A hook-effect-free homogeneous light-initiated chemiluminescence assay: is it reliable for screening and the quantification of the hepatitis B surface antigen?

Abstract

Background: Hepatitis B virus (HBV) infection remains a threat to global public health. As a hallmark of HBV infection, hepatitis B surface antigen (HBsAg) has been used to screen for HBV infection for decades, and quantitative assays are also being clinically rejuvenated to predict the disease outcome and monitor the antiviral response. Herein, we developed and evaluated a hook-effect-free homogeneous quantitative HBsAg assay based on the light-initiated chemiluminescence immunoassay (LICA).Methods: A hook-effect-free LICA algorithm was established by measuring the relative light units (RLUs) of two time points during the immunoreaction. The precision was assessed using low-and high-level controls. Consecutive clinical serum samples were tested using the LICA and Abbott Architect assay; samples producing inconsistent results were retested using supplementary assays including the HBsAg neutralization, HBV DNA, and Roche Elecsys HBsAg assays for further confirmation. The consistency, sensitivity, specificity, and positive and negative predictive values (PPV and NPV) were calculated. For the quantitative results, the correlation was analyzed. The coverage of different genotypes and mutations by the LICA was evaluated. Moreover, serial on-treatment and follow-up samples from chronic hepatitis B patients were also measured using the two assays.Results: The LICA had better within-run and within-laboratory precisions than the Architect assay. In total, 5,176 clinical samples were tested. The two assays showed a consistency of 99.63%. The LICA showed greater specificity (99.95% vs. 99.77%) and PPV (99.75% vs. 98.77%) than the Architect assay, whereas the Architect assay showed greater sensitivity (100.00% vs. 99.01%) and NPV (100.00% vs. 99.82%). The two assays displayed an excellent correlation independent of genotypes and mutations. The LICA hookfree algorithm recognized 100% of the underestimated results. Furthermore, similar HBsAg dynamics were demonstrated using the LICA and Architect HBsAg assay. Conclusions:The hook-free LICA provides a reliable tool for screening for HBV infection and quantifying HBsAg.

Correlation between Results of Semi-Quantitative and Quantitative Tests for Hepatitis B Virus Surface Antigen among Patients Achieving Viral Suppression with Antiviral Treatment

“…One-step sqHBsAg immunoassays provide faster results by completing the reaction in a homogeneous system without washing steps. However, the hook effect occurs when excess antigen binds to the capture and detection antibodies, preventing an immune response and leading to underestimated or false-negative results [ 27 ]. Yang et al have reported a hook effect-free qHBsAg chemiluminescence immunoassay, [ 27 ] and when we performed a stratified analysis by qHBsAg titer (≥ 1000 IU/mL) in HBeAg-positive CHB patients due to the hook effect, there was a negative correlation between qHBsAg and sqHBsAg.…”

Performance characteristics of the light-initiated chemiluminescent assay for quantitative determination of progesterone

“…Finally, the LICA is based on a one-step homogeneous immunoreaction, and the homogeneous environment does not requiring washing processes, which avoids the possibility of crosscontamination. Yang et al consider this an important factor responsible for the increased precision because it contributes to a more stable reaction system (12).…”

A new method for broadening the detection range of immunoassay and its application in β-hCG quantitative detection

“…In addition, the relative area of the chemibead is large, and antibodies with different affinities could be coated on its surface in different proportions. [24][25][26] If different chemibeads are mixed in different proportions, the local concentration of mAbs on the surface of the chemibeads and the collision probability of the chemibeads with b-hCG can be adjusted by the concentration of the beads. In this way, mAbs with different affinities could selectively play a role depending on the concentration of b-hCG in the sample, which ensures the analytical sensitivity and broadens the detection range.…”