 Special parameters Parameter 1 2 10 4 Controls Control ID Name Dilution lumi re non sp cifique intrins que ce type d chantillons Des r sultats surprenants Measurement transfer 2 doivent tre confirm s par une autre m thode Meas window 1 10 sec from 75 until 3600 Overflow 0 10 3 Il est fortement recommand aux utilisateurs de passer les deux contr les lors de chaque serie Si une valeur obtenue partir d une courbe de calibration enregistr e est en dehors des valeurs attendues la calibration doit tre refaite Si les valeurs des controles pr sentent une variation supp rieure 15 par rapport aux valeurs attendues v rifier l instrument avant de r p ter l essai Si le probl me persiste contacter le fournisseur Calibrator 1306 IgG Std Start dilution 1 80 Supporting points 5 Les r sultats du dosage des sous classes ne doivent pas tre utilis s chez les sujets uk allergiques lors de maladies atopiques Deviation Level 0776 TBS IgG Sc 1 1 100 15 Medium 10 5 Le diagnostic et le traitement ne peuvent pas tre uniquement bas s sur le dosage des 0975 TBS IgG Sc 2 1 100 15 Low sous classes d IgG La clinique et les autres r sultats du laboratoire doivent tre pris en compte Transfers 1 11 VALEURS ATTENDUES Mixing No mixing Start wash 06
21. 0sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Medium ID Buffer Syringe profile 7 Volume ul 170 Medium 2 Type SystemLiquid Medium ID Air Bubble Syringe profile 2 Volume ul 2 Medium 3 Type Reagent Medium ID 1301 G1 No Calib Syringe profile 3 Volume ul 40 Medium 4 Type Destination Medium ID Syringe profile 6 Volume pl 112 Insert Code SIN083 Version 12 August 2010 Page 6 of 10 Assay ID 202 Version 100 Result unit mg L Evaluation Measurement duration 1 10 sec 3600 Evaluation method 8 Two point BNI Blockmode 0 Calibration Calibration method LogitLog Deviation 5 Cal valid until 100 Special parameters Parameter 1 2 Controls Control ID Name Dilution 0776 TBS IgG Sc 1 1 20 0975 TBS IgG Sc 2 1 20 Name G2 Transfers 1 Mixing No mixing Assayclass Neph Sample dilution 1 20 long name TBS IgG2 Assaygroup Serum Min dilution 1 5 Measurement transfer 2 Meas window 1 10 sec from 75 until 3600 Overflow 0 Calibrator 1306 IgG Std Start dilution 1 5 Supporting points 6 Deviation 15 15 Start wash 06 goto waste End wash 00 normal Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type SystemLiquid Syringe profile 2 Medium 3 Type Sample Syringe profile 3 Medium 4 Type Destination Syringe profile 3 Medium ID
22. 1 10sec O Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type Reagent Syringe profile 2 Medium 3 Type Sample Syringe profile 3 Medium 4 Type Destination Syringe profile 3 Medium ID Volumef ul Medium ID Volumef ul Medium ID Volumef ul Medium ID Volumeful Transfers 2 Mixing normal Diluens 160 1305 SRL 10 F No Calib Start wash 06 goto waste End wash 06 park needle Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type SystemLiquid Syringe profile 2 Medium 3 Type Reagent Syringe profile 3 Medium 4 Type Destination Syringe profile 6 Medium ID Volumef ul Volumef ul Medium ID Volumef ul Medium ID Volumef ul Name G4L Assayclass Sample diluti Version 100 Result unit mg L Evaluation Measurement duration 1 10 sec 3600 Evaluation method 8 Two point BNII Blockmode 0 Calibration Calibration method LogitLog Deviation 5 Cal valid until 100 Special parameters Parameter 1 2 Controls Control ID Name Dilution 0776 TBS IgG Sc 1 1 100 0975 TBS IgG Sc 2 1 100 Insert Code SIN083 Version 12 August 2010 Page 4 of 10 Diluens 160 Medium ID Air Bubble 2 1303 G3L No Calib 50 112 long name TBS LlgG4 Neph Assaygroup Serum ion 1 100 Min dilution 1 5 Measurement transfer 2 Meas window 1 10 sec
23. 3 IgG4 reagent These are monospecific for the relevant subclass and are supplied in stabilised liquid form They contain 0 033 sodium azide 0 05 ProClin 0 1 EACA and 0 01 benzamidine as preservatives 4 3 Calibrators and controls These consist of pooled human serum and are supplied in stabilised liquid form They contain 0 099 sodium azide 0 1 EACA and 0 01 benzamidine as preservatives The concentration of IgG subclasses given on the vial label has been obtained by comparison with the CRM470 international reference material ref 5 4 4 Supplementary reagent Contains 0 099 sodium azide as a preservative ProClin is a trademark of Rohm and Haas Corp Philadelphia PA 5 CAUTION All donors of human serum supplied in this kit have been serum tested and found negative for hepatitis B surface antigen HBsAg and antibodies to human immunodeficiency virus HIV1 and HIV2 and hepatitis C virus The assays used were either approved by the FDA USA or cleared for in vitro diagnostic use in the EU Directive 98 79 EC Annex ll however these tests cannot guarantee the absence of infective agents Proper handling and disposal methods should be established as for all potentially infective material including but not limited to users wearing suitable protective equipment and clothing at all times Only personnel fully trained in such methods should be permitted to perform these procedures WARNING This product contains sodium azide
24. 40 8420 gG2 39 838 225 3000 gG3 39 445 186 853 gG4 39 129 5 784 Total IgG 39 6640 3270 12700 2 4 years gG1 36 5495 3150 9450 gG2 36 1146 360 2250 gG3 36 396 173 676 gG4 36 171 10 537 Total IgG 36 7770 4680 12500 4 6 years gG1 49 6149 3060 9450 gG2 49 1496 605 3450 gG3 49 531 99 1221 gG4 49 329 18 1125 Total IgG 49 9280 5320 13400 6 8 years gG1 43 5758 2880 9180 IgG2 43 1754 440 3750 gG3 43 437 155 853 gG4 43 331 4 992 Total IgG 43 8860 4540 13600 8 10 years gG1 32 6345 4320 10200 gG2 32 2126 720 4300 gG3 32 521 127 853 gG4 32 349 19 932 Total IgG 32 9700 5680 13600 10 12 years gG1 46 6421 4230 10600 gG2 46 1980 760 3550 gG3 46 620 173 1730 IgG4 46 323 16 1150 Total IgG 46 10200 5680 14900 12 14 years gG1 54 6581 3420 11500 gG2 54 2486 1000 4550 gG3 54 583 283 1250 gG4 54 409 37 1360 Total IgG 54 10500 6640 14900 Insert Code SIN083 Version 12 August 2010 Page 2 of 10 14 18 years IgG1 48 6057 3150 8550 IgG2 48 2614 640 4950 IgG3 48 708 230 1960 IgG4 48 521 110 1570 Total IgG 48 10100 5500 14400 12 PERFORMANCE CHARACTERISTICS The performance characteristics shown here is data generated on a Siemens BN II which is an analyser that performs in the same way as the BN ProSpec 12 1 Within run precision Three serum preparations containing different
