HBV Surface Antigen (subtype adw2, mutation M133L)

Background: Hepatitis B surface antigen (HBsAg) remains the main viral marker for screening and monitoring hepatitis B virus (HBV) infection. The quantification limit of most current HBsAg assays is around 0.05 IU/mL. The Lumipulse-G-HBsAg-Quant assay (Fujirebio) claims to obtain a tenfold improvement in sensitivity. This study aimed to assess the performance of this assay in detecting low HBsAg levels in clinical samples. Methods: Three panels of stored frozen samples were selected on the basis of HBV-DNA and HBsAg values obtained previously with routine techniques. Panels 1 (n=13) and 2 (n=52) consisted of DNA-positive/HBsAg-negative samples from individuals in the window period and with occult HBV infection respectively. Panel 3 comprised 23 samples with low or discrepant HBsAg screening results. All these samples were tested retrospectively with the DiaSorin and Fujirebio HBsAg assays. Results: Sixteen out of 65 samples (25 %), initially screened HBsAg negative, were reactive only with the Fujirebio assay (median value= 0.015 IU/mL; IQR= 0.012): three (23 %) samples from panel 1 and 13 (25 %) from panel 2. Thirteen of these 16 (81 %) had HBsAg values below 0.03 IU/mL with the DiaSorin assay. In panel 3, 22 (96 %) samples were quantified successfully with the Fujirebio assay (median: 0.32 IU/mL; IQR: 1.20) and 19 (83 %) with the DiaSorin assay (median: 0.31 IU/mL; IQR: 0.65). Concentrations obtained with the two assays showed good correlations (r=0.893, Spearman). Conclusions: HBsAg assays with enhanced analytical sensitivity could improve HBV serological profile interpretation with possible consequences on clinical management of infected patients, and on blood transfusion safety.

Herein, a novel sandwich-type electrochemical immunosensor was fabricated based on Fe3O4 nanoflowers (Fe3O4 NFs) and heterogeneous chain reaction (HCR) signal amplification strategy for the sensitive detection of hepatitis B surface antigen (HBsAg). The aldehyde-functionalized Fe3O4 NFs are used as a supporting matrix to immobilize the hepatitis B surface antibody 1 (HBsAb1). The biotin-modified single-strand DNA (biotin-SO) was connected onto the biotin-HBsAb2 via linkage of streptavidin (SA), followed by addition of methylene blue (MB) modified single strand DNA1 (MB-S1) and DNA2 (MB-S2) for HCR signal amplification. The designed immunosensor exhibited a detection linear range of 0.5 pg mL(-1)-0.25 ng mL(-1) and a low detection limit of 0.16 pg mL(-1), with excellent stability, selectivity and reproducibility. Furthermore, HBsAg is detected in the serum samples with a stable and fast response, indicating that the proposed immunosensor has a promising potential application in clinical analysis.