EN   EN  
 
 
 EUROPEAN  
COMMISSION   
Brussels, XXX  
[…](2018) XXX  draft  
ANNEX  
 
ANNEX  
to the  
Commission implementing Decision  
amending Decision 2002/364/EC as regards requirements for HIV and HCV antigen and 
antibody combined tests and as regards requirements for nucleic acid amplification 
techniques with respect to reference materials and qualitative HIV assays  EN 1  EN ANNEX  
 
The Annex is amended as follows:  
 
(1) Sub-section 3.1.1. is replaced by the following:  
‘3.1.1 Devices which detect virus infections shall meet the requirements for sensitivity and 
specificity set out in Table 1 and Table 5  according to virus type and entities detected (antigen  
and/or antibody) . See also principle 3.1.11 for screening assays. ’ 
 
(2) Section 3.2. is replaced by the following:  
‘3.2. Additio nal requirements for HIV and HCV antigen and antibody combined tests.  
 
3.2.1. HIV antigen  and antibody combined tests intended for the detection of HIV -1 p24 
antigen  and HIV -1/2 antibody shall meet  the requirements  for sensitivity and specificity  set 
out in Table 1  and Table 5 .  
 
3.2.2. Hepatitis C virus (HC V) antigen  and antibody combined tests intended for the detection  
of HCV antigen  and HCV antibody shall meet  the requirements for sensitivity and specificity 
set out in Table 1  and Table 5 . 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). HCV  antig en and antibody combined tests shall demonstrate enhanced 
sensitivity in early HCV  infection when compared to HCV antibody only tests. ’  
 
(3) Sub-section 3.3.2. is replaced by the following:  
‘3.3.2. The analytical sensitivity or detection limit for NAT  assays shall be expressed by the 
95 % positive cut -off value. This is the analyte concentration where 95  % of test runs give 
positive results following serial dilutions of an international reference material, where 
available, such as a World Health Organisation ( WHO ) International Standard or reference 
material calibrated against a WHO International Standard.’  
 
(4) The following sub-sections 3.3.2a and 3.3.2b are inserted:  
‘3.3.2a. Qualitative HIV NAT assays 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. 
3.3.2b. Qualitative HIV NAT assays, other than virus typing assays, shall be designed to 
compensate  for the potential failure of a HIV -1 NAT target region, e.g. by using two 
independent target regions.’  
 
(5) Table 1 is replaced by the following:   
EN 2  EN   
‘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 Notified 
Body or manufacturer)  To be defined 
when available  20 panels  
10 further panels (at Notified Body or 
manufacturer)  
 20 panels  
10 further panels (at 
Notified 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  
EN 3  EN  
(6) Table 5 is replaced by the following:  
 
                                                 
1 The total number of seroconversion panels for combined Ag/Ab assays (from tables 1 and 5) need not be greater than 30.  ‘Table 5:  
HIV 1 antigen , HIV Ag/Ab,  HCV antigen , HCV Ag/Ab  
 
 HIV -1 antigen and HIV Ag/Ab assays  HCV antigen  and HCV Ag/Ab assay s 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 panels1 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 
principle s in § 3.1.5