# Commission Decision of 7 May 2002 on common technical specifications for in vitro-diagnostic medical devices (notified under document number C(2002) 1344) (Text with EEA relevance) (2002/364/EC), Division 3: COMMON TECHNICAL SPECIFICATIONS (CTS) FOR PRODUCTS REFERRED TO IN ANNEX II, LIST A OF DIRECTIVE 98/79/EC Extent: E+W+S+N.I. Table 1 Screening assays: anti-HIV 1/2, HIV 1/2 Ag/Ab, anti-HTLV I/II, anti-HCV, HCV Ag/Ab, HBsAg, anti-HBc anti-HIV 1/2, HIV 1/2 Ag/Ab Anti-HTLV-I/II anti-HCV, HCV Ag/Ab HBsAg Anti-HBc Diagnostic sensitivity Positive specimens 400 HIV-1 100 HIV-2 including 40 non-B-subtypes, all available HIV/1 subtypes shall be represented by at least 3 samples per subtype 300 HTLV-I 100 HTLV-II 400 (positive samples) Including samples from different stages of infection and reflecting different antibody patterns. Genotype 1-4: > 20 samples per genotype (including non-a subtypes of genotype 4); 5: > 5 samples; 6: if available 400 including subtype-consideration 400 including evaluation of other HBV-markers Sero-conversion panels 20 panels 10 further panels (at Approved Body or manufacturer) To be defined when available 20 panels 10 further panels (at Approved Body or manufacturer) 20 panels 10 further panels (at Approved Body or manufacturer) To be defined when available Analytical sensitivity Standards 0,130 IU/ml (WHO International Standard: Third International Standard for HBsAg, subtypes ayw1/adw2, HBV genotype B4, NIBSC code: 12/226) Specificity Unselected donors (including first-time donors) 5 000 5 000 5 000 5 000 5 000 Hospitalized patients 200 200 200 200 200 Potentially cross-reacting blood-specimens (RF+, related viruses, pregnant women, etc.) 100 100 100 100 100 Table 2 NAT assays for HIV1, HCV, HBV, HTLV I/II (qualitative and quantitative; not molecular typing) European Pharmacopoeia guideline. Notes: Acceptance criteria for ‘whole system failure rate leading to false-neg results’ is 99/100 assays positive. For quantitative NATs a study shall be performed on at least 100 positive specimens reflecting the routine conditions of users (e.g. no pre-selection of specimens). Comparative results with another NAT test system shall be generated in parallel. For qualitative NATs a study on diagnostic sensitivity shall be performed using at least 10 sero-conversion panels. Comparative results with another NAT test system shall be generated in parallel. HIV1 HCV HBV HTLV I/II Acceptance criteria NAT qualitative quantitative qualitative quantitative qualitative quantitative qualitative quantitative As for HIV quantitative As for HIV quantitative As for HIV quantitative Sensitivity Detection limit Detection of analytical sensitivity (IU/ml; defined on WHO standards or calibrated reference materials) According to EP validation guideline : several dilution series into borderline concentration; statistical analysis (e.g. Probit analysis) on the basis of at least 24 replicates; calculation of 95 % cut-off value Detection limit: as for qualitative tests; Quantification limit: dilutions (half-log10 or less) of calibrated reference preparations, definition of lower, upper quantification limit, precision, accuracy, ‘ linear ’ measuring range, ‘ dynamic range ’ . Reproducibility at different concentration levels to be shown According to EP validation guideline : several dilution series into borderline concentration; statistical analysis (e.g. Probit analysis) on the basis of at least 24 replicates; calculation of 95 % cut-off value According to EP validation guideline : several dilution series into borderline concentration; statistical analysis (e.g. Probit analysis) on the basis of at least 24 replicates; calculation of 95 % cut-off value According to EP validation guideline : several dilution series into borderline concentration; statistical analysis (e.g. Probit analysis) on the basis of at least 24 replicates; calculation of 95 % cut-off value Genotype/subtype detection/quantification efficiency At least 10 samples per subtype (as far as available) Dilution series of all relevant genotypes/subtypes, preferably of reference materials, as far as available At least 10 samples per genotype (as far as available) As far as calibrated genotype reference materials are available As far as calibrated genotype reference materials are available Cell culture supernatants (could substitute for rare HIV-1 subtypes) Transcripts or plasmids quantified by appropriate methods may be used. According to EP validation guideline as far as calibrated subtype reference materials are available; in vitro transcripts could be an option According to EP validation guideline as far as calibrated subtype reference materials are available; in vitro transcripts could be an option According to EP validation guideline as far as calibrated subtype reference materials are available; in vitro transcripts could be an option According to EP validation guideline as far as calibrated subtype reference materials are available; in vitro transcripts could be an option Diagnostic specificity negative samples 500 blood donors 100 blood donors 500 blood donors 500 blood donors 500 individual blood donations Potential