Dual-Labeled Time-Resolved Immunofluorometric Assay for the Simultaneous Quantitative Detection of Hepatitis B Virus Antigens in Human Serum

Abstract

In this paper, a novel time-resolved fluorescence immunoassay (TRFIA) is described that allows the simultaneous quantitative detection of hepatitis B virus surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) in human serum to aid the diagnosis and monitoring of hepatitis B virus infection. The proposed method was developed based on a two-step sandwich immunoassay protocol in which monoclonal antibodies against HBsAg and HBeAg were co-coated in 96 microtitration wells, then tracer polyclonal antibodies against HBsAg labeled with samarium and tracer monoclonal antibodies against HBeAg labeled with europium chelates were used for detection. The detection range was 0.1-150 IU/mL for HBsAg and 0.5-160 PEIU/mL for HBeAg, and the detection limits were 0.03 IU/L and 0.09 PEIU/ml, respectively. The intra- and inter-assay coefficients of variation were below 8 % for both virus antigens. The dilution linearity and accuracy of the assay were satisfactory. No statistically significant differences were observed in sensitivity or specificity for the serum samples between the dual-label TRFIA and a commercial single-label TRFIA. These results demonstrate that an effective, reliable and convenient HBsAg/HBeAg dual-label TRFIA was successfully developed that may be clinically applicable for blood screening to monitor the course of hepatitis B virus infection and predict treatment responses.

Eu3+/Sm3+ dual‐label time‐resolved fluoroimmunoassay for measurement of hepatitis C virus antibodies

“…For example, the use of FQ‐PCR is limited by its complexity, and ELISA is only able to detect single antibodies. TRFIA is a novel technology that uses lanthanide elements for labeling, and it provides various benefits, such as a high sensitivity, wide linear range, and good tracer stability; in addition, it is not easily affected by sample autofluorescence . Moreover, multiple labeling analyses can be performed as the detection windows and wavelengths of lanthanide elements vary.…”

A New Method for Detection African Swine Fever Virus: Time-resolved Fluorescence Immunoassay

“…TRFIA is a sophisticated detection technique, and has been widely used in the detection of multiple viruses or their antigens [ 21 , 22 ]. Compared with ELISA, TRFIA has higher sensitivity and wider measurement range.…”