Sensitivity of individual donor nucleic acid testing (NAT) for the detection of hepatitis B infection by studying diluted NAT yield samples.

BACKGROUND Screening blood donors for the presence of hepatitis B virus surface antigen (HBsAg) has been the backbone of blood safety. However, occult hepatitis B infection (OBI) in donors can be missed when only HBsAg screening is used. Nucleic acid testing (NAT) is capable of detecting OBI among donors. The aim of our study was to analyse the sensitivity of NAT for detecting OBI. MATERIAL AND METHODS The kits used during the study for serology testing were BioRad Monolisa™ HBsAg Ultra (HBsAg screening), Abbott Architect for anti-HBcAg (total) and anti-HBsAg testing, and Vitros® by Ortho Clinical Diagnostics for anti-HBcAg (IgM). Procleix Ultrio was used for individual donor-NAT (ID-NAT) and Abbott m2000 for estimation of HBV DNA. Out of 28,134 HBsAg non-reactive donors, 25 were ID-NAT-reactive. Of these 25 NAT yield samples, 18 were studied further at different dilutions from 1:2 to 1:16. The doubling dilutions were made with HBV non-reactive AB plasma. Undiluted samples were used for all serological tests and for HBV DNA estimation. RESULTS Of the 18 samples studied, nine were NAT-reactive at a dilution of <1:4 and five out of these showed presence of antibody to core antigen (IgG+IgM). Antibody to surface antigen was present in only two of the nine NAT-reactive samples, one with antibody to core antigen and the other without. Six had a viral load in the range from <10 to 38 IU/mL whereas the viral load in the remaining three samples was not determined. Among the other nine samples which were NAT-reactive at dilutions≥1:4, antibody to core antigen (IgG+IgM) was present in seven. DISCUSSION Our study showed that ID-NAT testing along with HBsAg screening could detect most potentially HBV infectious donors (including those with OBI). NAT screening for HBV on diluted samples could compromise blood safety because samples with a low viral load will escape detection.