WHO Prequalification of In Vitro Diagnostics Programme PUBLIC REPORT Product: Enzygnost HBsAg 6.0 WHO reference number: PQDx 0173-064-00 Enzygnost HBsAg 6.0 with product codes OPFM03, OPFM05, OPFM07(Q) and Supplementary reagents kit for Enzygnost /TMB with product code OUVP17 manufactured by Siemens Healthcare Diagnostics Products GmbH, CE-mark regulatory version, was accepted for the WHO list of prequalified in vitro diagnostics and was listed on 22 March 2016. This report was amended on 04 April 2016 to correct a typographical error. Intended use: Enzygnost HBsAg 6.0 is an enzyme immunoassay for the qualitative detection of hepatitis B (surface) antigen in human serum or plasma. The enzyme immunoassay can be processed using the ELISA processors, BEP III System, BEP 2000 System or BEP 2000 Advance System, as well as the Quadriga and Quadriga BeFree Systems. A nonautomated processing of the test procedure is also possible. Assay description: Enzygnost HBsAg 6.0 is an enzyme immunoassay based on a two-step method. In the first step, the HBsAg contained in the specimen reacts simultaneously with the polyclonal anti- HBs antibodies attached to the wells of the microtitration plates and with Conjugate 1 (Anti-HBs/Biotin, colored blue). In the second step, after removing the unbound reactants, Conjugate 2 (Streptavidin/POD, colored yellow) then reacts with Conjugate 1. After removing the unbound reactants, the enzyme activity of the bound Conjugate 2 is determined (blue color reaction). The enzymatic conversion of the chromogen is terminated by the addition of Stopping Solution POD (yellow color reaction). The color intensity is proportional to the concentration of antigen in the specimen. Test kit contents: Component Test plate Microtitration plate coated with sheep antibodies to HBsAg Conjugate 1 Anti-HBs/Biotin Monoclonal anti-hbs (mouse), biotin-conjugated, coloured yellow (Contains preservative phenol ( 1 2 x 96 tests/kit Product code OPFM03 10 x 96 tests/kit Product code OPFM05 10x96 tests/kit Product code OPFM07(Q) 2 x 96 wells 10 x 96 wells 10x 96 wells 2 bottles x 5 ml 10 bottles x 5 ml 3 bottles x 15 ml Page 1 of 32
Component g/l) 2 x 96 tests/kit Product code OPFM03 10 x 96 tests/kit Product code OPFM05 10x96 tests/kit Product code OPFM07(Q) Conjugate 2 Streptavidin/POD 2 x 12.5 ml 10 x 12.5 ml 2 x 75 ml Streptavidin, peroxidase (POS)- conjugated, coloured yellow (Contains preservative phenol ( 1 g/l) HBsAg Control Serum, negative 2 x 2.5 ml 4 x 2.5 ml 3 x 2.5 ml Stabilized human serum, coloured red (Contains preservative phenol ( 1 g/l) HBsAg Control Serum, positive 2 x 1.5 ml 3 x 1.5 ml 3 x 1.5 ml Stabilized human serum, 0.15 to 0.32 U/ml (target value 0.25 U/ml), coloured green (Contains preservative phenol ( 1 g/l) Label "Empty bottle for conjugate N/A 1 unit NA 1" Label "Empty bottle for conjugate N/A 1 unit NA 2" Instructions for use 1 unit 1 unit 1 unit Polyethylene bag 1 unit 1 unit 1 unit Items required but not provided: Item Product code Supplementary reagents kit for OUVP17 Enzygnost /TMB Buffer/Substrate TMB Chromogen TMB Stopping Solution POD Washing Solution POD Adhesive foils Empty bottle for the Chromogen Working Solution Instructions for Use Non-automated processing instrumentation requirements Page 2 of 32
Incubator N/A Microtitration plate washer N/A Spectrophotometer suitable for 96-well plates N/A (450nm measuring and 650nm reference wavelengths) Automated processing instrumentation requirements BEP III System: for automated processing and TBC evaluation of the test after manual dispensing of samples and controls BEP 2000 System/BEP 2000 Advance System: TBC for fully automated processing and evaluation of the test Quadriga System: for fully automated TBC processing and evaluation of the test in combination with BEP III Precision pipettes plus tips N/A Storage: The test kit should be stored at 2-8 C. Shelf-life: 12 months. Warnings/Limitations: See manufacturer s instructions for use. WHO special warnings: WHO reviewed the instructions for use that were current at the time of WHO prequalification, and a number of changes were suggested. Most changes, but not all, were made by the manufacturer (WHO comments relate to intended use, warnings and precautions, preparation of reagents, certain aspects of test procedure, results, nomenclature). Page 3 of 32
