Commission Implementing Regulation (EU) 2022/1107 of 4 July 2022 laying down common specifications for certain class D in vitro diagnostic medical devices in accordance with Regulation (EU) 2017/746 of the European Parliament and of the Council (Text with EEA relevance)

5.7.2022	EN	Official Journal of the European Union	L 178/3

THE EUROPEAN COMMISSION,

Having regard to the Treaty on the Functioning of the European Union,

Having regard to Regulation (EU) 2017/746 of the European Parliament and of the Council of 5 April 2017 on in vitro diagnostic medical devices and repealing Directive 98/79/EC and Commission Decision 2010/227/EU ¹, and in particular Article 9(1) thereof,

Whereas:

• (1) For certain class D in vitro diagnostic medical devices falling within the scope of Regulation (EU) 2017/746, harmonised standards do not exist as regards certain requirements of Annex I to that Regulation, and there is a need to address public health concerns as the risk associated with the use of those devices is significant for public health and patient safety. It is therefore appropriate to adopt common specifications for those devices in respect of those requirements.

• (2) Regulation (EU) 2017/746 replaces Directive 98/79/EC of the European Parliament and of the Council ². The common technical specifications set out in Commission Decision 2002/364/EC ³ for certain devices covered by Directive 98/79/EC remain relevant. Those common technical specifications have therefore been taken into account and where necessary updated to reflect the state of the art.

• (3) To allow manufacturers, other economic operators, notified bodies and other actors to adapt to this Regulation, and to ensure its proper application, it is appropriate to defer its application. However, in the interest of public health and patient safety, manufacturers should be allowed to comply with the common specifications laid down in this Regulation on a voluntary basis before its date of application.

• (4) To ensure a continuous high level of safety and performance of devices, as a transitional measure it should be provided that devices that are in conformity with Decision 2002/364/EC are to be presumed to be in conformity with the requirements for certain performance characteristics set out in Annex I to Regulation (EU) 2017/746 until the date of application of this Regulation.

• (5) The Medical Device Coordination Group has been consulted.

• (6) The measures provided for in this Regulation are in accordance with the opinion of the Committee on Medical Devices,

HAS ADOPTED THIS REGULATION:

Article 1

Common specifications

This Regulation lays down common specifications for certain class D in vitro diagnostic medical devices in respect of the requirements regarding the performance characteristics set out in Section 9.1, points (a) and (b), Section 9.3 and Section 9.4, point (a), of Annex I to Regulation (EU) 2017/746.

Annex I lays down common specifications for devices covered by Annexes II to XIII, as specified in that Annex.

Annex II lays down common specifications for devices intended for detection of blood group antigens in the ABO, Rh, Kell, Duffy and Kidd blood group systems.

Annex III lays down common specifications for devices intended for detection or quantification of markers of human immunodeficiency virus (HIV) infection.

Annex IV lays down common specifications for devices intended for detection or quantification of markers of human T-cell lymphotropic virus (HTLV) infection.

Annex V lays down common specifications for devices intended for detection or quantification of markers of hepatitis C virus (HCV) infection.

Annex VI lays down common specifications for devices intended for detection or quantification of markers of hepatitis B virus (HBV) infection.

Annex VII lays down common specifications for devices intended for detection or quantification of markers of hepatitis D virus (HDV) infection.

Annex VIII lays down common specifications for devices intended for detection of markers of variant Creutzfeldt-Jakob disease (vCJD).

Annex IX lays down common specifications for devices intended for detection or quantification of markers of cytomegalovirus (CMV) infection.

Annex X lays down common specifications for devices intended for detection or quantification of markers of Epstein-Barr virus infection (EBV).

Annex XI lays down common specifications for devices intended for detection of markers of Treponema pallidum infection.

Annex XII lays down common specifications for devices intended for detection or quantification of markers of Trypanosoma cruzi infection.

Annex XIII lays down common specifications for devices intended for detection or quantification of markers of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection.

Article 2

Definitions

For the purposes of this Regulation, the following definitions apply:

• (1) ‘true positive’ means a specimen known to be positive for the target marker and correctly classified by the device;

• (2) ‘false negative’ means a specimen known to be positive for the target marker and misclassified by the device;

• (3) ‘false positive’ means a specimen known to be negative for the target marker and misclassified by the device;

• (4) ‘the limit of detection’ (‘LOD’) means the smallest amount of the target marker that can be detected;

• (5) ‘nucleic acid amplification techniques’ (‘NAT’) means methods of detection and/or quantification of nucleic acids by either amplification of a target sequence, by amplification of a signal or by hybridisation;

• (6) ‘NAT system’ means the combination of devices used for extraction, amplification and detection of nucleic acids;

• (7) ‘rapid test’ means a qualitative or semi-quantitative in vitro diagnostic medical device, used singly or in a small series, which involves non-automated procedures (except the reading of results) and has been designed to give a fast result;

• (8) ‘robustness’ means the capacity of an analytical procedure to remain unaffected by small but deliberate variations in method parameters and provides an indication of its reliability during normal usage;

• (9) ‘cross-reactivity’ means the ability of non-target analytes or markers to cause false positive results in an assay because of similarity, e.g. the ability of non-specific antibodies binding to a test antigen of an antibody assay, or the ability of non-target nucleic acids to be reactive in a NAT assay;

• (10) ‘interference’ means the ability of unrelated substances to affect the results in an assay;

• (11) ‘whole system failure rate’ means the frequency of failures when the entire process is performed as prescribed by the manufacturer;

• (12) ‘first-line assay’ means a device used to detect a marker or analyte, and the use of which may be followed by the use of a confirmatory assay; devices intended solely to be used to monitor a previously determined marker or analyte are not considered first-line assays;

• (13) ‘confirmatory assay’ means a device used for the confirmation of a reactive result from a first line assay;

• (14) ‘supplemental assay’ means a device that is used to provide further information for the interpretation of the test result of another assay;

• (15) ‘virus typing device’ means a device used for typing with already known positive samples, not used for primary diagnosis of infection or for screening;

• (16) ‘95 % positive cut-off value’ means the analyte concentration where 95 % of test runs give positive results following serial dilutions of an international reference material, where available, e.g. a World Health Organisation (WHO) International Standard or reference material calibrated against the WHO International Standard.

