WHO Prequalification of Diagnostics Programme PUBLIC REPORT Product: Bioelisa HCV 4.0 Number: PQDx 0165-060-00 Abstract Bioelisa HCV 4.0 with product codes 3000-1115 and 3000-1116, manufactured by Biokit S.A., CE-mark regulatory version, was accepted for the WHO list of prequalified diagnostics and was listed on 31 March 2015, and amended on 8 April 2015 and on 13 November 2015. Bioelisa HCV 4.0 is an enzyme immunoassay in which the wells of a microplate are coated with recombinant antigens representing epitopes of HCV: Core, NS3, NS4 and NS5. Serum or plasma specimens are added to these wells. If antibodies specific for HCV are present in the specimen, they will form stable complexes with the HCV antigens on the well. Excess specimen is removed by a wash step and a rabbit anti-human IgG conjugated with peroxidase is then added and allowed to incubate. The conjugate will bind to any antigenantibody complexes formed. After a second wash, a solution of enzyme substrate and chromogen is added. This solution will develop a blue colour if the specimen is reactive. The blue colour changes to yellow after blocking the reaction with sulphuric acid. The intensity of colour is proportional to anti-hcv antibodies concentration in the specimens. Wells containing negative specimens remain colourless. All initially reactive specimens should be retested in duplicate to confirm the initial result. The test kit contains: 96 well plate 12 x 8 wells coated with HCV recombinant antigens (E. coli). Individually separable wells. Concentrate conjugate Rabbit anti-human IgG antibodies conjugated with peroxidase. Contains red dye, stabilisers protein, 0.02% thimerosal and 0.001% gentamicin sulphate. To be diluted 1/51 with the conjugate diluent before use. Conjugate diluent Tris buffer containing yellow dye, additives, 0.02% 96 tests Product code 3000-1115 1x 96 well plate 1x 0.40 ml 1x 15 ml 480 tests Product code 3000-1116 5x 96 well plates 1x 1.5 ml 1x 70 ml Page 1 of 19
thimerosal and 0.001% gentamicin sulphate Sample diluent 1x 30 ml 1x 120 ml Tris buffer with stabilisers protein, < 0.1% sodium azide, < 1.0% Triton X-100 as preservative and 8% Ethylene Glycol. Ready to use. Concentrated washing solution 3x 50 ml 5x 100 ml Phosphate buffer concentrate containing 1% Tween 20 and 0.01% thimerosal. To be diluted 1/10 in distilled or deionised water before use. Substrate buffer 1x 14 ml 5x 14 ml Citrate-acetate buffer containing hydrogen peroxide and 0.002% gentamicin sulphate. Chromogen 1x 1.5 ml 1x 1.5 ml 3,3', 5,5'-Tetramethylbenzidine (TMB) dissolved in dimethylsulphoxide (DMSO) High positive control 1 x 3 ml 1x 5 ml Diluted human serum containing antibodies to HCV. Contains < 0.1% sodium azide, < 1.0% Triton X-100 and green dye. Ready to use. Low positive control 1 x 7 ml 1x 7 ml Diluted human serum containing antibodies to HCV. Contains < 0.1% sodium azide, < 1.0% Triton X-100 and green dye. Ready to use. Negative control 1x 5 ml 1x 5 ml Diluted human serum negative for antibodies to HCV. Contains < 0.1% sodium azide, < 1.0% Triton X-100 and yellow dye. Ready to use. Stopping solution 1x 12 ml Not included 1N sulphuric acid. Ready to use. Resealable bag 1 1 adhesive sealers 6 20 Storage: The test kit should be stored at 2 to 8 C. Shelf-life: 12 months. Page 2 of 19
Summary of prequalification status for Bioelisa HCV 4.0 Initial acceptance Date Outcome Amended PQ 13 November 2015 listed Status on PQ list 31 March 2015 listed Dossier assessment 21 August 2014 MR Inspection status 18 April 2014 MR Laboratory evaluation 27 March 2015 MR MR: Meets Requirements NA: Not Applicable Bioelisa HCV 4.0 was accepted for the WHO list of prequalified diagnostics on the basis of data submitted and publicly available information. This public report was amended 13 November 2015 to reflect that the outstanding commitment to prequalification had been fulfilled. Background information Biokit S.A. submitted an application for prequalification of Bioelisa HCV 4.0. Based on the established prioritization criteria, Bioelisa HCV 4.0 was given priority for prequalification. Product dossier assessment Biokit S.A. submitted a product dossier for Bioelisa HCV 4.0 as per the Instructions for compilation of a product dossier (PQDx_018 v1). The information submitted in the product dossier was reviewed by WHO staff and external experts (assessors) appointed by WHO in accordance with the internal report on the screening and assessment of a product dossier (PQDx_009 v2). Based on the product dossier screening and assessment findings, a recommendation was made to accept the product dossier for Bioelisa HCV 4.0 for prequalification. Manufacturing site inspection A comprehensive inspection was performed at the site of manufacture (Can Malé, 08186 Llicà d`amunt, Barcelona, Spain) of Bioelisa HCV 4.0 in March 2014 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 a well implemented quality management system and good manufacturing practices in place that ensured the consistent manufacture of a product of good quality. The manufacturer's Page 3 of 19
responses to the nonconformities found at the time of the inspection were accepted 18 April 2014. Laboratory evaluation Bioelisa HCV 4.0 (Biokit S.A.) was evaluated by WHO in the 4th quarter of 2014 using plasma specimens. From this evaluation, we drew the following conclusions: Bioelisa HCV 4.0 is an enzyme immunoassay for the detection of HCV antibodies in human serum/plasma. A volume of 10µl 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 spectrophotometric EIA reader. In this limited performance evaluation on a panel of 466 specimens, we found an initial sensitivity (95% CI) of 100% (97.6 100%) and an initial specificity (95% CI) of 99.0% (97.2 99.8%) compared to the reference results. The final sensitivity (95% CI) was 100% (97.6 100%) and the final specificity (95% CI) was 100% (98.8 100%) compared to the reference results. Lot to lot variation was in the acceptable range for nine of the ten diluted panels. For four seroconversion panels, Bioelisa HCV 4.0 detected infection on average 1.25 specimens later than the benchmark assay (Ortho HCV 3.0 Enhanced SAVe [Ortho Clinical Diagnostics]). For the worldwide panels, Bioelisa HCV 4.0 correctly classified all specimens. For the two low titer panels Bioelisa HCV 4.0 correctly identified 14 of 28 HCV seropositive specimens. One test run was invalid based on the run validation criteria outlined in the instructions for use, and these results were discarded. On this particular plate, the run failed to meet the manufacturer s validation criteria for the positive control. Page 4 of 19
Labelling 1. Labels 2. Instructions for use Page 5 of 19
1. Labels For product code 3000-1115 (96 tests/kit) Page 6 of 19
For product code 3000-1116 (480 tests/kit) Page 7 of 19
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2. Instructions for use Page 10 of 19
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