液相芯片技术检测乙型肝炎病毒表面抗原

目的建立乙型肝炎病毒表面抗原(HBsAg)液相芯片检测法并进行评估。方法将4种不同来源的HBsAg单克隆抗体分别包被在不同的羧基化微球上,利用双抗体夹心法原理建立液相芯片检测HBsAg的方法。检测定值血清、标准血清盘以及本中心献血者筛选保留的随机43份HBsAg阳性标本和500份HBsAg阴性标本,并评估方法的敏感性和特异性。结果液相芯片法检测HBsAg灵敏度可达到0.2IU/ml。在标准血清盘中,HBsAg阳性符合率为25/25,阴性符合率为14/15。在收集的43份阳性标本、500份阴性标本中,HBsAg阳性标本符合率为100%,阴性标本符合率分别为99.4%。4种不同来源HBsAg单克隆抗体的检测结果存在差异,可以互补。结论建立的液相芯片方法可有效检测HBsAg。

Establishment of multiplexed Luminex assay for detection of hepatitis B surface antigen

Objective To establish a multiplexed Luminex assay for detection of hepatitis B surface antigen (HBsAg) and to evaluate the sensitivity and specificity. Methods Four HBsAg monoclonal antibodies from different manufactures were coupled to the different Luminex carboxylated fluorescent microspheres. A multiplexed Luminex assay was established to analyze HBsAg based on double-antibody sandwich principle. The panel samples and known standard samples were detected for validating sensitivity and specificity of the assay. Results The detection limit of HBsAg was 0.2 IU/ml by the multiplexed Luminex assay. The positive consistent rate and negative consistent rate were 25/25 and 14/15 in the HBsAg panel samples. The positive and negative consistent rates were 100% and 99.4% respectively, according to test results of 43 positive and 500 negative samples. The results of samples using four different antibodies were different and mutually complementary. Conclusion The multiplexed Luminex assay was reliable and could detect HBsAg effectively.