S.L. 427.16 Art. 7secrecy vis-a-vis the competent carried out) effect. The following For the background in - if -

The staff of the inspection body are bound to observe professional with regard to all information gained in the course of their duties (except administrative authorities of the State in which their activities are under these regulations or any provision of national law putting it into

SCHEDULE X Based on Annex X of the Directive

CE MARKING OF CONFORMITY CE conformity marking shall consist of the initials "CE" taking the form:

avoidance of doubt, it is hereby decdared that the grid providing the the above graduated drawing is not part of the CE marking. the marking is reduced or enlarged the proportions given in the above graduated drawing must be respected, the various components of the CE marking must have substantially the same vertical dimension, which may not be less than 5mm. This minimum dimension may be waived for small-scale devices.

SCHEDULE XI

Annex I of Commission Decision 2009/108/EC of 3 February 2009 on common technical specifications (CTS) for in vitro diagnostic medical devices

SCOPE common technical specifications set out in this Schedule shall apply for of Schedule II, List A. DEFINITIONS AND TERMS (Diagnostic) sensitivity

The probability that the device gives a positive result in the target marker.

True positive A specimen known to be positive for the target correctly classified by the device.

False negative A specimen known to be positive for the target misclassified by the device.

(Diagnostic) specificity The probability that the device gives a in the absence of the target marker.

False positive A specimen known to be negative for the target marker and misclassified the device. True negative A specimen known to be negative for the target marker classified by the device. Analytical sensitivity

Analytical sensitivity may be expressed as the limit of detection, i.e. amount of the target marker that can be precisely detected.

Analytical specificity Analytical specificity means the ability of the method to determine target marker. Nucleic acid amplification techniques (NAT) The term ‘NAT’ is used for tests for the detection and/or quantification nucleic acids by either amplification of a target sequence, by amplification signal or by hybridisation. Rapid test ‘Rapid test’ means qualitative or semi quantitative in vitro medical devices, used singly or in a small series, which involve procedures and have been designed to give a fast result. Robustness The robustness of an analytical procedure means the capacity of procedure to remain unaffected by small but deliberate variations parameters and provides an indication of its reliability during normal Whole system failure rate The whole system failure rate means the frequency of failures entire process is performed as prescribed by the manufacturer. Confirmation assay

Confirmation assay means an assay used for the confirmation of a from a screening assay.

Virus typing assay Virus typing assay means an assay used for typing with already known samples, not used for primary diagnosis of infection or for screening. Sero-conversion HIV samples Sero-conversion HIV samples mean:

p24 antigen and/or HIV RNA positive and; recognised by all of the antibody screening tests and; positive or indeterminate confirmatory assays. sero-conversion HIV samples sero-conversion HIV samples mean:

p24 antigen and/or HIV RNA positive and; not recognised by all of the antibody screening tests and; indeterminate or negative confirmatory assays. COMMON TECHNICAL SPECIFICATIONS (CTS) FOR PRODUCTS TO IN SCHEDULE II, LIST A. for performance evaluation of reagents and reagent products for the confirmation and quantification in human specimens of markers of HIV 1 and 2), HTLV I and II, and hepatitis B, C, D:

Principles: Devices which detect virus infections placed on the market for use as screening or diagnostic tests, shall meet the requirements for sensitivity and out in Table 1. See also principle 3.1.11 for screening assays.

Devices intended by the manufacturer for testing body fluids other than plasma, for example urine, saliva, etc shall meet the same CTS for sensitivity and specificity as serum or plasma tests. The evaluation shall test samples from the same individuals in both the tests and in a respective serum or plasma assay.