25. Volume pl Medium ID Volume pl Medium ID Volumef ul Medium ID Volume ul Transfers 2 Mixing normal Buffer 170 Air Bubble 2 No Calib Start wash 06 goto waste End wash 06 park needle Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type SystemLiquid Syringe profile 2 Medium 3 Type Reagent Syringe profile 3 Medium 4 Type Destination Syringe profile 6 Medium ID Volumef ul Medium ID Volumef ul Medium ID Volume ul Medium ID Volumef ul 1963 Assay Details Assay ID 203 Name G3L Buffer 170 Air Bubble 2 1302 G2 No Calib 40 112 long name TBS LlgG3 Transfers 1 Mixing No mixing Start wash 06 goto waste End wash 00 normal Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type Reagent Syringe profile 2 Medium 3 Type Sample Syringe profile 3 Medium 4 Type Destination Syringe profile 3 Medium ID Diluens Volume ul 160 Medium ID 1305 SRL Volume ul 10 No Calib Medium ID Volume pl Medium ID Volumef ul Transfers 2 Mixing normal Start wash 06 goto waste End wash 06 park needle Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Medium ID Diluens Syringe profile 7 Volume ul
26. associ s certaines pathologies comme l anaphylaxie les maladies autoimmunes digestives les hypo et hypergammaglobulinemies r f 1 En particulier une diminution de synth se de I lgG2 chez l enfant peut tre associ e des infections r currentes r f 2 Cet aspect a t bien tudi ref 3 et 4 Le n ph l m tre BN ProSpec permet le dosage rapide des prot ines sp cifiques par n ph l m trie 3 PRINCIPE La n ph l m trie permet la mesure de la lumi re diffus e par des particules en solution Ceci peut tre appliqu la mesure d un antigene soluble si en presence d un anticorps appropri des immun complexes insolubles sont form s et dispersent la lumi re Lorsque l anticorps est en exc s a lumi re diffus e est directement proportionnelle la concentration en antigene Une courbe de calibration est tablie en utilisant une s rie de calibrateurs dilu s dont la concentration en antig ne est connue La concentration des s rums inconnus est alors calcul e partir de la courbe de calibration R actif latex Au cours de certaines r actions antig nes anticorps les complexes form s ne sont pas suffisamment gros pour tre d tect s en n ph l m trie Si l anticorps est coat sur des particules de latex de taille suffisante la capacit de diffraction des complexes immuns form s avec l antigene est alors suffisamment augment e pour permettre la d tection n ph l m trique 4 REACTIFS
27. ayaron 3 preparaciones de suero con niveles distintos de subclases IgG Cada una se ensay 10 veces Todas las concentraciones est n en mg L IgG1 IgG2 IgG3 IgG4 Media CV Media CV Media CV Media CV Suero 1 40529 1 56 8256 2 59 668 6 2 74 534 1 3 05 Suero 2 12630 1 67 1736 2 16 517 8 2 76 396 1 2 95 Suero 3 3350 6 44 758 5 8 19 353 1 3 49 324 8 5 08 12 2 Precisi n inter ensayo Se ensayaron 3 preparaciones de suero con niveles distintos de subclases IgG al menos en tres ensayos distintos Todas las concentraciones est n en mg L N n mero de ensayos IgG1 IgG2 IgG3 IgG4 E CV 3 CV A CV i CV Media Media Media Media N N N N 4 38 6 84 4 41 4 10 1 407 7 7811 N 697 5 1 532 6 hi Suero 0765 6 8 6 3 3 2 39 4 18 2 38 4 13 Suero 2 12402 6 1621 6 525 9 4 387 7 3 6 95 2 43 3 00 4 38 4 i 780 1 7 353 1 i 319 6 i Suero 3 3043 6 80 6 3 4 BN e una marca registrada de Siemens Healthcare Diagnostics Inc 13 BIBLIOGRAFIA 1 Shakib F amp Stanworth D R 1980 Human IgG subclasses in health and disease Part 11 La Ricerca Clin Lab 10 561 580 2 Shackleford P G Polmar S H et al 1986 Spectrum of IgG2 subclass deficiency in children with recurrent infections Prospective study J Pediatr 108 647 653 3 Hamilton R H 1987 Human IgG subclass measurements in the clinical laboratory Clin C
28. chgef hrt wird 8 4 2 Das Ger t wie im Handbuch des Herstellers beschrieben vorbereiten 8 4 3 Die IgG Subklassen Testparameter werden im Ger t angelegt 8 4 4 F r die Programmierung neuer Testparameter ist ein Siemens Mitarbeiter erforderlich IgG1 Assay Details Assay ID 201 Version 100 Result unit mg L Evaluation Measurement duration 1 10 sec 3600 Evaluation method 8 Two point BNI Blockmode 0 Calibration Calibration method LogitLog Deviation 5 Cal valid until 100 Special parameters Parameter 1 2 Controls Control ID Name Dilution Deviation Level 0776 TBS IgG Sc 1 1 100 15 Medium 0975 TBS IgG Sc 2 1 100 15 Low Name G1 Assayclass Neph Sample dilution 1 100 long name TBS IgG1 Assaygroup Serum Min dilution 1 5 Measurement transfer 2 Meas window 1 10 sec from 75 until 3600 Overflow 0 Calibrator 1306 IgG Std Start dilution 1 10 Supporting points 6 Transfers 1 Mixing No mixing Start wash 06 goto waste Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type SystemLiquid Syringe profile 2 Medium 3 Type Sample Syringe profile 3 Medium 4 Type Destination Syringe profile 3 End wash 00 normal Medium ID Buffer Volumel ul 170 Medium ID Air Bubble Volume pl 2 Medium ID Volume ul No Calib Medium ID Volumef ul Transfers 2 Mixing normal Start wash
29. cuerpo adecuado Se hace pasar un haz de luz a trav s del recipiente de reacci n donde a medida que tiene lugar la reacci n ant geno anticuerpo se forman complejos inmunes que dispersan la luz Cuando el anticuerpo est en exceso la luz dispersa es directamente proporcional a la concentraci n de ant geno Se prepara inicialmente una curva de calibraci n con una serie de calibradores diluidos de concentraci n de ant geno conocida De este modo pueden ensayarse muestras de concentraci n desconocida de ant geno y determinarse dicha concentraci n utilizando la curva de calibraci n Anticuerpos mejorados con l tex Algunas reacciones ant geno anticuerpo no forman complejos inmunes suficientemente grandes para ser detectados por nefelometr a Si el anticuerpo se une a part culas de l tex de un tama o adecuado la capacidad del complejo inmune formado para dispersar la luz queda incrementada de modo que permite su detecci n nefelom trica 4 REACTIVOS 4 1 Antisueros lgG1 lgG2 humana Son monoespecificos para la subclase correspondiente y se suministran en forma l quida estabilizada Contienen azida s dica 0 099 BSA 0 5 cido E amino N caproico EACA 0 1 y benzamidina 0 01 como conservantes 4 2 Reactivo lgG3 IgG4 humana Son monoespecificos para la subclase correspondiente y se suministran en forma l quida estabilizada Contienen 0 033 azida s dica 0 05 ProClin 0 01 EACA y 0 01 benzamidina como conser
