解放军医学杂志2017，Vol.42Issue(10)：865-869,5.DOI:10.11855/j.issn.0577-7402.2017.10.05
隐匿性HBV感染相关S基因突变对HBsAg检测的影响及其机制
Analysis of the influence of nine occult HBV infection-related S-gene mutations on HBsAg detection
摘要
Abstract
Objective To determine the reactivity of occult HBV infection (OBI)-related S-gene mutations to various anti-HBs antibodies (Abs) and HBsAg detection reagents.Methods Nine representative S-gene mutations M1-M9 (including 5 novel mutations M1-M5) from 1 OBI patient and 3 OBI blood donors were investigated.S-gene recombinant plasmids harboring 9 mutants or wild-type sequences constructed before were transfected into Chinese hamster ovary (CHO) cells,respectively.HBsAg expression levels and reactivity with various anti-HBs Abs and regular HBsAg detection reagents of all the mutants were analyzed.Results Compared to wild-type strain,intracellular HBsAg levels of the 9 mutants were obviously reduced upon Roche quantitative Elecsys assay.In contrast,analysis of the same samples using anti-His-tag Ab showed that the levels of HBsAg-Histag fusion protein were significantly reduced only in mutants M1,M6,and M7.The results of reactivity of mutant HBsAg against 6 anti-HBs Abs (S/CO values) showed that poor reactivity was observed for most mutants.Specifically,M1-,M4-,M7-,and M9-produced HBsAg with Ab4,and M9-produced HBsAg with Ab6 had the worst reactivity (S/CO＜1) compared to the wild-type.The results of reactivity of mutant HBsAg with 6 commercial HBsAg detection reagents showed that the reactivity for most mutants was significantly lower than that of the wild-type (P＜0.05).The miss rates of ELISA reagents D,E,and F were 11.1％,22.2％,and 55.6％,respectively.Conclusions Lower-affinity of studied mutant HBsAg with anti-HBs is one of the major causes of OBI presentation.The regular HBsAg detection reagents used currently in clinic show a significant deficiency in detection of the mutant HBsAg and thus need to be improved.关键词
乙型肝炎病毒/S基因/突变/表面抗原Key words
hepatitis B virus/S gene/mutation/HBsAg分类
医药卫生引用本文复制引用
张凯,刘妍,陈容娟,李奇,许智慧,思兰兰,徐东平,蔺淑梅..隐匿性HBV感染相关S基因突变对HBsAg检测的影响及其机制[J].解放军医学杂志,2017,42(10):865-869,5.基金项目
国家自然科学基金(81572010,81373136) （81572010,81373136）
首都卫生发展科研专项项目(2016-2-5032) （2016-2-5032）
北京市自然科学基金(7172206).This work was supported by the National Natural Science Foundation of China (81572010,81373136),the Capital Health Research and Development of Special Project (2016-2-5032),and the Natural Science Foundation of Beijing (7172206) （7172206）