Devices intended by the manufacturer for self-test, i.e. home use, shall CTS requirements for sensitivity and specificity as respective devices professional use. Relevant parts of the performance evaluation shall be (or repeated) by appropriate lay users to validate the operation of the instructions for use.

performance evaluations shall be carried out in direct comparison with state of the art device. The device used for comparison shall be CE marking, if on the market at the time of the performance discrepant test results are identified as part of an evaluation, these be resolved as far as possible, for example: evaluation of the discrepant sample in further test systems; use of an alternative method or marker; a review of the clinical status and diagnosis of the patient; and the testing of follow-up-samples. Performance evaluations shall be performed on a population the European population. Positive specimens used in the performance evaluation shall be reflect different stages of the respective disease(s), different patterns, different genotypes, different subtypes, mutants, etc. Sensitivity with true positives and sero-conversion samples shall be evaluated as follows:

3.1.8.1 Diagnostic test sensitivity during sero-conversion has to state of the art. Whether further testing of the same or additional sero-conversion panels is conducted by the notified body or by the manufacturer shall confirm the initial performance evaluation data (see Seroconversion panels should start with a negative bleed(s) and should bleeding intervals. 3.1.8.2 For blood screening devices (with the exception of HBsAg HBc tests), all true positive samples shall be identified as positive to be CE marked (Table 1). For HBsAg and anti-HBc tests the new have an overall performance at least equivalent to that of the established (see 3.1.4). 3.1.8.3 Regarding HIV tests:

- all sero-conversion HIV samples shall be identified as positive; - at least 40 early sero-conversion HIV samples shall be should conform to the state of the art.

3.1.9 Performance evaluation of screening assays shall include 25 available in the case of rare infections) "same day" fresh serum samples (≤ 1 day after sampling). 3.1.10 Negative specimens used in a performance evaluation shall so as to reflect the target population for which the test is intended, for donors, hospitalised patients, pregnant women, etc. 3.1.11 For performance evaluations for screening assays (Table 1) populations shall be investigated from at least two blood donation consist of consecutive blood donations, which have not been selected first time donors. 3.1.12 Devices shall have a specificity of at least 99.5% on blood unless otherwise indicated in the accompanying tables. Specificity calculated using the frequency of repeatedly reactive (i.e. false results in blood donors negative for the target marker.

3.1.13 Devices shall be evaluated to establish the effect interfering substances, as part of the performance evaluation. interfering substances to be evaluated will depend to some extent composition of the reagent and configuration of the assay. Potential substances shall be identified as part of the risk analysis required essential requirements for each new device but may include, for example: - specimens representing "related" infections; - specimens from multipara, i.e. women who have had one pregnancy, or rheumatoid factor positive patients; - for recombinant antigens, human antibodies to components expression system, for example anti E. coli, or anti yeast. 3.1.14 For devices intended by the manufacturer to be used with plasma the performance evaluation must demonstrate serum equivalency. This shall be demonstrated for at least 50 donations (25 negative). 3.1.15 For devices intended for use with plasma the evaluation shall verify the performance of the device using all which the manufacturer indicates for use with the device. This shall be donations (25 positive and 25 negative).

part of the required risk analysis the whole system failure rate false-negative results shall be determined in repeat assays on low-specimens. new in vitro diagnostic medical device belonging to Schedule II, List specifically covered by the common technical specification, the technical specification for a related device should be taken into account. devices may be identified on different grounds, for example by the similar intended use or by similar risks. Additional Requirements for HIV antibody/antigen combined tests antibody/antigen combined tests intended for anti-HIV and p24 detection which include claims for single p24 antigen detection shall follow Table 5, including criteria for analytical sensitivity for p24 antigen. antibody/antigen combined tests intended for anti-HIV and p24 which do not include claims for single p24 detection shall follow Table 5, excluding criteria for analytical sensitivity for p24. Additional Requirements for Nucleic Acid Amplification Techniques performance evaluation criteria for NAT assays can be found in Table 2. target sequence amplification assays, a functionality control for sample (internal control) shall reflect the state of the art. This control as possible be used throughout the whole process, i.e. extraction, amplification/hybridisation, detection.