30. entrifugaci n unos 10 minutos a 15 000 x g 7 OBTENCI N Y CONSERVACI N DE MUESTRAS Utilice muestras de suero frescas o ultra congeladas Las muestras de sangre se han de obtener por punci n en vena dejar que coagule de modo natural y separar el suero lo antes posible para prevenir la hem lisis El suero se conserva a 2 8 C hasta 8 d as o se congela sin diluir a 20 C para periodos m s largos Evitar la congelaci n repetida de las muestras No deben utilizarse muestras hemolizadas lip micas con contaminaci n microbiana o muestras que contengan part culas 8 METODOLOGIA 8 1 Material suministrado c digo LK001 P kit 8 1 1 2 x 1mL BS IgG Sub Std Calibrador IgG subclase humana 8 1 2 2 x 1mL BS IgG Sub Low Control Control bajo IgG subclase humana 8 1 3 2 x 1mL BS IgG Sub Control Control IgG subclase humana 8 1 4 1x 1 6mL Human IgG1 BS Antiserum Antisuero de lgG1 8 1 5 1x 1 6mL Human lgG2 BS Antiserum Antisuero de IgG2 8 1 6 1 x2 4mL gG3 BS Latex L tex de lgG3 8 1 7 1 x 2 4mL 9G4 BS Latex L tex de I9G4 8 1 8 1x 3mL BS Supplementary Reagent L BS Reactivo suplementario L para los ensayos IgG3 e IgG4 8 2 C digo NK006 P NK007 P LK008 P and LK009 P kits 8 2 1 2x 1mL BS IgG Sub Std Calibrador IgG subclase humana 8 2 2 2 x 1mL BS IgG Sub Low Control Control bajo IgG subclase humana 8 2 3 2 x 1mL BS IgG Sub Control Control elevado IgG subclase humana 8 2 4 4 x 1 6mL antisuero o 4 x 2 4mL reacti
31. goto waste End wash 00 normal Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Medium ID Diluens Syringe profile 7 Volumel ul 160 Medium 2 11 1 Adultes Type Reagent Medium ID 1305 SRL Les valeurs ci dessous ont t obtenues partir d une population limit e de sujets sains britanniques Elles sont donn es titre indicatif Il est fortement conseill chaque laboratoire d tablir ses propres normes f la Sous classes Nombre Moyenne Mediane Gamme 95 ae Deli 2 IgG n mg L mg L percentile mg L Type Sample Medium ID No Calib IgG1 30 6330 6085 3824 9286 Syringe profile 3 Volumeful lgG2 30 4528 4541 2418 7003 Medium 4 lgG3 30 790 7 706 4 218 2 1760 6 Type Destination Medium ID IgG4 30 280 0 215 3 39 2 864 0 Syringe profile 3 Volume pl 112 Enfants Transfers 2 Ces valeurs ont t obtenues en mesurant les IgG totales et les sous classes en utilisant des Mixing normal Start wash 06 goto waste End wash 06 park needle s rums p diatriques provenant de l h pital de Birmingham Les r sultats ci dessous ont t obtenus Incubation time 1 10sec 0 Incubation delta 1 10sec 0 en immunodiffusion radiale mg L Toutes les concentrations sont en mg L Medium 1 i Type SystemLiquid Medium ID Diluens 0 2ans n Moyenne Gamme 95 percentilo Syringe profile 7 Volumel ul 160 IgG 39 5248 194
32. hem 33 1707 1725 Schur P H 1987 IgG subclasses a review Annals of Allergy 58 89 99 Carr Smith H D Overton J Bradwell A R 1997 IgG subclass value assignment to the protein reference preparation CRM470 Clin Chem Vol 43 No 56 PS238 a Insert Code SIN083 Version 12 August 2010 Page 10 of 10
33. ichen 10 GRENZEN DES TESTS 10 1 Die nephelometrische Bestimmung der IgG Subklassen korreliert bei einer gro en Probenzahl sehr gut zur Binding Site IgG Subklassen Single Dilution RID Methode 10 2 Dieser Kit ist nicht f r die Bestimmung von lip mischen oder h molysierten Proben oder Proben die Rheumafaktoren Paraproteine oder andere zirkulierende Immunkomplexe enthalten geeignet da diese Proben einen nicht vorhersagbaren Anteil an unspezifischen Streulicht erzeugen k nnen Ungew hnliche Ergebnisse sollten mit einer alternativen Methode berpr ft werden 10 3 Es wird dringend empfohlen dass bei jedem Testansatz beide Kontrollen mitgef hrt werden Wird eine gespeicherte Kalibrationskurve verwendet und eine der Kontrollen liegt nicht im Vertrauensbereich sollte der Test neu kalibriert werden Weicht die Kontrolle weiterhin mehr als 15 vom Sollwert ab sollte vor einer erneuten Testdurchf hrung das Ger t berpr ft werden Kann das Problem so nicht gel st werden wenden Sie sich bitte an ihre Lieferfirma 10 4 Die IgG Subklassen Ergebnisse sollten nicht zur Diagnose einer Atopie bei Allergie Patienten verwendet werden 10 5 Die Diagnose und Einleitung einer Therapie d rfen nicht ausschlie lich auf der IgG Subklassen Bestimmung basieren Das klinische Bild und andere serologische Befunde m ssen ebenfalls ber cksichtigt werden 11 ERWARTETE WERTE 11 1 Erwachsene Die unten aufgef hrten Normalbereiche basieren auf de
34. ile 3 Volume pl Medium 4 Type Destination Medium ID Syringe profile 6 Volume pl 8 5 Rango de medida y sensibilidad Rango de medida aproximada Sensibilidad aproximada Especificidad Diluci n de Diluci n de mg L muestra mg L muestra IgG1 2625 84000 1 100 131 1 5 IgG2 613 19600 1 20 153 1 5 IgG3 55 875 1 100 2 7 1 5 IgG4 38 613 1 100 1 9 1 5 9 INTERPRETACION DE DATOS Control de calidad Las concentraciones de subclases IgG asignadas a los controles suministrados se indican en las etiquetas de los viales Los valores obtenidos para el control suministrado deben estar dentro de 15 del valor de concentraci n indicado en la etiqueta 10 LIMITACIONES 10 1 Se ha comprobado que los ensayos nefelom tricos de subclases IgG correlacionan bien con los ensayos RID de diluci n nica de Binding Site con un amplio n mero de muestras 10 2 Estos kits no son adecuados para la medida de muestras con niveles elevados de factores reumatoides paraproteinas complejos inmuno circulantes CICs o para muestras hemolizadas o lip micas debido al impredecible grado de dispersi n inespec fica que pueden generar estas muestras Resultados inesperados deben confirmarse mediante un m todo alternativo 10 3 Se recomienda a los clientes que ensayen ambos controles con cada lote de muestras ensayadas Si un valor de control queda fuera de rango cuando se ensaya frente a una curva almacenada el ensayo debe recalibrarse
35. inge profile 3 Medium 4 Type Destination Syringe profile 3 Transfers 2 Mixing normal Incubation time 1 10sec O Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type SystemLiquid Syringe profile 2 Medium 3 Type Reagent Syringe profile 3 Medium 4 Type Destination Syringe profile 6 Assay Details Assay 03 Version 100 Result unit mg L Evaluation Measurement duration 1 10 sec 3600 Evaluation method 8 Two point BNII Blockmode 0 Calibration Calibration method LogitLog Deviation 5 Cal valid until 100 Special parameters Parameter 1 2 Controls Control ID Name 0776 TBS IgG Sc 1 0975 TBS IgG Sc 2 Transfers 1 Mixing No mixing Incubation time 1 10sec O Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type Reagent Syringe profile 2 Medium 3 Type Sample Syringe profile 3 Medium 4 Type Destination Syringe profile 3 Transfers 2 Mixing normal Incubation time 1 10sec O Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type SystemLiquid Syringe profile 2 Medium 3 Type Reagent Syringe profile 3 Medium 4 Type Destination Syringe profile 6 Version 100 Result unit mg L Evaluation Measurement duration 1 10 sec 3600 Evaluation method 8 Two point BNII Blockmode 0 Calibration Calibration method LogitLog Deviation 5 Cal valid until 100 Special parameters Parameter 1 2 C