Article 3

Transitional provisions

During that period manufacturers of devices that are not in conformity with the common technical specifications set out in Decision 2002/364/EC shall duly justify that they have adopted solutions that ensure a level of safety and performance that is at least equivalent thereto.

Article 4

Entry into force and date of application

This Regulation shall enter into force on the twentieth day following that of its publication in the Official Journal of the European Union.

It shall apply from 25 July 2024.

However, Article 3 shall apply from 25 July 2022.

This Regulation shall be binding in its entirety and directly applicable in all Member States.

Done at Brussels, 4 July 2022.

For the Commission

The President

Ursula VON DER LEYEN

ANNEX I

GENERAL COMMON SPECIFICATIONS

Part I – Requirements for performance characteristics of devices covered by Annexes II to XIII

Performance characteristics	Requirement
All performance characteristics set out in Section 9.1, points (a) and (b), Section 9.3 and Section 9.4, point (a), of Annex I to Regulation (EU) 2017/746	1. The determination of performance characteristics shall be carried out in direct comparison with a state-of-the-art device. The device used for comparison shall be one bearing CE marking, if on the market at the time of the performance evaluation. 2. Devices used for determination of status of specimens used in determination of performance characteristics shall be state-of-the-art devices bearing CE marking. 3. If discrepant results are identified as part of determination of performance characteristics, these results shall be resolved as far as possible, by one or more of the following: — by evaluation of the discrepant specimen in further devices, — by use of an alternative method or marker, — by a review of the clinical status and diagnosis of the patient, — by the testing of follow-up specimens. 4. The determination of performance characteristics shall be performed on a population equivalent to the European population.
Whole system failure rate	5. As part of the required risk analysis the whole system failure rate leading to false negative results shall be determined in repeat assays on low-positive specimens.
Analytical sensitivity and analytical specificity, interference	6. For devices intended for use with plasma the manufacturer shall verify the performance of the device using all anticoagulants which the manufacturer indicates for use with the device, for at least 50 plasma specimens (for devices intended for detection and/or quantification of infectious agents, 25 positive and 25 negative).
Analytical and diagnostic specificity, interference and cross-reactivity	7. The manufacturer shall select the potential interfering substances to be evaluated taking account of the composition of the reagents and configuration of the device.
Batch-to-batch consistency	8. For devices intended to detect antigens and antibodies, the manufacturer’s batch testing criteria shall ensure that every batch consistently identifies the relevant antigens, epitopes, and antibodies and is suitable for the claimed specimen types. 9. The manufacturer’s batch release testing for first-line assays shall include at least 100 specimens negative for the relevant analyte4.

Part II – Requirements for performance characteristics of devices referred to in Annexes III to XIII

Performance characteristic	Requirement
Analytical and diagnostic sensitivity	10. Devices intended by the manufacturer for testing body fluids other than serum or plasma, e.g. urine, saliva, etc., shall meet the same requirements as serum or plasma devices. The manufacturer shall test specimens from the same individuals in both the devices to be approved and in a respective serum or plasma device.5 11. Devices for self-testing shall meet the same requirements as respective devices for professional use. 12. Positive specimens used in the performance evaluation shall be selected to reflect different stages of the respective disease(s), different antibody patterns, different genotypes, different subtypes, mutants, etc. 13. Seroconversion panels shall start with a negative bleed(s) and shall have narrow bleeding intervals as far as possible. Where this is not possible, manufacturers shall provide a justification in the performance evaluation report. 14. For devices intended by the manufacturer to be used with serum and plasma the performance evaluation must demonstrate serum to plasma equivalency. This shall be demonstrated for at least 25 positive donations. 15. For devices detecting or quantifying antigens or nucleic acids, the target antigen(s) or target nucleic acid region(s) respectively shall be specified in the instructions for use. 16. For devices detecting or quantifying antibodies against an infectious agent, the target antigen(s) of those antibodies shall be specified in the instructions for use.
Analytical and diagnostic specificity	17. Devices intended by the manufacturer for testing body fluids other than serum or plasma, e.g. urine, saliva, etc., shall meet the same requirements as serum or plasma devices. The performance evaluation shall test specimens from the same individuals in both the devices to be approved and in a respective serum or plasma device.5 18. Devices for self-testing shall meet the same requirements as respective devices for professional use. 19. Negative specimens used in a performance evaluation shall be defined so as to reflect the target population for which the device is intended, such as blood donors, hospitalised patients, pregnant women, etc. 20. Specificity shall be based on repeatedly reactive false positive results in specimens negative for the target marker. 21. For devices intended by the manufacturer to be used with serum and plasma the performance evaluation must demonstrate serum to plasma equivalency. This shall be demonstrated for at least 25 negative donations.
Analytical and diagnostic specificity, interference and cross-reactivity	22. The manufacturer shall include specimens such as, where applicable: — specimens representing related infections, — specimens from multigravida, i.e. women who have had more than one pregnancy, or rheumatoid factor (RF) positive patients, — specimens containing human antibodies to components of the expression system, for example anti-E. coli, or anti-yeast.
Performances obtained by lay persons	23. Relevant parts of the performance evaluation shall be carried out (or repeated) by appropriate lay persons to validate the operation of the device and the instructions for use. The lay persons selected for the performance evaluation shall be representative of the intended users groups.

ANNEX II

COMMON SPECIFICATIONS FOR DEVICES INTENDED FOR DETECTION OF BLOOD GROUP ANTIGENS IN THE ABO, RH, KELL, DUFFY AND KIDD BLOOD GROUP SYSTEMS

Scope

This Annex applies to devices intended for detection of blood group antigens in the ABO, Rh, Kell, Duffy and Kidd blood group systems.

Table 1 applies to performance evaluation of devices detecting blood group antigens in the ABO, Rh, Kell, Duffy and Kidd blood group systems.

Table 2 applies to manufacturer’s batch-to-batch consistency testing of reagents and reagent products to determine blood group antigens in the ABO, Rh, Kell, Duffy and Kidd blood group systems (test reagents, control materials).

