乙肝泛变异HBsAg检测方法的建立及其在临床的初步应用  

Development of a method to detect immune escape mutant HBsAg of HBV widely and its initial use in clinic

作　　者：刘静华[1] 黄永国[1] 张春燕[1] 彭静[2] 张小燕[1] 龚劲松[1] 陈妍[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实验医学研究中心,湖北武汉430030 [2]华中科技大学同济医学院附属同济医院检验科,湖北武汉430030

出　　处：《中国实验诊断学》2008年第10期1199-1203,共5页Chinese Journal of Laboratory Diagnosis

基　　金：国家973计划项目(2005CB522901)资助

摘　　要：目的研制一种能全面检测血清野生及免疫逃逸变异HBsAg的ELISA试剂。方法采用抗HBs G6-mAb包被,羊抗HBs-HRP作示踪抗体,建立双抗体夹心ELISA。采用抗原替代试验,竞争抑制试验与其它多种试验一道对方法的特异性,敏感性与稳定性进行考核,并将其与市售HBsAg ELISA试剂一道用于一组HBV感染相关者的临床检测。结果证实本研究建立的ELISA具有很好的特异性与稳定性,其最低可检测浓度低于0.125 ng/ml。将其与两种市售HBsAgELISA用于乙肝病毒感染相关者检测,三法检测敏感性相当,其阳性率前者(34/177例,19.21%)分别高于或显著高于后两者(14.89%与6.21%,P分别<0.05及<0.01)。采用雅培化学发光试剂对部分本法阳性者进行复核,两法符合数为94.4%(17/18),两者间信号强度亦表现出一定的相关性。结论建立一个特异,敏感而稳定的血清HB-sAg检测ELISA,其对血清免疫逃逸变异HBsAg的检测能力超过国内市售ELISA。Objective Development of an ELISA, which can detect wide-type and immune escape mutant-type HBsAg wholly. Methods Results The sandwich ELISA was developed, which was based on the anfi-HBs G6 mAb and HRP-labeled goat anti-HBs antibody flint were used as coating and tracing antibody respectively. Using the substitution, competitive inhibition experiments and some other experiments to check the specificity, sensibility, and stability respectively. Meanwhile, put this method and other market ELISA into use in a group of clinical screening,which were related to HBV infection.thls ELISA possessed good specificity,sensibility,and stability. Its lowest screening concentration was less than 0.125 ng/ml.The sensibility of this method was correspond to other market ELISA in a group of clinical screening, the positive rate was 19.21% （34/177）, which was higher than the latter two. To check the positive that detected by this method through Abbott chemiluminescence kits. The coincidence rate was 94.4 % （ 17/18 ）. The sigruff intensity between the two also showed some correlation. Conclusion To design a specific, sensible and stable EIJSA, which screening capability of immune escape mutant HBsAg was higher than the domestic market ELISA.

关 键 词：乙型肝炎病毒 基因变异 乙型肝炎病毒表面抗原 免疫逃逸 酶联免疫吸附实验 

分 类 号：R446.61[医药卫生—诊断学]