36. iqu en Angleterre par la soci t The Binding Site Group Ltd PO Box 11712 Birmingham B14 4ZB U K www bindingsite co uk Distribu s en France par la soci t The Binding Site France 14 rue des Glairaux BP226 38522 Saint Egreve Cedex T l phone 04 38 02 19 19 Fax 04 38 02 19 20 e mail infoObindingsite fr BN ProSpec est une marque d pos e de Siemens Healthcare Diagnostics Inc 1 Ce coffret est destin au dosage quantitatif des sous classes 1 2 3 et 4 d IgG dans le s rum humain par n ph l m trie en utilisant le n ph l metre BN ProSpec de la soci t Siemens La mesure de ces immunoglobulines apporte une aide dans le diagnostic du m tabolisme anormal des prot ines et des probl mes de l organisme r sister aux agents infectieux 2 PRESENT ATION GENERALE Chez les sujets adultes en bonne sant l IgG repr sente environ 75 de la concentration en Immunoglobulines s riques totales Dans la classe des IgG les concentrations de chaque sous classe sont g n ralement dans l ordre 1gG1 gt 1gG2 gt 1gG3 gt lgG4 bien que la concentration effective de chaque sous classe puisse varier consid rablement selon les individus Les quatre sous classes d IgG pr sentent des diff rences consid rables dans leurs propri t s incluant la capacit de fixation du compl ment de se lier aux macrophages et de passer la barri re placentaire Des taux inusuels d une ou plusieurs sous classes peuvent tre
37. ir Bubble Syringe profile 2 Volume pl 2 Medium 3 Type Reagent Medium ID 1304 G4L No Calib Syringe profile 3 Volume ul 50 Medium 4 Type Destination Medium ID Syringe profile 6 Volume ul 112 8 5 Measuring range and sensitivity PES Approx measuring range Approx sensitivity IE mg L Sample dilution mg L Sample dilution IgG1 2625 84000 1 100 131 1 5 IgG2 613 19600 1 20 153 1 5 IgG3 55 875 1 100 2 7 1 5 IgG4 38 613 1 100 1 9 1 5 9 INTERPRETATION OF DATA Quality Control The IgG subclass concentration assigned to the controls are given on the bottle labels Results obtained during the run should only be accepted if they are within 15 of the value s stated 10 LIMITATIONS 10 1 Nephelometric IgG Subclass assays have been shown to correlate well versus Binding Site Single Dilution RID assays with a wide range of samples 10 2 These kits are not suitable for the measurement of samples containing rheumatoid factor paraproteins other circulating immune complexes CIC s or for lipaemic or haemolysed samples due to the unpredictable degree of non specific scatter these sample types may generate Unexpected results should be confirmed using an alternative assay method 10 3 Customers are strongly recommended to run both controls with every batch of samples being assayed Should a control value be out of range against a stored curve it is recommended that the assay be recalibrated Where con
38. kann der so geformte Immunkomplex ausreichend Licht streuen Jetzt ist das Antigen der nephelometrischen Bestimmung zug nglich 4 REAGENZIEN 4 1 Human _IgG1 IgG2 Antiserum Diese Antiseren sind monospezifisch f r die jeweilige Subklasse und liegen als stabilisierte Fl ssigkeit vor Enthaltene Konservierungsmittel 0 099 Natriumazid 0 1 E aminocaprons ure EACA 0 5 BSA und 0 01 Benzamidin 4 2 Human lgG3 IgG4 Latex Reagenz Diese Latex Reagenzien sind monospezifisch f r die jeweilige Subklasse und liegen als stabilisierte Fl ssigkeit vor Enthaltene Konser vierungsmittel 0 033 Natriumazid 0 05 ProClinTM 0 1 EACA und 0 01 Benzamidin 4 3 Kalibratoren und Kontrollen Sie werden aus gepooltem Humanserum hergestellt und liegen als stabilisierte Fl ssigkeiten vor Enthaltene Konservierungsmittel 0 099 Natriumazid 0 1 EACA und 0 01 Benzamidin Der Wert von IgG Subklassen wurde durch Vergleich mit dem internationalen CRM470 Referenzmaterial ermittelt Ref 5 4 4 B S Zusatzreagenz Enthaltene Konservierungsmittel 0 099 Natriumazid ProClin ist ein Warenzeichen von Rohm and Haas Corp Philadelphia PA 5 WARNUNGEN UND VORSICHTSMABNAHMEN Jede Einzelspende von humanem Serum wurde untersucht und bez glich Antik rper gegen das Human Immunschw che Virus HIV 1 amp 2 das Hepatitis C Virus und gegen das Hepatitis B Ober fl chenantigen HBsAG als negativ befunden Die hierf r verwendeten Te
39. l 3600 Calibrator 1306 IgG Std Start dilution 1 80 Supporting points 5 Dilution Deviation 1 100 15 1 100 15 Start wash 06 goto waste Incubation delta 1 10sec 0 Medium ID Diluens Volumel ul 160 Medium ID 1305 SRL Volume ul 10 Medium ID Volumef ul Medium ID Volume ul Start wash 06 goto waste Incubation delta 1 10sec O Medium ID Diluens Volumel ul 160 Medium ID Air Bubble Volume ul 2 Medium ID 1303 G3L Volumelul 50 Medium ID Volume pl 112 Name G4L Assayclass Neph Sample dilution 1 100 Overflow 0 End wash 00 normal No Calib End wash 06 park needle No Calib long name TBS LlgG4 Assaygroup Serum Min dilution 1 5 Measurement transfer 2 Meas window 1 10 sec from 75 until 3600 Calibrator 1306 IgG Std Start dilution 1 80 Supporting points 5 Dilution Deviation 1 100 15 1 100 15 Start wash 06 goto waste Incubation delta 1 10sec 0 Medium ID Diluens Volumel ul 160 Medium ID 1305 SRL Volume ul 10 Medium ID Volumef ul Medium ID Volume pl End wash 00 normal No Calib Transfers 2 Mixing normal Start wash 06 goto waste End wash 06 park needle Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Medium ID Diluens Syringe profile 7 Volume pl 160 Medium 2 Type SystemLiquid Medium ID A
40. l o las mucosas especilamente si hay heridas En caso de contacto lave con abundante agua y consulte a un m dico Con el plomo y el cobre pueden formarse azidas met licas explosivas Cuando se elimine el reactivo lave con mucha agua los recipientes para evitar la acumulaci n de azida El presente producto debe ser utilizado por personal especializado Se recomienda observar estrictamente el procedimiento indicado No se garantizan resultados v lidos obtenidos utilizando par metros diferentes que los indicados Los reactivos de diferentes lotes NO son intercambiables En caso de realizar un n mero elevado de tests averigue que todos los reactivos sean del mismo lote Insert Code SIN083 Version 12 August 2010 Page 8 of 10 6 CONSERVACION Y ESTABILIDAD El kit sin abrir debe conservarse a 2 8 C y puede utilizarse hasta la fecha de caducidad indicada en la etiqueta El reactivo calibradores y controles deben conservarse a 2 8 C Una vez abiertos son estables durante 4 semanas a 2 8 C siempre que se evite la evaporaci n y contaminaci n Los viales abiertos no deben dejarse en el analizador cuando no se est n utilizando Los viales de antisuero no abierto pueden con el tiempo desarrollar un precipitado o turbidez que no est originada por contaminaci n microbiana y que no afecta a la actividad del antisuero En estos casos el antisuero ha de aclararse por filtraci n a trav s de un filtro desechable de 0 2um de poro o por c
