Evaluation of 17 CE-marked HBsAg assays with respect to clinical sensitivity, analytical sensitivity, and hepatitis B virus mutant detection

Abstract

Seventeen HBsAg assays, in use in the European market (CE-marked), were assessed for their diagnostic sensitivity using 38 commercially available seroconversion panels, and for their analytical sensitivity with the HBsAg ad and ay standards of the Paul-Ehrlich-Institut (PEI). In addition, the ability to detect HBsAg mutants was investigated by means of 21 recombinant HBsAg mutant samples and 5 natural mutants. Analysis of seroconversion data revealed that there were marked differences in the sensitivity among the CE-marked HBsAg assays. Differences in the window period between the most and the least sensitive assays were up to 2 weeks. Analytical sensitivities of the investigated assays ranged from 0.009 to 0.05 PEI-U/ml for HBsAg ad standard (relating to approximately 0.018 to 0.100 IU/ml of the 2nd WHO HBsAg standard) and 0.012 to 0.11 PEI-U/ml for the ay standard. Clinical and analytical sensitivities were basically correlated. The capacity to detect mutant HBsAg forms was influenced by the assay format and the properties of the monoclonal antibodies used for coating of the solid phase or in the conjugate. While some assays detected all mutants others exhibited weaknesses especially in recognising HBsAg mutations affecting loop 2 of the HBsAg a-determinant. The results obtained with the recombinant mutants were largely confirmed by the investigation of clinical samples. The study gives a broad overview of the current state of the art of about 70% of the HBsAg assays currently available in Europe. The overall sensitivity has not been improved further since 1995 when the most sensitive assay was introduced into the market. In addition, detection of HBsAg mutants seems problematic with several assays. It is concluded that there is potential to improve clinical sensitivity and mutant recognition of HBsAg assays.

Detection of Highly Prevalent Hepatitis B Virus Coinfection among HIV-Seropositive Persons in Ghana

“…Previous studies indicated overall good performance of commercial assays with genotype E samples (31,32,43) but also suggested that amino acid changes within and outside the "a" determinant may affect HBsAg recognition (37). The performance of the three reference assays in this study was overall consistent with that reported by previous studies using European or reference samples (27,32,43). However, we identified a significant impact of the T123A mutation on the performance of the Architect assay.…”

“…Genotype E shows approximately 8% amino acid divergence from genotype A, which is commonly used as the reference virus type for the development of HBsAg diagnostic assays (35). Previous studies indicated overall good performance of commercial assays with genotype E samples (31,32,43) but also suggested that amino acid changes within and outside the "a" determinant may affect HBsAg recognition (37). The performance of the three reference assays in this study was overall consistent with that reported by previous studies using European or reference samples (27,32,43).…”

Hepatitis B Surface Antigen (HBsAg) Assays—Are They Good Enough for Their Current Uses?

“…Diagnostic ("clinical") sensitivity may be related to the analytical performance of the assay used. For example, Scheiblauer et al evaluated 17 different diagnostic reagent sets available in Europe, and found that detection limits varied by 5-to 10-fold between the most sensitive and least sensitive assays, depending on the serotype of HBsAg tested (10 ).…”

Occult hepatitis B virus infection among Egyptian blood donors

“…Although, the incidence of transfusion-transmitted hepatitis B has been steadily reduced over the last four decades [3] , HBV still remains the most frequent transfusion-transmitted viral infection [4][5][6] . Being the first-line of blood screening for HBV [7] , different hepatitis B surface antigen (HBsAg) assays showed a range of sensitivity between < 0.1 and 0.62 ng of HBsAg per mL; 1 ng/mL corresponds to approximately 2 IU/mL [8,9] , However, there is clear evidence that transmission by HBsAgnegative components occurs, in part, during the serologically negative window period, but more so during the late stages of infection [1] , the later is referred to as occult HBV infection (OBI).…”