A rapid and sensitive method based on magnetic beads for the detection of hepatitis B virus surface antigen in human serum

Abstract

Current clinically assays, such as enzyme-linked immunosorbent assay and chemiluminescence immunoassay, for hepatitis B surface antigen (HBsAg) are inferior in terms of either sensitivity and accuracy or rapid and high-throughput analysis. A novel assay based on magnetic beads and time-resolved fluoroimmunoassay was developed for the quantitative determination of HBsAg in human serum. HBsAg was captured using two types of anti-HBsAg monoclonal antibodies (B028, S015) immobilized on to magnetic beads and detected using europium-labeled anti-HBsAg polyclonal detection antibody. Finally, the assay yielded a high sensitivity (0.02 IU/mL) and a wide dynamic range (0.02-700 IU/mL) for HBsAg when performed under optimal conditions. Satisfactory accuracy, recovery and specificity were also demonstrated. The intra- and interassay coefficients of variation were 4.7-8.7% and 3.8-7.5%, respectively. The performance of this assay was further assessed against a well-established commercial chemiluminescence immunoassay kit with 399 clinical serum samples. It was revealed that the test results for the two methods were in good correlation (Y = 1.182X - 0.017, R = 0.989). In the current study, we demonstrated that this novel time-resolved fluoroimmunoassay could be used: as a highly sensitive, automated and high-throughput immunoassay for the diagnosis of acute or chronic hepatitis B virus infection; for the screening of blood or organ donors; and for the surveillance of persons at risk of acquiring or transmitting hepatitis B virus.

Simple and rapid monitoring of doxorubicin using streptavidin-modified microparticle-based time-resolved fluorescence immunoassay

“…Europium is one of the most frequently used lanthanide chelates and it was used here to label the competitive antigen. Magnetic microparticle-based TRFIA is a development of general TRFIA that has been reported for different applications such as cancer, 23,24 infectious disease, 25,26 and prenatal screening 27 and ts well in routine detection owing to advantages such as high precision, ultra-sensitivity, and its simple automated procedure. In addition, magnetic microparticles have high specic surface area and can be rapidly collected and separated from superuous analytes in a magnetic eld, shortening reaction time by increasing the uid exchange and automation of the washing and separation process.…”

Establishment of Magnetic Microparticles-Assisted Time-Resolved Fluoroimmunoassay for Determinating Biomarker Models in Human Serum

“…With the utilization of MMPs and Eu 3+ chelate, the MMPs-assisted TRFIA achieved high sensitivity and wider linear range. It is our belief that the novel technology platform combined with high speed and sensitivity seen in this and our previous studies [ 37 ] has great potential in rapid and high-throughput immunoassay testing and will bring about major improvements in the in vitro diagnostic and monitoring of patients with a wide range of clinical conditions.…”

Sequential injection-differential pulse voltammetric immunosensor for hepatitis B surface antigen using the modified screen-printed carbon electrode

“…Several methods have been reported for detection of HBsAg including enzyme-linked immunosorbent assay (ELISA) [2], chemiluminescent immunoassay [3], and fluoroimmunoassay [4]. Although these methods provide good sensitivity, precision, and selectivity, they are complicated, have long analysis time and high cost.…”