Table 1. Performance evaluation of devices detecting blood group antigens in the ABO, Rh, Kell, Duffy and Kidd blood group systems

Reagent specificity	Number of tests per method claimed by the manufacturer	Total number of specimens to be tested for a launch device	Total number of specimens to be tested for a new formulation, or use of well-characterised reagents	General qualification criteria	Specific qualification criteria	Acceptance criteria
Anti-ABO1 (Anti-A), Anti-ABO2 (Anti-B), Anti-ABO3 (Anti-A,B)	≥500	≥3 000	≥1 000	Clinical specimens: 10 % of the test population Neonatal specimens: > 2 % of the test population	ABO specimens shall include > 40 % A and B antigen positive specimens which may include specimens from group A, group B and group AB	All of the reagents shall show comparable performance to state-of-the-art CE marked devices with regard to claimed reactivity of the device. For CE marked devices where the application or use has been changed or extended, further testing shall be carried out in accordance with the requirements outlined in column 2 above (“Number of tests per method claimed by the manufacturer”).
Anti-RH1 (Anti-D)	≥500	≥3 000	≥1 000	Clinical specimens: 10 % of the test population Neonatal specimens: > 2 % of the test population	Performance evaluation of Anti-D reagents shall include tests against a range of weak RH1 (D) and partial RH1 (D) specimens, depending on the intended use of the product. Weak and/or partial D cells shall account for > 2 % of RH1 (D) positive specimens.	All of the reagents shall show comparable performance to state-of-the-art CE marked devices with regard to claimed reactivity of the device. For CE marked devices where the application or use has been changed or extended, further testing shall be carried out in accordance with the requirements outlined in column 2 above (“Number of tests per method claimed by the manufacturer”).
Anti-RH2 (Anti-C), Anti-RH4 (Anti-c), Anti- RH3 (Anti-E)	≥100	≥1 000	≥200	Clinical specimens: 10 % of the test population Neonatal specimens: > 2 % of the test population		All of the reagents shall show comparable performance to state-of-the-art CE marked devices with regard to claimed reactivity of the device. For CE marked devices where the application or use has been changed or extended, further testing shall be carried out in accordance with the requirements outlined in column 2 above (“Number of tests per method claimed by the manufacturer”).
Anti-RH5 (Anti-e)	≥100	≥500	≥200	Clinical specimens: 10 % of the test population Neonatal specimens: > 2 % of the test population		All of the reagents shall show comparable performance to state-of-the-art CE marked devices with regard to claimed reactivity of the device. For CE marked devices where the application or use has been changed or extended, further testing shall be carried out in accordance with the requirements outlined in column 2 above (“Number of tests per method claimed by the manufacturer”).
Anti-KEL1 (Anti-K)	≥100	≥500	≥200	Clinical specimens: 10 % of the test population Neonatal specimens: > 2 % of the test population		All of the reagents shall show comparable performance to state-of-the-art CE marked devices with regard to claimed reactivity of the device. For CE marked devices where the application or use has been changed or extended, further testing shall be carried out in accordance with the requirements outlined in column 2 above (“Number of tests per method claimed by the manufacturer”).
Anti-JK1 (Jka), Anti-JK2 (Jkb)	≥100	≥500	≥200	Clinical specimens: 10 % of the test population Neonatal specimens: > 2 % of the test population		All of the reagents shall show comparable performance to state-of-the-art CE marked devices with regard to claimed reactivity of the device. For CE marked devices where the application or use has been changed or extended, further testing shall be carried out in accordance with the requirements outlined in column 2 above (“Number of tests per method claimed by the manufacturer”).
Anti-FY1 (Fya), Anti-FY2 (Fyb)	≥100	≥500	≥200	Clinical specimens: 10 % of the test population Neonatal specimens: > 2 % of the test population		All of the reagents shall show comparable performance to state-of-the-art CE marked devices with regard to claimed reactivity of the device. For CE marked devices where the application or use has been changed or extended, further testing shall be carried out in accordance with the requirements outlined in column 2 above (“Number of tests per method claimed by the manufacturer”).
Note: Positive specimens used in the performance evaluation shall be selected to reflect variant and weak antigen expression.	Note: Positive specimens used in the performance evaluation shall be selected to reflect variant and weak antigen expression.	Note: Positive specimens used in the performance evaluation shall be selected to reflect variant and weak antigen expression.	Note: Positive specimens used in the performance evaluation shall be selected to reflect variant and weak antigen expression.	Note: Positive specimens used in the performance evaluation shall be selected to reflect variant and weak antigen expression.	Note: Positive specimens used in the performance evaluation shall be selected to reflect variant and weak antigen expression.	Note: Positive specimens used in the performance evaluation shall be selected to reflect variant and weak antigen expression.

Table 2. Manufacturer’s batch-to-batch consistency testing of reagents and reagent products to determine blood group antigens in the ABO, Rh, Kell, Duffy and Kidd blood group systems