analytical sensitivity or detection limit for NAT assays shall be the 95% positive cut-off value. This is the analyte concentration where runs give positive results following serial dilutions of an international material for example a WHO standard or calibrated reference material. Genotype detection shall be demonstrated by appropriate primer or probe validation and shall also be validated by testing characterised genotyped

Results of quantitative NAT assays shall be traceable to international calibrated reference materials, if available, and be expressed in units utilised in the specific field of application. assays may be used to detect virus in antibody negative samples, conversion samples. Viruses within immune-complexes may differently in comparison to free viruses, for example during a step. It is therefore important that during robustness studies, antibody-negative (pre sero-conversion) samples are included. investigation of potential carry-over, at least five runs with alternating and negative specimens shall be performed during robustness high positive samples shall comprise of samples with naturally virus titres.

whole system failure rate leading to false-negative results shall be by testing low-positive specimens. Low positive specimens shall concentration equivalent to 3 times the 95% positive cut-off virus for the manufacturer’s release testing of reagents and eagent products for the detection, confirmation and quantification in human specimens HIV infection (HIV 1 and 2), HTLV I and II, and hepatitis B, C, D (Immunological assays only). 3.4.1 The manufacturer’s release testing criteria shall ensure that consistently identifies the relevant antigens, epitopes, and antibodies. 3.4.2. The manufacturer’s batch release testing for screening include at least 100 specimens negative for the relevant analyte. 3.5 CTS for performance evaluation of reagents and reagent determining the following blood group antigens: ABO blood group (A), ABO2 (B), ABO3 (A,B); Rh blood group system RH1 (D), RH2 RH4 (c), RH5 (e); Kell blood group system KEL1 (K). Criteria for performance evaluation of reagents and reagent determining the blood groups antigens: ABO blood group system ABO2 (B), ABO3 (A,B); Rh blood group B 9 system RH1 (D), RH2 RH4 (c), RH5 (e); Kell blood group system KEL1 (K) can be found in

3.5.1 All performance evaluations shall be carried out in direct comparison an established state of the art device. The device used for comparison one bearing CE marking, if on the market at the time of the evaluation. 3.5.2 If discrepant test results are identified as part of an evaluation, results shall be resolved as far as possible, for example: - by evaluation of the discrepant sample in further test - by use of an alternative method. 3.5.3 Performance evaluations shall be performed on equivalent to the European population. 3.5.4 Positive specimens used in the performance evaluation shall to reflect variant and weak antigen expression. 3.5.5 Devices shall be evaluated to establish the effect interfering substances, as part of the performance evaluation. interfering substances to be evaluated will depend to some extent composition of the reagent and configuration of the assay. Potential substances shall be identified as part of the risk analysis required essential requirements for each new device. 3.5.6 For devices intended for use with plasma the performance evaluation verify the performance of the device using all anticoagulants manufacturer indicates for use with the device. This shall be demonstrated least 50 donations. 3.6. CTS for the manufacturers release testing of reagents and reagent for determining the blood group antigens: ABO blood group system ABO2 (B), ABO3 (A,B); Rh blood group system RH1 (D), RH2 (C), (c), RH5 (e); Kell blood group system KEL1 (K).

3.6.1 The manufacturer’s release testing criteria shall ensure that consistently identifies the relevant antigens, epitopes, and antibodies.

3.6.2 Requirements for manufacturers batch release testing are outlined 10.

Table 1: "Screening" assays: anti-HIV 1 and 2, anti-HTLV I and II, HBsAg, anti-HBc anti-HIV-1/2 anti-HTLV- anti-HCV HBsAg anti-HBc I/II Positive 400HIV-1100 300 HTLV-I 400 (positive 400 Including 400 specimens HIV-2 100 HTLV-II samples) subtype- Including including 40 consideration evaluation Including non-B- of other samples from subtypes, all HBV-available HIV/ different stages markers of infection and 1 subtypes reflecting should be represented by different antibody at least 3 patterns. samples per subtype Genotype 1-4: > 20 samples - per genotype (includingnon-a sub-types of genotype 4);