Summary of prequalification assessment for Enzygnost HBsAg 6.0 Date Outcome Amended PQ 04 April 2016 Status on PQ list 22 March 2016 listed Dossier assessment N/A MR Inspection status 13 April 2015 MR Laboratory evaluation 01 December 2015 MR MR: Meets Requirements N/A: Not Applicable Prioritization for prequalification Based on the established criteria, Enzygnost HBsAg 6.0 was given priority for WHO prequalification. Product dossier assessment In accordance with the WHO procedure for abbreviated prequalification assessment, Siemens Healthcare Diagnostics Products GmbH was not required to submit a product dossier for Enzygnost HBsAg 6.0 as per the Instructions for compilation of a product dossier (PQDx_018 v1). Notwithstanding, certain aspects of the product dossier previously submitted for stringent regulatory review were reviewed by an assessor during the site inspection. Manufacturing site inspection In accordance with the WHO procedure for abbreviated prequalification assessment, a shortened inspection with fewer inspectors was conducted at the site of manufacture (Emil-von-Behring Straße 76, 35041 Marburg, Germany) of Enzygnost HBsAg 6.0 in 24-26 February, 2015 as per the Information for manufacturers on prequalification inspection procedures for the sites of manufacture of diagnostics (PQDx_014 v1). The inspection found that the manufacturer had an acceptable quality management system and good manufacturing practices in place that ensured the consistent manufacture of a product of good quality. The inspection was concluded with the final inspection report sent 13 April 2015. Based on the site inspection and corrective action plan review, the quality management system for Enzygnost HBsAg 6.0 meets WHO prequalification requirements. Page 4 of 32
Laboratory evaluation Enzygnost HBsAg 6.0 was evaluated by WHO in the 3rd quarter of 2015 using plasma specimens. From this evaluation, we drew the following conclusions: Enzygnost HBsAg 6.0 is an enzyme immunoassay for the detection of HBsAg in human serum or plasma specimens. A volume of 100µl of specimen is needed to perform the assay. This type of assay requires laboratory equipment and cannot be performed in laboratories with limited facilities. Reading of the results must be performed with a spectrophotometer. In this limited performance evaluation on a panel of 513 specimens, we found an initial sensitivity (95% CI) of 100.0% (98.2 100.0%) and an initial specificity (95% CI) of 99.4% (97.7 99.99%) compared to the reference results. The final sensitivity (95% CI) was 100.0% (98.2 100.0%) and the final specificity (95% CI) was 100% (98.8 100.0%) compared to the reference results. In this study, 0% of the results were recorded as indeterminate. Lot to lot variation was in the acceptance range. For six seroconversion panels Enzygnost HBsAg 6.0 assay detected HBsAg seroconversion on average 0.5 specimens earlier than the benchmark assay (Monolisa Ag HBs Plus [Bio- Rad Laboratories]). For the low titer panel, Enzygnost HBsAg 6.0 correctly classified all specimens. For the 1st International Biological Reference Preparation for Hepatitis B surface antigen [NIBS code 5086/08], Enzygnost HBsAg 6.0 detected the 0.13IU/mL. Page 5 of 32
Labelling 1. Labels 2. Instructions for use Page 6 of 32
1. Labels 1.1 Overview on kit components Product codes of variants OPFM 03 (2 x 96) Overview on kit components OPFM 05 (10 x 96) OPFM 07 (10 x 96 (Q)) Page 7 of 32
1.2 Table of assigned values (TAV) Component TAV Document Example: Page 8 of 32
1.3 Labels for product variant OPFM 03 (2x96) Component Outer Box Label MTP MTP pouch with label MTP side label (EXAMPLE) Page 9 of 32
Component Conjugate 1 Label Conjugate 2 Page 10 of 32
Component Control - Label Control + Page 11 of 32
1.4 Labels for product variant OPFM 05 (10x96) Component Outer Box Label MTP MTP pouch with label MTP side label (EXAMPLE) Page 12 of 32
Component Conjugate 1 Label Conjugate 1 empty vial Page 13 of 32
Component Conjugate 2 Label Conjugate 2 empty vial Page 14 of 32
Component Control - Label Control + Page 15 of 32
1.5 Labels for product variant OPFM 13 (10x96 Q) Component Outer box Label MTP MTP pouch with label MTP side label (EXAMPLE) Page 16 of 32
Component Conjugate 1 Label Conjugate 2 Page 17 of 32
Component Control - Label Control + Page 18 of 32
2. Instructions for use (excerpt containing only the English version) Page 19 of 32
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