cross-reactive markers By suitable assay design evidence (e.g. sequence comparison) and/or testing of at least 10 human retrovirus (e.g. HTLV)-positive samples As for qualitative tests By assays design and/or testing of at least 10 human flavivirus (e.g. HGV, YFV) positive samples By assays design and/or testing of at least 10 other DNA-virus positive samples By assay design and/or testing of at least 10 human retrovirus (e.g. HIV-) positive samples Robustness As for qualitative tests Cross-contamination At least 5 runs using alternating high positive (known to occur naturally) and negative samples At least 5 runs using alternating high positive (known to occur naturally) and negative samples At least 5 runs using alternating high positive (known to occur naturally) and negative samples At least 5 runs using alternating high positive (known to occur naturally) and negative samples Inhibition Internal control preferably to go through the whole NAT procedure Internal control preferably to go through the whole NAT procedure Internal control preferably to go through the whole NAT procedure Internal control preferably to go through the whole NAT procedure Whole system failure rate leading to false-neg results At least 100 samples virus-spiked with 3 × the 95 % pos cut-off concentration At least 100 samples virus-spiked with 3 × the 95 % pos cut-off concentration At least 100 samples virus-spiked with 3 × the 95 % pos cut-off concentration At least 100 samples virus-spiked with 3 × the 95 % pos cut-off concentration 99/100 assays positive Table 3 Rapid tests: anti-HIV 1 and 2, anti-HCV, HBsAg, anti-HBc, anti-HTLV I and II Anti-HIV 1/2 Anti-HCV HBsAg Anti-HBc Anti-HTLV I/II Acceptance criteria Diagnostic sensitivity Positive specimens Same criteria as for screening assays Same criteria as for screening assays Same criteria as for screening assays Same criteria as for screening assays Same criteria as for screening assays Same criteria as for screening assays Sero-conversion panels Same criteria as for screening assays Same criteria as for screening assays Same criteria as for screening assays Same criteria as for screening assays Same criteria as for screening assays Same criteria as for screening assays Diagnostic specificity Negative specimens 1 000 blood donations 1 000 blood donations 1 000 blood donations 1 000 blood donations 1 000 blood donations ≥ 99 % (anti-HBc: ≥ 96 %) 200 clinical specimens 200 clinical specimens 200 clinical specimens 200 clinical specimens 200 clinical specimens 200 samples from pregnant women 200 samples from pregnant women 200 samples from pregnant women 200 samples from pregnant women 100 potentially interfering samples 100 potentially interfering samples 100 potentially interfering samples 100 potentially interfering samples 100 potentially interfering samples Table 4 Confirmatory/supplementary assays for anti-HIV 1 and 2, anti-HTLV I and II, anti-HCV, HBsAg Acceptance criteria no neutralisation for HBsAg confirmatory assay. Anti-HIV confirmatory assay Anti-HTLV confirmatory assay HCV supplementary assay HBsAg confirmatory assay Acceptance criteria Diagnostic sensitivity Positive specimens 200 HIV-1 and 100 HIV-2 200 HTLV-I and 100 HTLV-II 300 HCV (positive samples) 300 HBsAg Correct identification as positive (or indeterminate), not negative Including samples from different stages of infection and reflecting different antibody patterns Including samples from different stages of infection and reflecting different antibody patterns. Genotypes 1 – 4: > 20 samples (including non-a subtypes of genotype 4); 5: > 5 samples; 6: if available Including samples from different stages of infection 20 ‘high pos’ samples (> 26 IU/ml); 20 samples in the cut-off range Sero-conversion panels 15 sero-conversion panels/low titre panels 15 sero-conversion panels/low titre panels 15 sero-conversion panels/low titre panels Analytical sensitivity Standards Second International Standard for HBsAg, subtype adw2, genotype A, NIBSC code: 00/588 Diagnostic specificity Negative specimens 200 blood donations 200 blood donation 200 blood donations 10 false positives as available from the performance evaluation of the screening assay . No false-positive results/ no neutralisation 200 clinical samples including pregnant women 200 clinical samples including pregnant women 200 clinical samples including pregnant women 50 potentially interfering samples, including samples with indeterminate results in other confirmatory assays 50 potentially interfering samples including samples with indeterminate results in other confirmatory assays 50 potentially interfering samples including samples with indeterminate results in other supplementary assays 50 potentially interfering samples Table 5 HIV 1 antigen, HIV Ag/Ab, HCV antigen, HCV Ag/Ab The total number of seroconversion panels for combined Ag/Ab assays (from tables 1 and 5) need not be greater than 30. HIV-1 antigen and HIV Ag/Ab assays HCV antigen and HCV Ag/Ab assays Acceptance criteria Diagnostic sensitivity Positive specimens 50 HIV-1 antigen positive 50 cell culture supernatants including different HIV-1 subtypes and HIV-2 25 