41. ldren may be associated with recurrent infections ref 2 The subject has been reviewed refs 3 4 The BN ProSpec permits fully automatic rapid quantitative measurement of light scattering reactions for protein determination 3 PRINCIPLE OF THE ASSAY Nephelometry involves the measurement of light scattered by particles in solution This can be applied to the measurement of soluble antigen since in the presence of the appropriate antibody insoluble immune complexes are formed which are light scattering When the antibody is in excess the light scatter is directly proportional to the concentration of antigen A calibration curve is initially prepared using a series of diluted standards of known antigen concentration Samples of unknown antigen concentration can then be assayed and their concentrations determined using the calibration curve Latex enhanced Antibodies Some antigen antibody reactions do not form sufficiently large immune complexes to be detected nephelometrically If the antibody is coated onto latex particles of a suitable size the light scattering ability of the immune complexes formed with antigen is enhanced sufficiently to enable nephelometric detection 4 REAGENTS 4 1 Human IgG1 IgG2 antisera These are monospecific for the relevant subclass and are supplied in stabilised liquid form They contain 0 099 sodium azide 0 1 E amino n caproic acid EACA 0 5 BSA and 0 01 benzamidine as preservatives 4 2 Human IgG
42. levels of IgG subclasses were assayed 10 times All concentrations are in mg L IgG SUBKLASSEN HUMAN KIT zur Verwendung auf dem Siemens BN ProSpec IgG1 IgG2 IgG3 IgG4 Mean CV Mean CV Mean CV Mean CV Serum 1 40529 1 56 8256 2 59 668 6 2 74 534 1 3 05 Serum 2 12630 1 67 1736 2 16 517 8 2 76 396 1 2 95 Serum 3 3350 6 44 758 5 8 19 353 1 3 49 324 8 5 08 12 2 Between run precision Three serum preparations containing different levels of IgG subclasses were assayed on at least three separate occasions All concentrations are in mg L N number of runs IgG1 IgG2 IgG3 IgG4 Mean ns Mean ne Mean F e Mean ne Serum1 40765 Ni 7811 i 697 5 43 532 6 er Serum2 12402 4 1621 Fr 525 9 e 387 7 aL Serum3 3043 780 1 Fi 353 1 ci 319 6 BN is a trademark of Siemens Healthcare Diagnostics Inc 13 BIBLIOGRAPHY 1 Shakib F amp Stanworth D R 1980 Human IgG subclasses in health and disease Part Il La Ricerca Clin Lab 10 561 580 2 Shackleford P G Polmar S H et al 1986 Spectrum of IgG2 subclass deficiency in children with recurrent infections Prospective study J Pediatr 108 647 653 3 Hamilton R H 1987 Human IgG subclass measurements in the clinical laboratory Clin Chem 33 1707 1725 4 Schur PH 1987 IgG subclasses a review Annals of Allergy 58 89 99 5 Carr Smith H D Overton J Brad
43. lumef ul Volumef ul Medium ID Volumef ul Medium ID Volumef ul Transfers 2 Mixing normal Buffer 170 Medium ID Air Bubble 2 No Calib Start wash 06 goto waste End wash 06 park needle Incubation time 1 10sec 0 Incubation delta 1 10sec 0 Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type SystemLiquid Syringe profile 2 Medium 3 Type Reagent Syringe profile 3 Medium 4 Type Destination Syringe profile 6 Medium ID Volumef ul Volumef ul Medium ID Volumef ul Medium ID Volumef ul Assay Details Assay 03 Version 100 Result unit mg L Evaluation Measurement duration 1 10 sec 3600 Evaluation method 8 Two point BNII Blockmode 0 Calibration Calibration method LogitLog Deviation 5 Cal valid until 100 Special parameters Parameter 1 2 Controls Control ID Name Dilution 0776 TBS IgG Sc 1 1 100 0975 TBS IgG Sc 2 1 100 Name G3L Assayclass Sample diluti Transfers 1 Mixing No mixing Buffer 170 Medium ID Air Bubble 2 1302 G2 No Calib long name TBS LigG3 Neph Assaygroup Serum on 1 100 Min dilution 1 5 Measurement transfer 2 Meas window 1 10 sec from 75 until 3600 Overflow 0 Calibrator 1306 IgG Std Start dilution 1 80 Supporting points 5 Deviation 15 15 Start wash 06 goto waste End wash 00 normal Incubation time 1 10sec 0 Incubation delta
44. nding Site Las concentraciones se expresan en mg L 0 2 a os N Media Rango 95 percentil gG1 39 5248 1940 8420 gG2 39 838 225 3000 gG3 39 445 186 853 gG4 39 129 5 784 gG total 39 6640 3270 12700 2 4 anos gG1 36 5495 3150 9450 gG2 36 1146 360 2250 IgG3 36 396 173 676 gG4 36 171 10 537 gG total 36 7770 4680 12500 4 6 a os gG1 49 6149 3060 9450 gG2 49 1496 605 3450 gG3 49 531 99 1221 gG4 49 329 18 1125 IgG total 49 9280 5320 13400 6 8 a os gG1 43 5758 2880 9180 gG2 43 1754 440 3750 gG3 43 437 155 853 gG4 43 331 4 992 gG total 43 8860 4540 13600 8 10 anos gG1 32 6345 4320 10200 gG2 32 2126 720 4300 gG3 32 521 127 853 gG4 32 349 19 932 IgG total 32 9700 5680 13600 10 12 a os gG1 46 6421 4230 10600 gG2 46 1980 760 3550 gG3 46 620 173 1730 gG4 46 323 16 1150 IgG total 46 10200 5680 14900 12 14 anos gG1 54 6581 3420 11500 gG2 54 2486 1000 4550 gG3 54 583 283 1250 gG4 54 409 37 1360 IgG total 54 10500 6640 14900 14 18 a os gG1 48 6057 3150 8550 gG2 48 2614 640 4950 gG3 48 708 230 1960 gG4 48 521 110 1570 IgG total 48 10100 5500 14400 12 Las caracter sticas del ensayo que aqu se muestran se han generado en un Siemens BN II que es un analizador que funciona del mismo modo que el Siemens BN ProSpec 12 1 Precisi n intra ensayo Se ens
45. obes ou contenant des particules de mati re ne doivent pas tre utilis s 8 METHODOLOGIE 8 1 Materiel fourni dans le coffret LK001 P 8 1 1 2 x 1mL BS IgG Sub Std Standard sous classes d IgG 8 1 2 2 x 1mL BS IgG Sub Low Control Contr le bas sous classes d IgG 8 1 3 2 x 1mL BS IgG Sub Control Contr le haut sous classes d IgG 8 1 4 1x 1 6mL Human lgG1 BS Antiserum antis rum pour IgG1 8 1 5 1x 1 6mL Human IgG2 BS Antiserum antis rum pour IgG2 8 1 6 1 x 2 4mL gG3 BS Latex R actif latex pour IgG3 8 1 7 1 x 2 4mL gG4 BS Latex R actif latex pour lgG4 8 1 8 1 x 3mL BS Supplementary Reagent L BS R actif Suppl mentaire L pour IgG3 et IgG4 8 2 References NK006 P NK007 P LK008 P et LK009 P 8 2 1 2x 1mL BS IgG Sub Std Standard sous classes d IgG 8 2 2 2 x 1mL BS IgG Sub Low Control Contr le bas sous classes d IgG 8 2 3 2 x 1mL BS IgG Sub Control Contr le haut sous classes d IgG 8 2 4 4 x 1 6mL antis rum ou 4 x 2 4mL de r actif latex de la sous classe appropri e 8 2 5 2 x 3mL BS Supplementary Reagent L BS R actif Suppl mentaire L dans les coffrets LK008 P et LK009 P uniquement 8 3 Materiel necessaire et non fourni 8 3 1 Mat riel pour le pr l vement et la pr paration des chantillons 8 3 2 Analyseur complet 8 4 Procedure 8 4 1 L utilisateur doit tre famili avec le BN ProSpec avant d utiliser cette proc dure 8 4 2 Pr parer l automate comme indiqu dans le manuel d u