1. Test reagents

Blood group reagents	Minimum number of control cells to be tested as part of specificity testing	Minimum number of control cells to be tested as part of specificity testing	Minimum number of control cells to be tested as part of specificity testing	Minimum number of control cells to be tested as part of specificity testing	Minimum number of control cells to be tested as part of specificity testing	Minimum number of control cells to be tested as part of specificity testing	Minimum number of control cells to be tested as part of specificity testing	Minimum number of control cells to be tested as part of specificity testing	Acceptance criteria
	Positive reactions	Positive reactions	Positive reactions	Positive reactions		Negative reactions	Negative reactions	Negative reactions	Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
	A1	A2B	Ax			B	O		Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
Anti-ABO1(Anti-A)	2	2	26			2	2		Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
	B	A1B				A1	O		Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
Anti-ABO2(Anti-B)	2	2				2	2		Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
	A1	A2	Ax	B		O			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
Anti-ABO3(Anti-A,B)	2	2	26	2		4			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
	R1r	R2r	WeakD			r’r	r”r	rr	Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
Anti-RH1 (Anti-D)	2	2	26			1	1	1	Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
	R1R2	R1r	r’r			R2R2	r”r	rr	Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
Anti-RH2 (Anti-C)	2	1	1			1	1	1	Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
	R1R2	R1r	r’r			R1R1			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
Anti-RH4 (Anti-c)	1	2	1			3			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
	R1R2	R2r	r”r			R1R1	r’r	rr	Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
Anti-RH3 (Anti-E)	2	1	1			1	1	1	Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
	R1R2	R2r	r”r			R2R2			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
Anti-RH5 (Anti-e)	2	1	1			3			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
	Kk					kk			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
Anti-KEL1 (Anti-K)	4					3			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
	Jk(a+b+)					Jk(a–b+)			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
Anti-JK1 (Anti-Jka)	4					3			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
	Jk(a+b+)					Jk(a+b–)			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
Anti-JK2 (Anti-Jkb)	4					3			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
	Fy(a+b+)					Fy(a–b+)			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
Anti-FY1 (Anti-Fya)	4					3			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
	Fy(a+b+)					Fy(a+b–)			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
Anti-FY2 (Anti-Fyb)	4					3			Each batch of reagent shall exhibit unequivocal positive or negative results by all techniques claimed by the manufacturer in accordance with the results obtained from the performance evaluation data.
Note: Polyclonal reagents shall be tested against a wider panel of cells to confirm specificity and exclude presence of unwanted contaminating antibodies.	Note: Polyclonal reagents shall be tested against a wider panel of cells to confirm specificity and exclude presence of unwanted contaminating antibodies.	Note: Polyclonal reagents shall be tested against a wider panel of cells to confirm specificity and exclude presence of unwanted contaminating antibodies.	Note: Polyclonal reagents shall be tested against a wider panel of cells to confirm specificity and exclude presence of unwanted contaminating antibodies.	Note: Polyclonal reagents shall be tested against a wider panel of cells to confirm specificity and exclude presence of unwanted contaminating antibodies.	Note: Polyclonal reagents shall be tested against a wider panel of cells to confirm specificity and exclude presence of unwanted contaminating antibodies.	Note: Polyclonal reagents shall be tested against a wider panel of cells to confirm specificity and exclude presence of unwanted contaminating antibodies.	Note: Polyclonal reagents shall be tested against a wider panel of cells to confirm specificity and exclude presence of unwanted contaminating antibodies.	Note: Polyclonal reagents shall be tested against a wider panel of cells to confirm specificity and exclude presence of unwanted contaminating antibodies.	Note: Polyclonal reagents shall be tested against a wider panel of cells to confirm specificity and exclude presence of unwanted contaminating antibodies.

2. Control materials (red cells)

The phenotype of red cells used in the control of blood typing reagents listed above shall be confirmed using an established device(s).

ANNEX III

COMMON SPECIFICATIONS FOR DEVICES INTENDED FOR DETECTION OR QUANTIFICATION OF MARKERS OF HUMAN IMMUNODEFICIENCY VIRUS (HIV) INFECTION

Scope

	1.	This Annex applies to devices intended for detection or quantification of markers of human immunodeficiency virus (HIV) infection. Table 1 applies to first-line assays for HIV-1/2 antibody (anti-HIV-1/2) and first-line combined antigen/antibody assays for HIV-1/2 (HIV-1/2 Ag/Ab) which are not rapid tests. Table 2 applies to first-line assays for anti-HIV-1/2 and HIV-1/2 Ag/Ab which are rapid tests. Table 3 applies to confirmatory assays for anti-HIV-1/2. Table 4 applies to antigen tests for HIV-1 and HIV Ag/Ab assays. Table 5 applies to qualitative and quantitative NAT devices for HIV ribonucleic acid (RNA). Table 6 applies to HIV-1/2 self-tests.

Definitions

	2.	For the purposes of this Annex, the following definitions apply: (1) ‘seroconversion HIV specimen’ means: — p24 antigen and/or HIV RNA positive, and — recognised by the antibody first-line assays, and — positive or indeterminate in confirmatory assays. (2) ‘early seroconversion HIV specimen’ means: — p24 antigen and/or HIV RNA positive, and — not recognised by the antibody first-line assays, and — indeterminate or negative in confirmatory assays. Table 1. First-line assays: anti-HIV-1/2, HIV-1/2 Ag/Ab (requirements for antibody detection) Performance characteristic Specimen Specimen numbers, features, use Acceptance criteria Diagnostic sensitivity Positive specimens ≥400 HIV-1 ≥100 HIV-2 including 40 non-B-subtypes including 25 positive ‘same day’ fresh serum specimens (≤ 1 day after specimen taking) all available HIV/1 subtypes shall be represented by at least 3 specimens per subtype all true positive specimens shall be identified as positive Seroconversion panels ≥30 panels at least 40 early seroconversion HIV specimens shall be tested diagnostic sensitivity during seroconversion shall represent the state of the art all seroconversion HIV specimens shall be identified as positive Diagnostic specificity Unselected blood donors (including first-time donors)7 ≥5 000 ≥99,5% Hospitalised patients ≥200 Potential limitations for specificity, if any, shall be identified Cross-reactivity Potentially cross-reacting specimens ≥100 in total (such as RF+, from related virus infections, from pregnant women, subjects recently vaccinated against any infectious agent) Table 2. Rapid tests: anti-HIV-1/2, HIV-1/2 Ag/Ab (requirements for antibody detection) Performance characteristic Specimen Specimen numbers, features, use Acceptance criteria 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 specimens per subtype all true positive specimens shall be identified as positive Seroconversion panels ≥30 panels at least 40 early seroconversion HIV specimens shall be tested diagnostic sensitivity during seroconversion shall represent the state of the art all seroconversion HIV specimens shall be identified as positive Diagnostic specificity Unselected blood donors (including first-time donors) ≥1 000 ≥ 99 % Hospitalised patients ≥200 Potential limitations for specificity, if any, shall be identified Cross-reactivity Potentially cross-reacting specimens ≥200 specimens from pregnant women ≥100 other potentially cross-reacting specimens in total (e.g. RF+, from related infections) Table 3. Confirmatory assays: anti-HIV-1/2 Performance characteristic Specimen Specimen numbers, features, use Acceptance criteria Diagnostic sensitivity Positive specimens ≥200 HIV-1 ≥100 HIV-2 Including different stages of infection and reflecting different antibody patterns Identification as “confirmed positive” or “indeterminate”, not as “negative” Seroconversion panels ≥15 seroconversion panels/low titre panels ≥40 early seroconversion HIV specimens Diagnostic sensitivity during seroconversion shall represent the state of the art All seroconversion HIV specimens shall be identified as positive Diagnostic specificity Blood donors ≥200 No false positive results / no neutralisation Hospitalised patients ≥200 Cross-reactivity Potentially cross-reacting specimens ≥50 in total (including specimens from pregnant women, specimens with indeterminate results in other confirmatory assays) Table 4. Antigen tests: HIV-1, HIV Ag/Ab (requirements for antigen detection) Performance characteristic Specimen Specimen numbers, features, use Acceptance criteria Diagnostic sensitivity Positive specimens ≥50 HIV-1 antigen positive ≥50 cell culture supernatants including different HIV-1 subtypes and HIV-2 all true positive specimens shall be identified as positive (after neutralisation if applicable) Seroconversion panels ≥20 seroconversion panels/low titre panels ≥40 early seroconversion HIV specimens diagnostic sensitivity during seroconversion shall represent the state of the art all seroconversion HIV specimens shall be identified as positive Analytical sensitivity First International Reference Reagent HIV-1 p24 Antigen, NIBSC code: 90/636 ≤ 2 IU/ml Diagnostic specificity Blood donors ≥200 ≥ 99,5 % after neutralisation or, if no neutralisation test available, after resolution of the specimen status Hospitalised patients ≥200 Potential limitations for specificity, if any, shall be identified Cross-reactivity Potentially cross-reacting specimens ≥50

