Cambron Pierre, Jacquet Jeanne-Marie, Hoet Bernard, Lievens Marc
GlaxoSmithKline Biologicals, Rue de l'Institut 89, Rixensart, Belgium.
Clin Vaccine Immunol. 2009 Aug;16(8):1236-46. doi: 10.1128/CVI.00431-08. Epub 2009 Jun 24.
Pending removal from the market of a commercial assay (the AUSAB [Abbott Laboratories] enzyme immunoassay [EIA]) for the determination of antibodies to hepatitis B surface antigen (HBsAg), a new in-house quantitative enzyme-linked immunosorbent assay (ELISA) to measure antibodies against HBsAg (anti-HBs) was developed (anti-HBs in-house). Specific anti-HBs antibodies were sandwiched between the precoated HBsAg ad and ay subtypes purified from plasma from hepatitis B virus (HBV) human carriers and the recombinant HBsAg adw2 subtype tagged with horseradish peroxidase. The assay was calibrated against the 1st International Reference Preparation for anti-hepatitis B immunoglobulin (lot 1977-W1042). Analytical sensitivity and the limit of quantitation were estimated at 0.43 mIU/ml and 2.0 mIU/ml, respectively. Overall reproducibility was 11.86%, and accuracy was estimated to be 94.89%. More than 4,000 samples from seven clinical trials were tested with the anti-HBs in-house assay and compared to results generated with AUSAB EIA and AUSAB radioimmunoassay (RIA). During the technical validation, the anti-HBs in-house assay was compared to the AUSAB RIA as a reference (n = 919). Overall assessment of concordance and Deming's regression analysis were performed. The coefficient of correlation between the AUSAB RIA and anti-HBs in-house assay was 0.9815 with a slope of 0.9187. The overall agreement between anti-HBs in-house and AUSAB RIA was 97.61%, considering the clinical cutoffs at 3.3 mIU/ml and 1.0 mIU/ml for the respective assays. From a clinical perspective, seroprotection rates and anti-HBs geometric mean antibody concentrations for individual studies calculated with either the in-house assay or the reference assays were similar. Conclusions of individual studies were confirmed. The performance characteristics of the in-house assay are acceptable. There is no evidence that use of the new assay would lead to different clinical conclusions from the reference method.
在用于检测乙型肝炎表面抗原（HBsAg）抗体的商业检测方法（AUSAB[雅培实验室]酶免疫分析[EIA]）退市期间，开发了一种新的用于测量抗HBsAg抗体（抗-HBs）的内部定量酶联免疫吸附测定（ELISA）方法（抗-HBs内部检测法）。特异性抗-HBs抗体夹在预包被的从乙肝病毒（HBV）人类携带者血浆中纯化的HBsAg ad和ay亚型以及标记有辣根过氧化物酶的重组HBsAg adw2亚型之间。该检测方法以第一国际抗乙型肝炎免疫球蛋白参考制剂（批号1977-W1042）进行校准。分析灵敏度和定量限分别估计为0.43 mIU/ml和2.0 mIU/ml。总体重现性为11.86%，准确性估计为94.89%。来自7项临床试验的4000多个样本用抗-HBs内部检测法进行检测，并与AUSAB EIA和AUSAB放射免疫分析（RIA）产生的结果进行比较。在技术验证期间，将抗-HBs内部检测法与作为参考的AUSAB RIA进行比较（n = 919）。进行了一致性的总体评估和戴明回归分析。AUSAB RIA与抗-HBs内部检测法之间的相关系数为0.9815，斜率为0.9187。考虑到各自检测方法在3.3 mIU/ml和1.0 mIU/ml的临床临界值，抗-HBs内部检测法与AUSAB RIA之间的总体一致性为97.61%。从临床角度来看，用内部检测法或参考检测法计算的各研究的血清保护率和抗-HBs几何平均抗体浓度相似。各研究的结论得到了证实。内部检测法的性能特征是可接受的。没有证据表明使用新检测法会导致与参考方法不同的临床结论。
