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.
出处
《中国生物制品学杂志》
CAS
CSCD
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).