HCV core antigen and/or HCV RNA positive but anti-HCV negative samples, comprising HCV genotypes 1-6 (if a genotype is not available, a justification shall be made) See general principle in § 3.1.8 Sero-conversion panels 20 sero-conversion panels/low titre panels 20 sero-conversion panels/low titre panels Analytical sensitivity Standards HIV-1 p24 Antigen, First International Reference Reagent, NIBSC code: 90/636 HCV core antigen detection limit shall be investigated using dilutions of the WHO International HCV core antigen Standard: (HCV core Ag product code: PEI 129096/12) For HIV-1 p24 antigen: ≤ 2 IU/ml Diagnostic specificity 200 blood donations 200 clinical samples 50 potentially interfering samples 200 blood donations, 200 clinical samples, 50 potentially interfering samples ≥ 99,5 % after neutralisation or, if no neutralisation test available, after resolution of the sample status according to general principles in § 3.1.5 Table 6 Serotyping and genotyping assay: HCV HCV serotyping and genotyping assay Acceptance criteria Diagnostic sensitivity Positive specimens 200 (positive samples) Including samples from different stages of infection and reflecting different antibody patterns. Genotypes 1 – 4: > 20 samples (including non-a subtypes of genotype 4); 5: > 5 samples; 6: if available ≥ 95 % agreement between serotyping and genotyping > 95 % agreement between genotyping and sequencing Diagnostic specificity Negative specimens 100 Table 7 HBV markers: anti-HBs, anti HBc IgM, anti-HBe, HBeAg Anti-HBs Anti-HBc IgM Anti-HBe HBeAg Acceptance criteria Diagnostic sensitivity Positive specimens 100 vaccinees 200 200 200 ≥ 98 % 100 naturally infected persons Including samples from different stages of infection (acute/chronic, etc.) The acceptance criteria should only be applied on samples from acute infection stage. Including samples from different stages of infection (acute/chronic, etc.) Including samples from different stages of infection (acute/chronic, etc.) Sero-conversion panels 10 follow-ups or anti-HBs sero-conversions When available Analytical sensitivity Standards WHO First International Reference Preparation 1977; NIBSC, United Kingdom HBe — Referenzantigen 82; PEI Germany Anti-HBs: < 10 mIU/ml Diagnostic specificity Negative specimens 500 200 blood donations 200 blood donation 200 blood donations ≥ 98 % Including clinical samples 200 clinical samples 200 clinical samples 200 clinical samples 50 potentially interfering samples 50 potentially interfering samples 50 potentially interfering samples 50 potentially interfering samples Table 8 HDV markers: anti-HDV, anti-HDV IgM, delta antigen Anti-HDV Anti-HDV IgM Delta antigen Acceptance criteria Diagnostic sensitivity Positive specimens 100 50 10 ≥ 98 % Specifying HBV markers Specifying HBV markers Specifying HBV markers Diagnostic specificity Negative specimens 200 200 200 ≥ 98 % Including clinical samples Including clinical samples Including clinical samples 50 potentially interfering samples 50 potentially interfering samples 50 potentially interfering samples Table 9 Blood group antigens in the ABO, Rh and Kell blood group systems 1 2 3 Specificity Number of tests per recommended method Total number of samples to be tested for a launch product Total number of samples to be tested for a new formulation, or use of well-characterised reagents Anti-ABO1 (anti-A), anti-ABO2 (anti-B), anti-ABO3 (anti-A,B) 500 3 000 1 000 Anti-RH1 (anti-D) 500 3 000 1 000 Anti-RH2 (anti-C), anti-RH4 (anti-c), anti-RH3 (anti-E) 100 1 000 200 Anti-RH5 (anti-e) 100 500 200 Anti-KEL1 (anti-K) 100 500 200 Table 11 Variant Creutzfeldt-Jakob disease (vCJD) assays for blood screening Material Number of specimens Acceptance Criteria Analytical sensitivity vCJD brain spikes in human plasma (WHO reference number NHBY0/0003) 24 replicates of each of three dilutions of the material WHO number NHBY0/0003 (1×10 4 , 1×10 5 , 1×10 6 ) 23 of the 24 replicates detected at 1×10 4 vCJD spleen spikes in human plasma (10 % spleen homogenate — NIBSC reference number NHSY0/0009) 24 replicates of each of three dilutions of the material NIBSC number NHSY0/0009 (1×10, 1×10 2 , 1×10 3 ) 23 of the 24 replicates detected at 1×10 Diagnostic sensitivity A) Specimen from appropriate animal models As many specimen as reasonably possible and available, and at least 10 specimens 90 % B) Specimen from humans with known clinical vCJD As many specimen as reasonably possible and available, and at least 10 specimens 90 % Only in case where 10 specimens are not available: the number of specimens tested shall be comprised between 6 and 9 all available specimens shall be tested no more than one false negative result Analytical specificity Potentially cross-reacting blood-specimens 100 Diagnostic specificity Normal human plasma samples from area of low BSE exposure 5 000 At least 99,5 % --- Source: https://www.legislation.gov.uk/eudn/2002/364/annex/division/3 (the official version) Page: https://openlegislation.uk/eu-decision-2002-364/annex/division-3 Contains public sector information licensed under the Open Government Licence v3.0. Source: legislation.gov.uk, The National Archives.