摘要
目的制备抗乙型肝炎表面抗原(HBsAg)单克隆抗体,建立检测变异HBsAg的方法。方法用血源HB-sAg免疫BALB/c小鼠,制备可以识别变异HBsAg的抗-HBs单抗。筛选出可以较好识别变异HBsAg的单抗mAb1,优化该抗体ELISA检测HBsAg的方法,与2种HBsAg检测试剂比较检测变异HBsAg的能力。对6份临床样品进行检测及序列分析,以证实方法的可靠性。结果经过筛选,得到1种可以较好识别包括G145R在内的大多数变异HB-sAg的单抗。检测变异HBsAg的能力优于市售HBsAg诊断试剂。结论制备的单抗可以用于ELISA检测变异HB-sAg,减少HBsAg变异株的漏检率。
Objective To develop enzyme-linked immunosorbent assay(ELISA) of detecting hepatitis B surface antigen mutants using monoclonal antibody.Methods BABL/c female mice were immunized with HBsAg isolated from serum of HBV infected patients.All monoclonal antibodies were produced according to the standard methods.The reactivity of anti-HBsAg monoclonal antibodies named mAb1 to 13 different HBsAg variants was measured by using enzyme immunoassay.The immunoreactivity to mutant antigens was also tested using 8 commercial HBsAg diagnostic assays.Six clinical samples were detected using mAb1 and the sequences of the 6 samples were analyzed.Results One anti-HBsAg monoclonal antibody(mAb1) reacted strongly with most tested variants including variant with G145R mutation.The ability of detected HBsAg variants was better than commercial HBsAg diagnostic assays.Conclusion The mAb1 is a valuable tool for HBsAg variants detection.
出处
《华中科技大学学报(医学版)》
CAS
CSCD
北大核心
2007年第5期667-670,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
湖北省科技攻关计划资助项目(No2006AA301B51-1)
关键词
乙型肝炎表面抗原
单克隆抗体
酶联免疫吸附测定
hepatitis B surface antigen
monoclonal antibody
enzyme-linked immunosorbent assay