Objective: To evaluate and compare the detection consistency of hepatitis B surface antigen (HBsAg) by two immunoassays: enzyme-linked immunosorbent assay (ELISA) and electrochemiluminescent immunoassay (ECLIA). Methods: A prospective study was conducted among 2 296 pregnant women recruited consecutively from January 1, 2014 to January 31, 2015 in a hospital. Blood samples were collected from them for the detection of HBsAg by using ELISA and ECLIA, Kappa test was performed on the results. Nested polymerase chain reaction and sequencing of HBV S gene were also performed in all samples. Phylogenetic analysis was performed using Mega 6.0 software. Results: The two methods had high detection consistence of HBsAg (Kappa=0.71). There were significant differences in detection result of B genotype and adw2 serotype HBV strains between two methods. Among 123 identified HBV strains, 113 belonged to genotype B and available for further analysis. The difference in detection of substitution rates between two methods or different positive groups were not significant. Compared with ELISA single positive group, the ECLIA single positive group had completely different substitution sites. Conclusion: The two methods had high detection consistence of HBsAg, but there were still 32.4% HBV DNA positive cases in ELISA/ECLIA single positive group, and complete complementary substitution sites between ELISA single positive group and ECLIA single positive group. Our results suggested that more effective detection procedure should be considered for the possible impact of the HBV silent transmission and infection.
目的： 比较ELISA和电化学发光免疫法（ECLIA）检测HBsAg的一致性，探讨其相关的因素和基因进化特征。 方法： 2014年1月1日至2015年1月31日连续招募安庆市市立医院产科初次住院活产孕妇2 296例，收集其人口学特征，并对采集的血清应用ELISA和ECLIA检测HBsAg，采用Kappa检验评价两种方法检测结果的一致性；对HBV S基因片段进行巢式PCR扩增、测序及进化分析。 结果： 两种方法具有一致性，Kappa=0.71。共获得123株HBV S基因片段序列，其中113株为B基因型（adw2血清型112株，adw3血清型1株），10株为C基因型（全为adr血清型）。两种方法检测不同基因型或血清型病毒株的结果显示，ECLIA对B基因型和adw2血清的检测明显优于ELISA，差异有统计学意义。113株B基因型序列进化分析显示，两种检测方法或不同阳性组别在各位点替换率的差异均无统计学意义。发生于HBsAg主要亲水区域（MHR）的氨基酸变异显示，ELISA/ECLIA单纯阳性组出现完全不相同的替换位点。 结论： 两种方法检测HBsAg具有高度的一致性，但在ELISA/ECLIA单纯阳性组中仍获得34株（32.4%）HBV S基因片段，且存在MHR的氨基酸替换位点互补。因此在控制HBV传播中应考虑ELISA/ECLIA单纯阳性的感染者在人群中可能的静默传播作用，还需优化临床检测程序。.
Keywords: Electrical chemiluminescent immunoassay; Enzyme-linked immunosorbent assay; Hepatitis B surface antigen.