46. ontrols Control ID Name 0776 TBS IgG Sc 1 0975 TBS IgG Sc 2 Name G2 Assayclass Neph Sample dilution 1 20 Overflow 0 long name TBS IgG2 Assaygroup Serum Min dilution 1 5 Measurement transfer 2 Meas window 1 10 sec from 75 until 3600 Calibrator 1306 IgG Std Start dilution 1 5 Supporting points 6 Dilution Deviation 1 20 15 1 20 15 Start wash 06 goto waste Incubation delta 1 10sec 0 Medium ID Buffer Volume ul 170 Medium ID Air Bubble Volume ul 2 Medium ID Volumef ul Medium ID Volume pl Start wash 06 goto waste Incubation delta 1 10sec O Medium ID Buffer Volume pl 170 Medium ID Air Bubble Volume pl 2 Medium ID 1302 G2 Volumef ul Medium ID Volumef ul Name G3L Assayclass Neph Sample dilution 1 100 Overflow 0 End wash 00 normal No Calib End wash 06 park needle No Calib long name TBS LigG3 Assaygroup Serum Min dilution 1 5 Measurement transfer 2 Meas window 1 10 sec from 75 until 3600 Calibrator 1306 IgG Std Start dilution 1 80 Supporting points 5 Dilution Deviation 1 100 15 1 100 15 Start wash 06 goto waste Incubation delta 1 10sec 0 Medium ID Diluens Volume ul 160 Medium ID 1305 SRL Volume ul 10 Medium ID Volumef ul Medium ID Volume pl Start wash 06 goto waste Incubation delta 1 10sec O Medium ID Diluens Volume pl 160
47. pitate or turbidity which is not caused by microbial contamination and does not affect the activity of the antisera In such cases the antiserum should be clarified by filtration through a disposable filter pore size 0 2um or by centrifugation 10mins at approximately 15 000 x g 17 SPECIMEN COLLECTION AND PREPARATION Use fresh or deep frozen serum samples Blood samples should be collected by venepuncture allowed to clot naturally and the serum separated as soon as possible to prevent haemolysis The serum may be stored at 2 8 C for up to 8 days prior to assay or for prolonged storage kept undiluted at 20 C or below Repeated freezing and thawing should be avoided Microbially contaminated haemolysed and lipaemic serum and samples containing particulate matter should not be used 8 METHODOLOGY Materials provided code LK001 P kit 2x 1mL BS IgG Sub Std Human IgG Subclass Standard 2x 1mL BS IgG Sub Low Control Human IgG Subclass Low Control 2x 1mL BS IgG Sub Control Human IgG Subclass High Control 1 x 1 6mL Human IgG1 BS Antiserum Antiserum to IgG1 1x 1 6mL Human lgG2 BS Antiserum Antiserum to lgG2 1 x2 4mL gG3 BS Latex Latex to IgG3 1 x2 4mL gG4 BS Latex Latex to IgG4 1x 3mL BS Supplementary Reagent L for IgG3 and IgG4 assay Code NK006 P NK007 P LK008 P and LK009 P kits 2 x 1mL BS IgG Sub Std Human IgG Subclass Standard 2x 1mL BS IgG Sub Low Control Human IgG Subclass Low Control 2x 1mL BS IgG Sub
48. r Untersuchung eines gesunden Blutspenderkollektivs aus Gro britannien mit diesen Kits und dienen nur zur Orientierung Es wird empfohlen eigene Normalbereiche zu bestimmen 12 2 Inter Assay Variation IgG Subklasse Anzahl Mittelwert Median 95 lle Bereich n mg L mg L mg L IgG1 30 6330 6085 3824 9286 IgG2 30 4528 4541 2418 7003 IgG3 30 790 7 706 4 218 2 1760 6 IgG4 30 280 0 215 3 39 2 864 0 11 2 Kinder Diese Werte wurden mittels Radialer Immundiffusion The Binding Site Single Dilution ermittelt Die Seren der Kinder wurden in einem Krankenhaus von Birmingham England gesammelt Alle Angaben sind in mg L 0 2 Jahre Anzahl Mittelwert 95 ile Bereich gG1 39 5248 1940 8420 gG2 39 838 225 3000 gG3 39 445 186 853 gG4 39 129 5 784 Total IgG 39 6640 3270 12700 2 4 Jahre gG1 36 5495 3150 9450 gG2 36 1146 360 2250 gG3 36 396 173 676 gG4 36 171 10 537 Total IgG 36 7770 4680 12500 4 6 Jahre gG1 49 6149 3060 9450 gG2 49 1496 605 3450 gG3 49 531 99 1221 gG4 49 329 18 1125 Total IgG 49 9280 5320 13400 6 8 Jahre gG1 43 5758 2880 9180 gG2 43 1754 440 3750 gG3 43 437 155 853 gG4 43 331 4 992 Total IgG 43 8860 4540 13600 3 Serum Pr parationen mit unterschiedlichen IgG Subklassen Konzentrationen wurden an mindestens 3 verschiedenen Zeitpunkten bes
49. rective 98 79 EC Annexe Il n anmoins ces tests ne peuvent garantir l absence d agents infectieux Tous les chantillons doivent donc tre manipul s comme des produits potentiellement infectieux Seul un personnel qualifi dans la manipulation d chantillons potentiellement infectieux est autoris utiliser ce coffret ATTENTION Ce produit contient de l azide de sodium et ProClin 300 et doit tre manipul avec pr caution des gants appropri s et d autres v tements de protection doivent tre port s lors de toutes manipulation Ne pas ing rer ou avoir de contact avec la peau sp cialement sur les zones abim es ou des muqueuses S il y a contact laver abondamment avec de l eau et demander un avis m dical Des azides de m taux explosifs peuvent se former par contact prolong entre de Pazide de sodium et les tuyauteries en plomb et cuivre pour liminer les r actifs rincer avec un large volume d eau pour pr venir tout d g t Ce produit ne doit tre utilis que par du personnel entrain Le suivi de ces instructions est essentiel Les r sultats seront consid r s comme invalides si d autres param tres autres que ceux ci sont utilis s Les r actifs de diff rents lots NE SONT PAS interchangeables Si un nombre important de tests est r alis des pr cautions doivent tre prises pour s assurer que les r actifs utilis s sont issus du m me lot 6 STOCKAGE ET STABILITE Les coffrets non ouverts doivent