Table 5. Qualitative and quantitative NAT devices for HIV RNA

	1.	For target sequence amplification devices, a functionality control for each specimen (internal control) shall reflect the state of the art. This control shall as far as possible be used throughout the whole process, i.e. extraction, amplification/hybridisation, detection.
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	2.	Genotype and/or subtype detection shall be demonstrated by appropriate primer or probe design validation and shall also be validated by testing characterised genotyped specimens.
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	3.	Potential cross-reactivity of non-target nucleic acid sequences shall be analysed by appropriate primer or probe design validation and shall also be validated by testing selected specimens.
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	4.	Results of quantitative NAT devices shall be traceable to international standards or calibrated reference materials, if available, and be expressed in international units utilised in the specific field of application.
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	5.	Qualitative HIV NAT devices intended to be used to detect the presence of HIV in blood, blood components, cells, tissues or organs, or in any of their derivatives, in order to assess their suitability for transfusion, transplantation or cell administration shall be designed to detect both HIV-1 and HIV-2.
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	6.	Qualitative HIV NAT devices, other than virus typing devices, shall be designed to compensate for the potential failure of a HIV-1 NAT target region by using two independent target regions. Performance characteristic Specimen Specimen numbers, features, use Acceptance criteria Analytical sensitivity WHO International Standard HIV-1 RNA; WHO International Standard HIV-2 RNA; or calibrated reference materials NAT sensitivity and NAT LOD shall be validated by dilution series of reference materials, testing of replicates (minimum 24) at different analyte concentrations, including those with transition from positive to negative results with the respective NAT device. LOD shall be expressed as 95% positive cut-off value (IU/ml) after statistical analysis (e.g. Probit).8 quantitative NAT: definition of lower, upper quantification limit, precision, accuracy, ‘linear’ measuring range, ‘dynamic range’. Reproducibility at different concentration levels According to the state of the art HIV geno-/subtype sensitivity all relevant genotypes/subtypes, preferably from international reference materials potential substitutes for rare HIV subtypes (to be quantified by appropriate methods): cell culture supernatants; in vitro transcripts; plasmids Qualitative NAT: at least 10 specimens/genotype or subtype Quantitative NAT: dilution series for demonstration of quantification efficiencies According to the state of the art Diagnostic sensitivity Positive specimens reflecting the routine conditions of users (e.g. no pre-selection of specimens) Quantitative NAT: ≥100 Comparative results with another NAT system shall be generated in parallel According to the state of the art Seroconversion panels Qualitative NAT: ≥10 panels Comparative results with another NAT system shall be generated in parallel According to the state of the art Diagnostic specificity Blood donor specimens Qualitative NAT: ≥500 Quantitative NAT: ≥100 According to the state of the art Cross-reactivity Potentially cross-reacting specimens ≥10 human retrovirus positive specimens (e.g. HTLV) According to the state of the art Carry-over High HIV RNA positive; HIV RNA negative At least five runs with alternating high-positive and negative specimens shall be performed during robustness studies. The virus titres of the high positive specimens shall be representative of high virus titres occurring naturally. According to the state of the art Detection in relation to antibody status HIV-RNA positives: anti-HIV negative, anti-HIV positive Pre-seroconversion (anti-HIV negative) and post-seroconversion (anti-HIV positive) specimens According to the state of the art Whole system failure rate HIV RNA low-positive ≥100 HIV RNA low-positive specimens shall be tested. These specimens shall contain a virus concentration equivalent to three times the 95 % positive cut-off virus concentration. ≥99% positive Table 6. Additional requirements for HIV-1/2 self-tests Performance characteristic Specimens9 Number of lay persons Result interpretation10 Interpretation of results11 by lay persons reflecting the following range of reactivity levels: — non-reactive — reactive — weak reactive12 — invalid ≥ 100 Diagnostic sensitivity lay persons that are known positive ≥ 200 Diagnostic specificity lay persons that do not know their status ≥ 400 Lay persons that are at high risk of acquiring the infection ≥ 200

ANNEX IV

COMMON SPECIFICATIONS FOR DEVICES INTENDED FOR DETECTION OR QUANTIFICATION OF MARKERS OF HUMAN T-CELL LYMPHOTROPIC VIRUS (HTLV) INFECTION

Scope

This Annex applies to devices intended for detection or quantification of markers of human T-cell lymphotropic virus (HTLV) infection.

Table 1 applies to first-line assays for antibodies against HTLV I or II (anti-HTLV I/II) which are not rapid tests.

Table 2 applies to first-line assays for anti-HTLV I/II which are rapid tests.

Table 3 applies to confirmatory assays for anti-HTLV I/II.

Table 4 applies to NAT devices for HTLV I/II.