5: > 5 samples;

6: if available Sero- 20 panels To be 20 panels 20 panels To be conversion 10 further defined 10 further panels 10 further defined panels panels (at when (at Notified panels (at when Notified Body available Body or NotifiedBody available or manufacturer) or manufacturer) manufacturer) Standards 0.130 IU/ml (Second International Standard for HBsAg, subtypeadw2, genotype A, NIBSC code: 00/588) Unselected 5000 5000 5000 5000 5000 donors (including1 time donors) Hospitalize 200 200 200 200 200 patients Potentially 100 100 100 100 100 cross-reacting blood-specimens (RF+, related viruses, pregnant women,

Table 2: NAT assays for HIV1, HCV, HBV, HTLV I/II (qualitative quantitative; not molecular typing)

HIV1 HCV HBV HTLV

NAT qualitative quantitative qualitative quantitative quantitative qualitative

AsforHIV As for HIV quantitative quantitative Sensitivity According Detection According According According Detection to EP limit: as to EP to EP to EP limit validation for validation validation validation Detection guideline qualitative guideline guideline guideline 1 1 1 of (1): tests; ( ): ( ):several ( ): several analytical several Quantific- several dilution dilution sensitivity dilution ationlimit: dilution series into series into (IU/ml; seriesinto dilutions series into borderline borderline definedon borderline (half- borderline concentr- concentr-WHO concentra- log10 or concentra- ation; ation; standards tion; less) of tion; statistical statistical or statistical calibrated statistical analysis analysis calibrated analysis reference analysis (e.g. (e.g. Probit reference (e.g. prepara- (e.g. Probit analysis) materials) Probit tions, Probit analysis) onthebasis analysis) definition analysis) on the of at least on the of lower, on the basis of at 24 basis of at upper basis of at least 24 replicates; least 24 quantifi- least 24 replicates; calculation replicates; cation replicates; calculation of 95% cut-calculatio limit, calculatio of 95% off value n of 95% precision, n of 95% cut-off cut-off accuracy, cut-off value value "linear" value measuring range, "dynamic range". Reproduci -bility at different concentra-tion levels to be shown Genotype/ At least 10 Dilution Atleast10 As far as As far as subtype samplesper series of samples calibrated calibrated detection/ subtype (as all per genotype genotype quantific- far as relevant genotype reference reference ation available) genotypes (as far as materials materials efficiency /subtypes, available) are are preferabl available available y of reference materials, as far as available HCV HBV HTLV I/II Accep-tance qualitative quantitative qualitative quantitative quantitative qualitative quantitative criteria AsforHIV As for HIV AsforHIV quantitative quantitative quantitative culture Transcrip supernatant ts or plasmids quantified by appropria te According meethods may be validation used

According According According far as to EP to EP to EP

validation validation validation guideline guideline guideline

(1)asfaras (1)asfaras (1) as far calibrated calibrated as subtype subtype calibrated reference reference subtype materials materials reference transcripts are are materials bean available; available; are in vitro in vitro available; transcripts transcripts in vitro couldbean couldbean transcripts option option could be an option blood 100 500 blood 500 blood 500 blood blood donors donors donors donors

As for By assays By assays By assays qualitative design and/ design and/ design tests or testing ortestingof and/or of at least at least 10 testing of 10 human other DNA- at least 10 flavivirus virus human (e.g. HGV, positive retrivirus YFV) samples (e.g.HIV-) and/ positive positive testing samples samples least human retrovirus

As for qualitativ e tests 5 At least 5 At least 5 At least 5 using runs using runs using runsusing alternatin alternating alternating alternatin high high g high positive positive positive to (known to (known to (knownto occur occur occur naturally) naturally) naturally) naturally) and and and negative negative negative samples samples samples Internal Internal Internal control control control preferably preferably preferably preferably to go to go to go through the throughthe through whole whole NAT wholeNAT the whole procedure procedure NAT procedure procedure HIV1 HCV HBV HTLV

