酶联免疫吸附试验临床检测信号临界区(灰区)患者血清HBsAg携带状况的研究  

Investigation on serum HBsAg-carrying conditions of patients whose serum HBsAg ELISA signals were in gray area

作　　者：张春燕[1] 龚劲松[1] 刘静华[1] 彭静[1] 黄永国[1] 张小燕[1] 李方和[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院,武汉430030

出　　处：《中国预防医学杂志》2009年第8期689-692,共4页Chinese Preventive Medicine

基　　金：国家十五科技攻关项目(2001BA705B05)

摘　　要：目的探讨乙肝HBsAg酶联免疫吸附试验（ELISA）检测信号灰区与血清免疫逃逸变异HBsAg之间的关系。方法采用市售ELISA对一组临床患者做常规检测,根据检测结果选留ELISA信号处灰区（即P/C 1.2~5.0之间）的实验标本。采用G6-ELISA（本室研制,有极强免疫逃逸变异HBsAg检测能力）与市售（基本不具备免疫逃逸变异HBsAg检测能力）对选留标本进行检测,以差减ELISA确认其免疫逃逸变异HBsAg阳性数量,并采用雅培-化学发光及PCR对检测结果的特异性进行复核。结果自10 231例受检者中筛选HBsAg ELISA信号灰区标本244例。以G6-ELISA与市售ELISA检测结果并做差减分析显示免疫逃逸变异HBsAg阳性率为11.07%（27/244）,亚临界,临界及超临界各组阳性率分别为9.30%（8/86）,10.34%（15/145）及30.77%（4/13）,后者组阳性率显著高于前两组（P〈0.01）。考核结果显示,经ELISA差减分析确认的免疫逃逸变异HBsAg阳性血清雅培-化学发光检测阳性率为94.44%（17/18）,HBV DNA阳性率为33.33%（6/18）,其中两份单项G6-ELISA检测最低阳性信号标本的HBsAg特异性亦为雅培-化学发光检测所证实。结论临床HBsAg检测样本中存在少量A（或P/C）值在ELISA信号灰区的异质化标本;免疫逃逸变异HBV感染是造成这一现象的主要原因之一。Objective To discuss the relationship between the signal gray area formed by HBsAg ELISA detection and immune escape mutant HBsAg （ IEM HBsAg）. Methods Market ELISA A kits were used to detect a group of clinical patients＇ sera as usual, then those serum samples whose ELISA signals were in gray area （P/C between 1.2-3.0） according to the results were selected. Self-made G6-ELISA and market ELISA B kits, which had great and scarce reaction to IEM HBsAg respectively, were used to detect the selections. The difference between the two kinds of ELISA （ differential ELISA） was used to confirm the positive numbers of the immune escape mutant HBsAgs, then the partial serum samples detected by differential ELISA were checked by PCR and Abbott immunochemiluminescent kits. Results There were 244 gray area examples screened from 10 231 examples using market ELISA A. According to the results of the differential analysis which consists of G6 ELISA and market ELISA B, the positive rate of IEM HBsAg was 11.07% （27/244）, and the positive rates of subcritical, critical and the supercritical groups were 9. 30% （8/86）, 10. 34% （15/145） and 30. 77% （4/13） , respectively, and the positive rate was remarkably higher in the supercritical group than in the first two groups （P 〈0.01 ）. The checking results showed that, in those IEM HBsAg positive serum samples detected by differential analysis, the positive rate was 94.44% （17/18） by Abbott immunochemiluminescent kits, and the positive rate was 33. 33% （6/t8） by HBV DNA sequencing. The two HBsAg specific samples, which showed the least positive signal detected by G6-ELISA, were also verified by Abbott immunochemiluminescent kits. Conclusion There are a small amount of serum samples in clinical samples whose A （ P/C ） values stay in the gray area by HBsAg ELISA, and the immune escape mutant HBV infection is one of the main reasons causing this phenomenon.

关 键 词：乙型肝炎病毒 基因变异 乙型肝炎表面抗原 免疫逃逸变异 酶联免疫吸附试验 

分 类 号：R446.61[医药卫生—诊断学]