用重组结构区抗原检测抗丙型肝炎病毒IgM抗体方法的建立

Establishment of a method for detecting anti-HCV IgM using recombinated structural region antigens

目的检测抗丙型肝炎病毒（ＨＣＶ）结构区蛋白ＩｇＭ抗体。方法采用丙型肝炎病毒Ｃ，Ｅ１，Ｅ２区重组抗原混合包被和分别包被酶标板；用兔抗人γ链血清处理人血清标本，再用固相包被羊抗兔抗体吸附兔抗人γ链－人ＩｇＧ复合物，建立了抗－ＨＣＶＩｇＭ检测方法。结果对７６例丙型肝炎病人血清进行抗－ＨＣＶＩｇＭ检测，同时与逆转录－巢式聚合酶链反应（ＲＴ－ＰＣＲ）检测结果进行比较，两者具有相关性（Ｐ＜０００５）；ＩｇＭ抗体组成分析结果表明采用全片段Ｃ、Ｃ＋Ｅ２区抗原血清标本中的抗－ＨＣＶＩｇＭ检出率分别可达９６６％和１００％。结论ＨＣＶ的Ｃ、Ｅ２重组抗原用于抗－ＨＣＶＩｇＭ检测具有较高的敏感度和特异性。

Presence of IgM antibody to viral structurale protein can be taken as the marker of its rdplication,so the detection method for anti-HCV IgM was established using as HCV recombinated structural region antigens,including C,E1,E2,as the coated antigen and rabbit anti-human γ chain sera for neutralization and sheep anti-rabbit IgG coated for the sorbence of the complex of RAH-γ with human IgG.76 serum specimens of the hepatitis C patients were detected by indirect ELISA for anti-HCV IgM and by RT-PCR for RNA. The results suggested that IgM antibodies to C,E1,E2 were closely related with RNA (P<0 005).The components of the IgM antibody,too were identified by indirect ELISA with recombinated antigens coated respectively.When whole-fragment C or C+E2 were used as the coated antigens, anti-HCV IgM in 96.6% or 100% of serum specimens can be respectively detected.Using recombinated structurae region antigens,detection of anti-HCV IgM by ELISA was high sensitivity and specificity.