Table 1. First-line assays: anti-HTLV I/II

Performance characteristic	Specimen	Specimen numbers, features, use	Acceptance criteria
Diagnostic sensitivity	Positive specimens	≥300 HTLV-I ≥100 HTLV-II including 25 positive ‘same day’ fresh serum specimens (≤ 1 day after specimen taking)	all true positive specimens shall be identified as positive
Diagnostic sensitivity	Seroconversion panels	To be defined when available	diagnostic sensitivity during seroconversion shall represent the state of the art, if applicable
Diagnostic specificity	Unselected blood donors (including first-time donors)13	≥5 000	≥ 99,5%
Diagnostic specificity	Hospitalised patients	≥200	Potential limitations for specificity, if any, shall be identified
Cross-reactivity	Potentially cross-reacting specimens	≥100 in total (e.g. RF+, from related virus infections, from pregnant women)	Potential limitations for specificity, if any, shall be identified

Table 2. Rapid tests: anti-HTLV I/II

Performance characteristic	Specimen	Specimen numbers, features, use	Acceptance criteria
Diagnostic sensitivity	Positive specimens	≥300 HTLV-I ≥100 HTLV-II	all true positive specimens shall be identified as positive
Diagnostic sensitivity	Seroconversion panels	To be defined when available	diagnostic sensitivity during seroconversion shall represent the state of the art, if applicable
Diagnostic specificity	Unselected blood donors (including first-time donors)	≥1 000	≥ 99%
Diagnostic specificity	Hospitalised patients	≥200	Potential limitations for specificity, if any, shall be identified
Cross-reactivity	Potentially cross-reacting specimens	≥200 specimens from pregnant women ≥100 other potentially cross-reacting specimens in total (e.g. RF+, from related infections)	Potential limitations for specificity, if any, shall be identified

Table 3. Confirmatory assays: anti-HTLV I/II

Performance characteristic	Specimen	Specimen numbers, features, use	Acceptance criteria
Diagnostic sensitivity	Positive specimens	≥200 HTLV I ≥100 HTLV II	Identification as “confirmed positive” or “indeterminate”, not as “negative”
Diagnostic sensitivity	Seroconversion panels	To be defined when available	diagnostic sensitivity during seroconversion shall represent the state of the art, if applicable
Diagnostic specificity	Blood donors	≥200	No false positive results
Diagnostic specificity	Hospitalised patients	≥200	No false positive results
Cross-reactivity	Potentially cross-reacting specimens	≥50 in total (including specimens from pregnant women, specimens with indeterminate results in other confirmatory assays)	No false positive results

Table 4. NAT devices for HTLV I/II

	1.	For target sequence amplification devices, a functionality control for each specimen (internal control) shall reflect the state of the art. This control shall as far as possible be used throughout the whole process, i.e. extraction, amplification/hybridisation, detection.
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	2.	Genotype and/or subtype detection shall be demonstrated by appropriate primer or probe design validation and shall also be validated by testing characterised genotyped specimens.
---	---	---
	3.	Potential cross-reactivity of non-target nucleic acid sequences shall be analysed by appropriate primer or probe design validation and shall also be validated by testing selected specimens.
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	4.	Results of quantitative NAT devices shall be traceable to international standards or calibrated reference materials, if available, and be expressed in international units utilised in the specific field of application. Performance characteristic Specimen Specimen numbers, features, use Acceptance criteria Analytical sensitivity International reference preparations NAT sensitivity and NAT LOD shall be validated by dilution series of reference materials, testing of replicates (minimum 24) at different analyte concentrations, including those with transition from positive to negative results with the respective NAT device. LOD shall be expressed as 95% positive cut-off value (IU/ml) after statistical analysis (e.g. Probit).14 quantitative NAT: definition of lower, upper quantification limit, precision, accuracy, ‘linear’ measuring range, ‘dynamic range’. Reproducibility at different concentration levels According to the state of the art HTLV I and HTLV II genotype sensitivity all relevant genotypes, preferably from international reference materials potential substitutes for rare HTLV genotypes (to be quantified by appropriate methods): cell culture supernatants; in vitro transcripts; plasmids Qualitative NAT: at least 10 specimens/genotype or subtype Quantitative NAT: dilution series for demonstration of quantification efficiencies According to the state of the art Diagnostic specificity Blood donor specimens Qualitative NAT: ≥500 Quantitative NAT: ≥100 According to the state of the art Cross-reactivity Potentially cross-reacting specimens ≥10 human retrovirus positive specimens (e.g. HIV-1, HIV-2) According to the state of the art Carry-over High HTLV RNA positive; HTLV RNA negative At least five runs with alternating high-positive and negative specimens shall be performed during robustness studies. The virus titres of the high positive specimens shall be representative of high virus titres occurring naturally. According to the state of the art Detection in relation to antibody status HTLV-RNA positives: anti-HTLV negative, anti-HTLV positive Pre-seroconversion (anti-HTLV negative) and post-seroconversion (anti-HTLV positive) specimens According to the state of the art Whole system failure rate HTLV RNA low-positive ≥100 HTLV RNA low-positive specimens shall be tested. These specimens shall contain a virus concentration equivalent to three times the 95 % positive cut-off virus concentration. ≥99% positive

ANNEX V

COMMON SPECIFICATIONS FOR DEVICES INTENDED FOR DETECTION OR QUANTIFICATION OF MARKERS OF HEPATITIS C VIRUS (HCV) INFECTION

Scope

This Annex applies to devices intended for detection or quantification of markers of hepatitis C virus (HCV) infection.

Table 1 applies to first-line assays for anti-HCV antibodies (anti-HCV) and combined antigen/antibody tests for HCV (HCV Ag/Ab) which are not rapid tests.

Table 2 applies to first-line assays for anti-HCV and HCV Ag/Ab which are rapid tests.

Table 3 applies to confirmatory and supplemental assays for anti-HCV.

Table 4 applies to HCV antigen tests and HCV Ag/Ab.

Table 5 applies to qualitative and quantitative NAT devices for HCV RNA.

Table 6 applies to HCV self-tests.