NAT qualitative quantitative qualitative quantitative quantitative qualitative

AsforHIV As for HIV quantitative quantitative Whole At least At least At least At least system 100 100 100 100 failurerate samples samples samples samples leading to virus- virus- virus- virus-false-neg spikedwith spiked spiked spiked results 3 x the with 3 x with 3 x with 3 x 95% pos the 95% the 95 % the 95 % cut-off poscut-off poscut-off poscut-off concentra- concentra- concentra- concentr-tion tion tion ation

(1) European Pharmacopoeia guideline

Note: Acceptance criteria for ‘whole system failure rate leading to false-neg assays positive.

For quantitative NATs a study shall be performed on at least 100 reflecting the routine conditions of users (e.g. no pre-selection of specimens). results with another NAT test system shall be generated in parallel.

For qualitative NATs a study on diagnostic sensitivity shall be performed sero-conversion panels.

Comparative results with another NAT test system shall be generated in parallel. Table 3: Rapid tests: anti-HIV 1 and 2, anti-HCV, HBsAg, HTLV I and II

anti-HIV 1/2 anti-HCV HBsAg anti-HBc anti-HTLV II Diagnostic Positive Same criteria Same criteria Same criteria Same criteria Same criteria sensitivity specimens as for as for as for as for as for screening screening screening screening screening assays assays assays assays assays Sero- Same criteria Same criteria Same criteria Same criteria Same criteria conversion as for as for as for as for as for panels screening screening screening screening screening assays assays assays assays assays

Negative 1000 blood 1000 blood 1000 blood 1000 blood 1000 blood specimens donations 200 donations donations donations donations clinical 200 clinical 200 clinical 200 clinical 200 clinical specimens specimens200 specimens200 specimens specimens 200 samples samples from samples from 200 samples from pregnant pregnant pregnant frompregnant women women women women 100potentially 100potentially 100 100 100 interfering interfering potentially potentially potentially samples samples interfering interfering interfering samples samples samples Confirmatory/supplementary assays for anti-HIV 1 and 2, anti-HTLV anti-HCV, HBsAg anti-HIV anti-HTLV HCV HBsAg Acceptance Confirmatory Confirmatory Supplementary Confirmatory criteria Assay Assay Assay Assay Positive 200HIV-1and 200 HTLV-I 300 HCV 300 HBsAg Correct specimens 100 HIV-2 and 100 (positive identification HTLV-II samples) as positive (or indeterminate), not negative Including Including Including samples from samples from samples from different different different stages stages of stages of of infection 20 infection and infection and “high pos” reflecting reflecting samples (> 26 different different IU/ml); 20 antibody antibody samples in the patterns patterns. cut-off range Genotypes 1 -4: > 20 samples (including non-a sub-types of genotype 4); 5: > 5 samples; 6: if available Sero-con- 15 sero- 15 15 version conversion seroconversion seroconversion panels panels/ low panels/ low panels/ low titre panels titre panels titre panels Standards Second International Standard for HBsAg, subtype adw2, genotype A, NIBSC code: 00/588 anti-HIV anti-HTLV HCV HBsAg Confirmatory Confirmatory Supplementary Confirmatory Assay Assay Assay Assay Diagnostic Negative 200 blood 200 blood 200 blood 10 false specificity specimens donations donations donations positives as availablefrom the performance evaluation of the screening assay. (1) 200 clinical 200 clinical 200 clinical samples samples samples including including including pregnant pregnant pregnant women women women

50 50 50potentially 50 potentially potentially potentially interfering interfering interfering interfering samples, samples samples, samples, including including including samples with samples with samples with indeterminate indetermin- indeterminat results in ate results in e results in other other other confirmatory confirmatory confirmatory assays assays assays Table 5: HIV 1 Antigen