50. reinander gemischt oder gemeinsam verwendet werden Bei gro em Testdurchsatz muss darauf geachtet werden dass alle Reagenzien der gleichen Charge entstammen 6 LAGERUNG UND STABILIT T Die unge ffneten Kits sind bei 2 8 C bis zum auf dem AuBenetikett angegebenen Verfallsdatum haltbar Unge ffnete Reagenzien Kalibratoren und Kontrollen bei 2 8 C lagern Nach dem ffnen sind sie 4 Wochen bei 2 8 C stabil Reagenzien so lagern dass keine Verdunstung oder Verunreinigung stattfinden kann Offene Flaschen nach Gebrauch nicht im Ger t stehen lassen W hrend der Lagerung kann in den noch unge ffneten Antiseren ein Niederschlag oder eine Trub ng entstehen Dies wird nicht durch mikrobielle Kontamination verursacht und beeinflusst die Aktivit t des Antiserums nicht In solchen F llen das Antiserum durch Filtration durch einen Einweg Filter mit der Porengr e von 0 2 um oder Zentrifugation ca 10 min bei 15000g klaren 7 PROBENENTNAHME UND VORBEREITUNG Immer frisches oder tiefgefrorenes Serum verwenden Blutproben ber Venenpunktur sammeln und auf nat rliche Weise gerinnen lassen Serum so schnell wie m glich vom Gerinnsel trennen um eine H molyse zu vermeiden Die Seren k nnen bei 2 8 C bis zu 8 Tagen vor dem Test gelagert werden F r eine l ngere Lagerung empfiehlt es sich die Seren unverd nnt bei mindestens 20 C einzufrieren Wiederholtes Einfrieren und Auftauen vermeiden Keine mikrobiell oder mit Partikeln verunreinigte
51. ringe profile 7 Medium 2 Type SystemLiquid Syringe profile 2 Medium 3 Type Reagent Syringe profile 3 Medium 4 Type Destination Syringe profile 6 IgG4 Assay Details Assay ID 204 Version 100 Result unit mg L Evaluation Measurement duration 1 10 sec 3600 Evaluation method 8 Two point BNI Blockmode 0 Calibration Calibration method LogitLog Deviation 5 Cal valid until 100 Special parameters Parameter 1 2 Controls Control ID Name 0776 TBS IgG Sc 1 0975 TBS IgG Sc 2 Transfers 1 Mixing No mixing Incubation time 1 10sec O Medium 1 Type SystemLiquid Syringe profile 7 Medium 2 Type Reagent Syringe profile 2 Medium 3 Type Sample Syringe profile 3 Medium 4 Type Destination Syringe profile 3 Start wash 06 goto waste Incubation delta 1 10sec O Medium ID Buffer Volumel ul 170 Medium ID Air Bubble Volume pl 2 Medium ID Volumef ul Medium ID Volumef ul Start wash 06 goto waste Incubation delta 1 10sec O Medium ID Buffer Volumel ul 170 Medium ID Air Bubble Volume ul 2 Medium ID 1302 G2 Volumelul 40 Medium ID Volumelul 112 Name G3L Assayclass Neph Sample dilution 1 100 Overflow 0 End wash 00 normal No Calib End wash 06 park needle No Calib long name TBS LlgG3 Assaygroup Serum Min dilution 1 5 Measurement transfer 2 Meas window 1 10 sec from 75 unti
52. ste kit est preparado para la cuantificaci n de las subclases IgG 1 2 3 y 4 humanas en suero en el analizador BN ProSpec La medida de estas inmunoglobulinas ayuda en el diagn stico del metabolismo anormal de prote nas y la incapacidad del organismo en su resistencia frente a agentes infecciosos 2 RESUMEN Y EXPLICACION La IgG en adultos normales supone aproximadamente el 75 de las inmunoglobulinas s ricas totales Dentro de la clase IgG el orden habitual de concentraci n de las 4 subclases es IgG1 gt lgG2 gt lgG3 gt IgG4 aunque la concentraci n real puede variar mucho entre individuos Las 4 subclases IgG muestran propiedades considerablemente distintas incluyendo la capacidad de fijaci n del complemento uni n a macr fagos y paso a trav s de la placenta Niveles anormales de una o m s subclases pueden estar asociados con ciertas condiciones incluyendo anafilaxis enfermedades autoinmunes e intestinales as como hipo o hiper gammaglobulinemia ref 1 Concretamente una producci n disminuida de lgG2 en ni os puede estar asociada con infecciones recurrentes ref 2 Este tema ha sido revisado refs 3 4 El BN ProSpec permite una automatizaci n completa la cuantificaci n r pida de reacciones de dispersi n de luz para la determinaci n de prote nas 3 PRINCIPIO DEL METODO La evaluaci n de la concentraci n de un antigeno soluble por nefelometria supone la adici n de la muestra a una soluci n que contiene el anti
53. sts sind f r den Gebrauch in der in vitro Diagnostik in der EU freigegeben Directive 98 79 EC Annex Il Es gibt aber zur Zeit keine absolut sicheren Testmethoden zum Auschluss dieser und anderer Infektionstr ger Umgangs und Entsorgungsmethoden sollten denen f r potentiell infekti sem Material entsprechen inklusive dem Tragen entsprechender Schutzkleidung Der Test sollte nur von entsprechend geschultem Personal durchgef hrt werden ACHTUNG Dieses Produkt enth lt Natriumazid und ProClin 300 und muss mit den entsprechenden Vorsichts maBnahmen behandelt werden Das Tragen von Handschuhen und anderer Schutzkleidung wird w hrend des Umgangs mit diesem Produkt empfohlen Verschlucken sowie Kontakt mit Haut v a bei Verletzungen oder Schleimh uten vermeiden Nach Kontakt Hautstelle mit viel Wasser absp len und rztlichen Rat einholen Natriumazid kann mit Blei oder Kupferrohren explosive Metallazide bilden Nach der Entsorgung mit ausreichender Menge Wasser nachsp len um Azidablagerungen zu vermeiden Dieser Test sollte nur f r den angegebenen Verwendungszeck von entsprechend geschultem Laborpersonal durchgef hrt werden Die Einhaltung der Arbeitsanleitung bei allen Arbeitsschritten ist dringend notwendig Bei Verwendung von abge nderten Testparametern kann die Richtigkeit der Ergebnisse nicht garantiert werden Insert Code SINO83 Version 12 August 2010 Page 3 of 10 Reagenzien unterschiedliicher Chargen d rfen NICHT unte
54. tilisation 8 4 3 Les protocoles de tests des sous classes d IgG sont d finir dans l automate 8 4 4 Un ing nieur d application de chez Siemens doit intervenir pour la cr ation de nouveaux protocoles de tests IgG1 Assay Details Assay ID 201 Name G1 Version 100 Assayclass Neph Result unit mg L Sample dilution 1 100 Evaluation Measurement duration 1 10 sec 3600 Measurement transfer 2 Evaluation method 8 Two point BNI Meas window 1 10 sec from 75 until 3600 Blockmode 0 Overflow 0 Calibration Calibration method LogitLog Deviation 5 Cal valid until 100 Special parameters Parameter 1 2 Controls Control ID Name Dilution Deviation 0776 TBS IgG Sc 1 1 100 15 0975 TBS IgG Sc 2 1 100 15 long name TBS IgG1 Assaygroup Serum Min dilution 1 5 Calibrator 1306 IgG Std Start dilution 1 10 Supporting points 6 Transfers 1 Mixing No mixing Start wash 06 goto waste End wash 00 normal Incubation time 1 10sec 0 Incubation delta 1 10sec O Medium 1 Type SystemLiquid Medium ID Buffer Syringe profile 7 Volume pl 170 Medium 2 Type SystemLiquid Medium ID Air Bubble Syringe profile 2 Volume pl 2 Medium 3 Type Sample Medium ID No Calib Syringe profile 3 Volumef ul Medium 4 Type Destination Medium ID Syringe profile 3 Volumef ul Transfers 2 Mixing normal Start wash 06 goto waste End wash 06 park needle Incubation time 1 1