Table 1. First-line assays: anti-HCV, HCV Ag/Ab (requirements for antibody detection)

Performance characteristic	Specimen	Specimen numbers, features, use	Acceptance criteria
Diagnostic sensitivity	Positive specimens	≥400 Including specimens from different stages of infection and reflecting different antibody patterns HCV genotype 1-4: > 20 specimens per genotype (including non-a subtypes of genotype 4); HCV genotypes 5 and 6: > 5 specimens each; including 25 positive ‘same day’ fresh serum specimens (≤ 1 day after specimen taking)	all true positive specimens shall be identified as positive
	Seroconversion panels	≥30 panels HCV seroconversion panels for the evaluation of HCV antigen and antibody combined tests (HCV Ag/Ab) shall start with one or more negative bleeds and comprise panel members from early HCV infection (HCV core antigen and/or HCV RNA positive but anti-HCV negative).	diagnostic sensitivity during seroconversion shall represent the state of the art HCV Ag/Ab tests shall demonstrate enhanced sensitivity in early HCV infection when compared to HCV antibody only tests.
Diagnostic specificity	Unselected blood donors (including first-time donors)15	≥5 000	≥99,5%
Diagnostic specificity	Hospitalised patients	≥200	Potential limitations for specificity, if any, shall be identified
Cross-reactivity	Potentially cross-reacting specimens	≥100 in total (e.g. RF+, from related virus infections, from pregnant women)	Potential limitations for specificity, if any, shall be identified

Table 2. Rapid tests: anti-HCV, HCV Ag/Ab (requirements for antibody detection)

Performance characteristic	Specimen	Specimen numbers, features, use	Acceptance criteria
Diagnostic sensitivity	Positive specimens	≥400 including specimens from different stages of infection and reflecting different antibody patterns. HCV genotype 1-4: > 20 specimens per genotype (including non-a subtypes of genotype 4); HCV genotypes 5 and 6: > 5 specimens each;	all true positive specimens shall be identified as positive
Diagnostic sensitivity	Seroconversion panels	≥30 panels HCV seroconversion panels for the evaluation of HCV antigen and antibody combined tests (HCV Ag/Ab) shall start with one or more negative bleeds and comprise panel members from early HCV infection (HCV core antigen and/or HCV RNA positive but anti-HCV negative).	diagnostic sensitivity during seroconversion shall represent the state of the art HCV Ag/Ab tests shall demonstrate enhanced sensitivity in early HCV infection when compared to HCV antibody only tests.
Diagnostic specificity	Unselected blood donors (including first-time donors)1	≥1 000	≥ 99 %
Diagnostic specificity	Hospitalised patients	≥200	Potential limitations for specificity, if any, shall be identified
Cross-reactivity	Potentially cross-reacting specimens	≥200 specimens from pregnant women ≥100 other potentially cross-reacting specimens in total (e.g. RF+, from related infections)	Potential limitations for specificity, if any, shall be identified

Table 3. Confirmatory and supplemental assays: anti-HCV

Performance characteristic	Specimen	Specimen numbers, features, use	Acceptance criteria
Diagnostic sensitivity	Positive specimens	≥300 Including specimens from different stages of infection and reflecting different antibody patterns. HCV genotypes 1 – 4: > 20 specimens (including non-a subtypes of genotype 4; HCV genotype 5: > 5 specimens; HCV genotype 6: as far as available	identification as “confirmed positive” or “indeterminate”, not as “negative”
Diagnostic sensitivity	Seroconversion panels	≥15 seroconversion panels/low titre panels	diagnostic sensitivity during seroconversion shall represent the state of the art
Diagnostic specificity	Blood donors	≥200	No false positive results/ no neutralisation
Diagnostic specificity	Hospitalised patients	≥200	No false positive results/ no neutralisation
Cross-reactivity	Potentially cross-reacting specimens	≥50 in total (including specimens from pregnant women, specimens with indeterminate results in other confirmatory assays)	No false positive results/ no neutralisation

Table 4. Antigen tests: HCV antigen, HCV Ag/Ab (requirements for antigen detection)

Performance characteristic	Specimen	Specimen numbers, features, use	Acceptance criteria
Diagnostic sensitivity	Positive specimens	≥25 HCV core antigen and/or HCV RNA positive but anti-HCV negative specimens, comprising HCV genotypes 1-6 (if a genotype is not available, a justification shall be made)	all true positive specimens shall be identified as positive
Diagnostic sensitivity	Seroconversion panels	≥20 seroconversion panels/low titre panels HCV seroconversion panels for the evaluation of HCV antigen and antibody combined tests shall start with one or more negative bleeds and comprise panel members from early HCV infection (HCV core antigen and/or HCV RNA positive but anti-HCV negative).	diagnostic sensitivity during seroconversion shall represent the state of the art HCV antigen and antibody combined tests shall demonstrate enhanced sensitivity in early HCV infection when compared to HCV antibody only tests.
Analytical sensitivity	WHO International Standard HCV core (PEI 129096/12)	Dilution series
Diagnostic specificity	Blood donors	≥200	≥ 99,5 % after neutralisation or, if no neutralisation test available, after resolution of the specimen status
Diagnostic specificity	Hospitalised patients	≥200	Potential limitations for specificity, if any, shall be identified
Cross-reactivity	Potentially cross-reacting specimens	≥50	Potential limitations for specificity, if any, shall be identified

Table 5. Qualitative and quantitative NAT devices for HCV RNA

	1.	For target sequence amplification devices, a functionality control for each specimen (internal control) shall reflect the state of the art. This control shall as far as possible be used throughout the whole process, i.e. extraction, amplification/hybridisation, detection.
---	---	---
	2.	Genotype and/or subtype detection shall be demonstrated by appropriate primer or probe design validation and shall also be validated by testing characterised genotyped specimens.
---	---	---
	3.	Potential cross-reactivity of non-target nucleic acid sequences shall be analysed by appropriate primer or probe design validation and shall also be validated by testing selected specimens.
---	---	---
	4.	Results of quantitative NAT devices shall be traceable to international standards or calibrated reference materials, if available, and be expressed in international units utilised in the specific field of application. Performance characteristic Specimen Specimen numbers, features, use Acceptance criteria Analytical sensitivity WHO International Standard HCV RNA(or calibrated reference materials) NAT sensitivity and NAT LOD shall be validated by dilution series of reference materials, testing of replicates (minimum 24) at different analyte concentrations, including those with transition from positive to negative results with the respective NAT device. LOD shall be expressed as 95% positive cut-off value (IU/ml) after statistical analysis (e.g. Probit).16 quantitative NAT: definition of lower, upper quantification limit, precision, accuracy, ‘linear’ measuring range, ‘dynamic range’. Reproducibility at different concentration levels According to the state of the art HCV genotype sensitivity all relevant genotypes/subtypes, preferably from international reference materials potential substitutes for rare HCV genotypes (to be quantified by appropriate methods): in vitro transcripts; plasmids Qualitative NAT: ≥10 specimens/genotype or subtype Quantitative NAT: dilution series for demonstration of quantification efficiencies According to the state of the art Diagnostic sensitivity Positive specimens reflecting the routine conditions of users (e.g. no pre-selection of specimens) Quantitative NAT: ≥100 Comparative results with another NAT system shall be generated in parallel According to the state of the art Seroconversion panels Qualitative NAT: ≥10 panels Comparative results with another NAT system shall be generated in parallel According to the state of the art Diagnostic specificity Blood donor specimens Qualitative NAT: ≥500 Quantitative NAT: ≥100 According to the state of the art Cross-reactivity Potentially cross-reacting specimens >10 human flavivirus (e.g. HGV, YFV) positive specimens According to the state of the art Carry-over High HCV RNA positive; HCV RNA negative At least five runs with alternating high-positive and negative specimens shall be performed during robustness studies. The virus titres of the high positive specimens shall be representative of high virus titres occurring naturally. According to the state of the art Detection in relation to antibody status HCV RNA positives: anti-HCV negative, anti-HCV positive Pre-seroconversion (anti-HCV negative) and post-seroconversion (anti-HCV positive) specimens According to the state of the art Whole system failure rate HCV RNA low-positive ≥100 HCV RNA low-positive specimens shall be tested. These specimens shall contain a virus concentration equivalent to three times the 95 % positive cut-off virus concentration. ≥99% positive