HIV-1 Antigen Assay Acceptance Diagnostic Positive specimens 50 HIV-1 Ag-positive Correct identification sensitivity 50 cell culture supernatants neutralisation) including different HIV-1 subtypes and HIV-2 Sero-conversion panels 20 sero-conversion panels/low titre panels Analytical Standards HIV-1 p24 Antigen, 1st ≤ 2 IU/ml sensitivity International Reference Reagent, NIBSC code: 90/636 Diagnostic 200 blood donations 200 >99.5% after specificity clinical samples 50 potentially interfering samples

Table 6: Serotyping and Genotyping Assay: HCV

HCV Serotyping and Acceptance Genotyping Assay Diagnostic Positive specimens 200 (positive samples) > 95% agreement sensitivity Including samples from serotyping different stages of infection and > 95% agreement reflecting different antibody genotyping patterns. Genotypes 1 - 4: > 20 samples (including non-a sub-types of genotype 4); 5: > 5 samples; 6: if available HCV Serotyping and Acceptance criteria Genotyping Assay Negative specimens 100

Markers: anti-HBs, anti HBc IgM, anti-HBe, HBeAg anti-HBs anti-HBc IgM anti-HBe HBeAg Acceptance criteria Positive 100 vaccinees 200 Including 200 Including 200 Including > 98% specimens 100 naturally samples from samples from samples from infected different stages different stages different persons of infection of infection stages of (acute/chronic (acute/chronic infection etc.) etc.) (acute/chronic etc.) The acceptance criteria should only be applied on samples from acute infection stage Sero-con- 10 follow- When available version ups or anti-panels HBs sero-conversions Standards WHO 1st HBe - Anti-HBs: International Referenzan- <10 mIU/ml Reference tigen 82; PEI Preparation Germany 1977; NIBSC, United Kingdom 500 200 blood 200 blood 200 blood >98% negative Including donations donations donations specimens clinical samples 200 clinical 200 clinical 200 clinical samples samples samples 50 potentially 50 potentially 50 potentially interfering interfering interfering samples samples samples HDV markers: anti-HDV, anti-HDV IgM, Delta Antigen anti-HDV anti-HDV IgM Delta Antigen Acceptance criteria Positive 100 50 >98% specimens Specifying HBV- Specifying HBV-markers markers Negative 200 200 200 >98% specimens Including clinical Including clinical Including clinical samples samples samples

50 potentially 50 potentially 50 potentially interfering samples interfering samples interfering samples Table 9: Blood group antigens in the ABO, Rh and Kell blood group systems

1 2 Specificity Number of tests per Total number of samples Total recommended to be tested for a launch to be method product formulation, well-characterized

Anti-ABO1(anti-A),anti- 500 3000 1000 Anti-ABO2 (anti-B), Anti-ABO3 (anti-A,B) Anti-RH1 (anti-D) 500 3000 1000 Anti-RH2 (anti-C ), anti- 100 1000 200 RH4 (anti-c), anti-RH3 (anti-E) Anti-RH5 (anti-e) 100 500 200 Anti-KEL1 (anti-K) 100 500 200 Acceptance criteria: All of the above reagents shall show comparable test established reagents with acceptable performance with regard to claimed of the device. For established reagents, where the application or changed or extended, further testing should be carried out in accordance requirements outlined in column 1 (above). Performance evaluation of anti-D-reagents shall include tests against weak RH1 (D) and partial RH1 (D) samples, depending on the intended product. Qualifications: Clinical samples: 10% of the test population Neonatal specimens: > 2% of the test population ABO samples: > 40% A, B positives ‘weak D’: > 2% of RH1 (D) positives

Table 10: Batch release criteria for reagents and reagent determine blood group antigens in the ABO, Rh and Kell blood group

Specificity Testing Requirements on each reagent