55. timmt Alle Konzentrationsangaben sind in mg L N Anzahl der Zeitpunkte IgG1 IgG2 IgG3 IgG4 z VK A VK A VK p NK Mittelwert Mittelwert Mittelwert Mittelwert N N N N 4 38 6 84 4 41 4 10 Serum 1 40765 6 7811 6 697 5 3 532 6 3 2 39 4 18 2 38 4 13 Serum 2 12402 6 1621 6 525 9 a 387 7 3 6 95 2 43 3 00 4 38 Serum 3 3043 780 1 353 1 319 6 6 6 3 4 BN ist ein eingetragenes Warenzeichen von Siemens Healthcare Diagnostics Inc CA REFERENZEN 1 Shakib F amp Stanworth D R 1980 Human IgG subclasses in health and disease Part 11 La Ricerca Clin Lab 10 561 580 2 Shackleford P G Polmar S H et al 1986 Spectrum of IgG2 subclass deficiency in children with recurrent infections Prospective study J Pediatr 108 647 653 3 Hamilton R H 1987 Human IgG subclass measurements in the clinical laboratory Clin Chem 33 1707 1725 4 Schur P H 1987 IgG subclasses a review Annals of Allergy 58 89 99 5 Carr Smith H D Overton J Bradwell A R 1997 IgG subclass value assignment to the protein reference preparation CRM470 Clin Chem Vol 43 No 56 PS238 Insert Code SIN083 Version 12 August 2010 Page 5 of 10 REACTIFS LIQUIDES POUR LE DOSAGE DES SOUS CLASSES D IgG HUMAINES pour une utilisation sur automate Siemens BN ProSpec Pour un usage en diagnostic in vitro uniquement Reference LK001 P NK006 P NK007 P LK008 P LK009 P Produit fabr
56. trol values fall outside 15 limits against new calibration curves check the instument before repeating the assay If problems persist refer to the supplier 10 4 The results obtained from measuring IgG subclasses should not be used in assessing atopy in allergic patients 10 5 Diagnosis cannot be made and treatment must not be initiated on the basis of IgG subclass measurements alone Clinical history and other laboratory findings must also be taken into account 11 EXPECTED VALUES The ranges provided below have been obtained from a limited number of British samples and are intended for guidance purposes only Wherever possible it is strongly recommended that local ranges are generated 11 1 Adult normal ranges These ranges were obtained by measuring the subclass content of sera provided by the Birmingham Blood Transfusion Service taken from healthy adult donors IgG Number Mean Median 95 Percentile Subclass n mg L mg L Range mg L IgG1 30 6330 6085 3824 9286 IgG2 30 4528 4541 2418 7003 IgG3 30 790 7 706 4 218 2 1760 6 IgG4 30 280 0 215 3 39 2 864 0 11 2 Paediatric ranges These ranges were obtained by measuring the Total IgG and subclass content of paediatric serum samples from a Birmingham Hospital using Binding Site Radial Immunodiffusion products All concentrations are in mg L 0 2 years N Mean 95 ile Range gG1 39 5248 19
57. vante 4 3 Calibradores y controles Preparados a partir de una mezcla estabilizada de suero humano se suministran en forma l quida estabilizada Conservantes azida s dica 0 099 EACA 0 1 y benzaminida 0 01 La concentraci n de las subclases IgG indicada en la etiqueta se ha obtenido por comparaci n con el material de referencia CRM470 ref 5 4 4 Reactivo suplementario Contiene azida s dica 0 099 como conservante ProClin es una marca registrada de Rohm and Haas Corp Philadelphia PA Los sueros humanos suministrados en el kit han sido sometidos a screening para donantes resultando negativos a la presencia del ant geno de superficie de la hepatitis B y a la presencia de los anticuerpos de la ante los virus HIV1 HIV2 y HCV Las t cnicas usadas est n aprobadas por la FDA USA o para el diagn stico in vitro por la UE Directiva 98 79 EC Anexo Il Sin embargo los sobredichos ensayos no garantizan la ausencia de agentes infecciosos Por lo tanto deben tratarse los reactivos como potencialmente infecciosos Tanto la manipulaci n como los m todos de eliminaci n de desechos deber n realizarse conforme a la normativa de materiales infecciosos y solo personal adecuadamente instruido deber efectuar el test AVISO Este kit contiene azida s dica y ProClin 300 y debe manipularse con precauci n use guantes y vestuario protector adecuado en todo momento al manipular este producto No trague ni permita el contacto con la pie
58. vo l tex de la subclase correspondiente 8 2 5 2 x 3 0mL BS Supplementary Reagent L i BS Reactivo suplementario L para los ensayos LK008 P y LK009 P 8 3 Materiales necesarios pero no suministrados 8 3 1 Materiales necesarios para la recolecci n y preparaci n de las muestras 8 3 2 Un analizador funcional y equipado 8 4 Procedimiento de ensayo 8 4 1 El usuario debe estar familiarizado con el manejo del BN ProSpec antes de llevar a cabo procedimientos de ensayo 8 4 2 Prepare el analizador seg n el manual de usuario 8 4 3 Los protocolos de ensayo de subclases IgG subclass assay est n definidos en el analizador 8 4 4 Es necesario un representante de Siemens para crear nuevos protocolos de ensayo IgG1 Assay Details Assay ID 201 Version 100 Result unit mg L Evaluation Measurement duration 1 10 sec 3600 Evaluation method 8 Two point BNI Blockmode 0 Calibration Calibration method LogitLog Deviation 5 Cal valid until 100 Special parameters Parameter 1 2 Controls Control ID Name Dilution Deviation Level 0776 TBS IgG Sc 1 1 100 15 Medium 0975 TBS IgG Sc 2 1 100 15 Low Name G1 Assayclass Neph Sample dilution 1 100 long name TBS IgG1 Assaygroup Serum Min dilution 1 5 Measurement transfer 2 Meas window 1 10 sec from 75 Overflow 0 until 3600 Calibrator 1306 IgG Std Start dilution 1 10 Supporting points 6 Transfers 1 Mixing No mixing Start wash 06
59. well A R 1997 IgG subclass value assignment to the protein reference preparation CRM470 Clin Chem Vol 43 No 56 PS238 Nur zur in vitro Diagnostik Bestell Nr LK001 P NK006 P NK007 P LK008 P LK009 P Hergestellt von The Binding Site Group Ltd PO Box 11712 Birmingham B14 4ZB UK www bindingsite co uk Vertrieb in Deutschland und sterreich durch The Binding Site GmbH Robert Bosch Stra e 2A D 68723 Schwetzingen Deutschland Telefon 49 0 6202 92 620 Fax 49 0 6202 92 62 222 e mail office bindingsite de BN ProSpec ist ein eingetragenes Warenzeichen von Siemens Healthcare Diagnostics Inc Fi VERWENDUNGSZWECK Zur quantitativen Bestimmung von Human IgG Subklassen 1 2 3 und 4 im Serum auf dem BN ProSpec Die Bestimmung dieser Immunglobuline dient zur Unterst tzung der Diagnose eines abnormalen Proteinmetabolismus und das Unverm gen des K rpers auf infekti se Erreger zu reagieren 2 EINLEITUNG Bei gesunden Erwachsenen stellt IgG ca 75 des Gesamtimmunglobulins dar Innerhalb der IgG Klasse nimmt die Konzentration in der Reihenfolge IgG1 gt IgG2 gt IgG3 gt IgG4 ab wobei der tats chliche Gehalt individuell deutlich variieren kann Die vier IgG Subklassen zeigen deutliche Unterschiede in ihren Eigenschaften wie z B F higkeit zur Komplementbindung Makrophagenbindung und Placentadurchl ssigkeit Von der Norm ab weichende Werte einer oder mehrerer Subklassen k nnen mit verschiedenen
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