Table 6. Additional requirements for HCV self-tests

Performance characteristic	Specimens17	Number of lay persons
Result interpretation18	Interpretation of results19 by lay persons reflecting the following range of reactivity levels: — non-reactive — reactive — weak reactive20 — invalid	≥ 100
Diagnostic sensitivity	lay persons that are known positive	≥ 200
Diagnostic specificity	lay persons that do not know their status	≥ 400
Diagnostic specificity	lay persons that are at high risk of acquiring the infection	≥ 200

ANNEX VI

COMMON SPECIFICATIONS FOR DEVICES INTENDED FOR DETECTION OR QUANTIFICATION OF MARKERS OF HEPATITIS B VIRUS (HBV) INFECTION

Scope

This Annex applies to devices intended for detection or quantification of markers of hepatitis B virus (HBV) infection.

Table 1 applies to first-line assays for hepatitis B surface antigen (HBsAg), and for antibodies against hepatitis B core antigen (anti-HBc) which are not rapid tests.

Table 2 applies to first-line assays for HBsAg and anti-HBc which are rapid tests.

Table 3 applies to confirmatory assays for HBsAg.

Table 4 applies to assays for the hepatitis B virus markers: hepatitis B surface antibodies (anti-HBs), IgM antibody against the hepatitis B core antigen (anti-HBc IgM), antibodies against the hepatitis Be antigen (anti-HBe) and hepatitis Be antigen (HBeAg).

Table 5 applies to qualitative and quantitative NAT devices for HBV deoxyribonucleic acid (DNA).

Table 6 applies to HBV self-tests.

Table 1. First-line assays: HBsAg, anti-HBc

Performance characteristic	Specimen	Specimen numbers, features, use	Acceptance criteria
Diagnostic sensitivity	Positive specimens	≥400 anti-HBc: including evaluation of different HBV markers HBsAg: including different HBV genotypes / subtypes / mutants anti-HBc or HBsAg: including 25 positive ‘same day’ fresh serum (≤ 1 day after specimen taking)	Overall performance shall be at least equivalent to the comparator device
Diagnostic sensitivity	Seroconversion panels	HBsAg assays: ≥30 panels anti-HBc assays: to be defined when available	diagnostic sensitivity during seroconversion shall represent the state of the art (for anti-HBc this shall be the case if applicable)
Analytical sensitivity	WHO Third International Standard HBsAg (subtypes ayw1/adw2, HBV genotype B4, NIBSC code: 12/226)		For HBsAg assays: <0,130 IU/ml
Diagnostic specificity	Unselected blood donors (including first-time donors)21	≥5 000	≥99,5%
Diagnostic specificity	Hospitalised patients	≥200	Potential limitations for specificity, if any, shall be identified
Cross-reactivity	Potentially cross-reacting specimens	≥100 in total (e.g. RF+, from related virus infections, from pregnant women,)	Potential limitations for specificity, if any, shall be identified

Table 2. Rapid tests: HBsAg, anti-HBc

Performance characteristic	Specimen	Specimen numbers, features, use	Acceptance criteria
Diagnostic sensitivity	Positive specimens	≥400 including evaluation of different HBV markers including different HBV genotypes / subtypes / mutants	Overall performance shall be at least equivalent to that of the comparator device
Diagnostic sensitivity	Seroconversion panels	HBsAg assays: ≥30 panels anti-HBc assays: to be defined when available	Diagnostic sensitivity during seroconversion shall represent the state of the art (for anti-HBc this shall be the case if applicable)
Diagnostic specificity	Unselected blood donors (including first-time donors)	≥1 000	HBsAg assays: ≥ 99 % anti-HBc assays: ≥ 99 %
Diagnostic specificity	Hospitalised patients	≥200	Potential limitations for specificity, if any, shall be identified
Cross-reactivity	Potentially cross-reacting specimens	≥200 specimens from pregnant women ≥100 other potentially cross-reacting specimens in total (e.g. RF+, from related infections)	Potential limitations for specificity, if any, shall be identified

Table 3. Confirmatory assays: HBsAg

Performance characteristic	Specimen	Specimen numbers, features, use	Acceptance criteria
Diagnostic sensitivity	Positive specimens	≥300 Including specimens from different stages of infection Including 20 ‘high positive’ specimens (>26 IU/ml); 20 specimens in the cut-off range	Correct identification as positive (or indeterminate), not negative
Diagnostic sensitivity	Seroconversion panels	≥15 seroconversion panels/low titre panels	Diagnostic sensitivity during seroconversion shall represent the state of the art
Analytical sensitivity	WHO Third International Standard for HBsAg, subtypes ayw1/adw2, HBV genotype B4, NIBSC code: 12/226
Diagnostic specificity	Negative specimens	≥10 false positives as available from the performance evaluation of the first-line assay	No false positive results/ no neutralisation
Cross-reactivity	Potentially cross-reacting specimens	≥50	No false positive results/ no neutralisation

Table 4. Assays for the HBV markers: anti-HBs, anti-HBc IgM, anti-HBe, HBeAg