抗-HCV多表位抗体在检测HCV抗原中的应用  

Application of HCV Multi-epitope Antibody in Test for HCV Antigen

作　　者：谢忠平[1] 龙润乡[1] 宋霞[1] 李华[1] 李文忠[1] 熊秋霞[1] 洪超[1] 崔萍芳[1] 李琦涵[1] 

机构地区：[1]中国医学科学院中国协和医科大学医学生物学研究所质量检定室,昆明650118

出　　处：《中国生物制品学杂志》2006年第4期411-413,共3页Chinese Journal of Biologicals

基　　金：云南省人才基金资助(编号:2002PY11).

摘　　要：目的 应用丙型肝炎病毒多表位抗体，探索从血浆或血清样品中检测丙型肝炎病毒抗原（HCAg）的可能性，为献血员筛选及临床早期诊断提供检测方法。方法 利用原核系统表达的丙型肝炎病毒（HCV）7个高度保守区基因的多表位复合抗原免疫动物，制备抗-HCV多克隆抗体，采用ELISA双抗体夹心法检测血浆或血清中的HCAg，并与RT-PCR法进行比较。结果 制备的抗-HCV多表位复合抗原多克隆抗体，在用ELISA法检测HCAg中表现出较高的特异性及灵敏性，Cutoff值为0．239，批内CV值为5．60％～6．65％，批间CV值为7．80％。与RT-PCR比较，相关系数为0．816，灵敏度为88．89％，特异度为96．30％，一致性为95．24％，调整一致性为90．82％，阳性预测值为80．00％，阴性预测值为98．11％。HCAg检出率与美国ORTHO公司HCV—C抗原检测试剂盒差异无显著意义（P=0．06）。结论 用HCV多表位复合抗原制备的多克隆抗体，采用ELISA双抗体夹心法检测HCAg，具有灵敏、特异、快速的优点，有望开发成为HCAg诊断试剂盒。Objective To explore the possibility of test for hepatitis C virus antigen（HCAg） in plasma or serum samples by using HCV multi-epitope antibody and provide a method for screening of blood donors as well as early diagnosis of hepatitis C. Methods Immunize animals with the multi-epitope complex antigen of HCV expressed in E. coli to prepare HCV polyclonal antibody. Determine the HCAg in plasma or serum samples by double-antibody sandwich ELISA using the prepared polyclonal antibody and compare the determination result with that by RT-PCR. Results The prepared HCV polyclonal antibody showed high spoeificity（96. 30% ） and sensitivity（ 88. 89% ） in test for HCAg by ELISA. The cutoff value was defined as 0. 239. The intra- and inter-variation coeffeients （CV） were 5.60% -6. 65% and 7. 80% respectively. The correlation coefficient of ELISA and RT-PCR was 0. 816 ,and their consistency and adjusted consistency were 95.24% and 90. 82% respectively. The positive and negative prediction values of ELISA were 80.00% and 98.11% respectively. The detectable rates of HCAg by ELISA using the prepared HCV polyclonal antibody showed no significant difference from that using HCV-C antigen kit manufactured by ORTHO （ P = 0. 06）. Conclusion The prepared HCV polyelonal antibody was sensitive,specific and rapid in determination of HCAg by double antibody sandwich ELISA and might be used for the development of HCAg diagnostic kit.

关 键 词：丙型肝炎 多表位抗体 ELISA 

分 类 号：R720.597[医药卫生—急诊医学] R392-33[医药卫生—儿科]