一种无钩状效应的均相光引发化学发光测定法：它对乙肝表面抗原的筛查和定量是否可靠？
A hook-effect-free homogeneous light-initiated chemiluminescence assay: is it reliable for screening and the quantification of the hepatitis B surface antigen?
作者信息
Yang Ruifeng, Cui Liyan, Liu Yan, Cong Xu, Fei Ran, Wu Shuping, Wei Lai
机构信息
Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing International Cooperation Base for Science and Technology on NAFLD Diagnosis, Beijing 100044, China.
Department of Clinical Laboratory, Peking University Third Hospital, Beijing 100191, China.
出版信息
Ann Transl Med. 2020 May;8(9):606. doi: 10.21037/atm.2020.02.59.
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% . 99.77%) and PPV (99.75% . 98.77%) than the Architect assay, whereas the Architect assay showed greater sensitivity (100.00% . 99.01%) and NPV (100.00% . 99.82%). The two assays displayed an excellent correlation independent of genotypes and mutations. The LICA hook-free 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.
背景
乙型肝炎病毒（HBV）感染仍然是全球公共卫生面临的威胁。作为HBV感染的标志，乙肝表面抗原（HBsAg）已被用于筛查HBV感染数十年，并且定量检测在临床上也正在复兴，以预测疾病转归和监测抗病毒反应。在此，我们开发并评估了一种基于光引发化学发光免疫分析（LICA）的无钩效应均相定量HBsAg检测方法。
方法
通过测量免疫反应过程中两个时间点的相对光单位（RLU），建立了一种无钩效应的LICA算法。使用低水平和高水平对照评估精密度。使用LICA和雅培Architect检测方法对连续的临床血清样本进行检测；对产生不一致结果的样本，使用包括HBsAg中和、HBV DNA和罗氏Elecsys HBsAg检测在内的补充检测方法进行重新检测，以进一步确认。计算一致性、敏感性、特异性以及阳性和阴性预测值（PPV和NPV）。对于定量结果，分析相关性。评估LICA对不同基因型和突变的覆盖情况。此外，还使用这两种检测方法对慢性乙型肝炎患者的系列治疗中和随访样本进行了检测。
结果
LICA的批内和实验室内精密度均优于Architect检测方法。总共检测了5176份临床样本。两种检测方法的一致性为99.63%。与Architect检测方法相比，LICA显示出更高的特异性（99.95%＞99.77%）和PPV（99.75%＞98.77%），而Architect检测方法显示出更高的敏感性（100.00%＞99.01%）和NPV（100.00%＞99.82%）。两种检测方法显示出与基因型和突变无关的良好相关性。LICA无钩算法识别出100%被低估的结果。此外，使用LICA和Architect HBsAg检测方法显示出相似的HBsAg动态变化。
结论
无钩LICA为筛查HBV感染和定量HBsAg提供了一种可靠的工具。
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