金标免疫层析法测定唾液中丙型肝炎病毒抗体的研究

Study on anti HCV in saliva by colloid gold-based immunochromatographic assay

目的研制一种能快速检测唾液中丙肝IgG、IgM抗体的胶体金免疫层析试纸条。方法采用夹心法,将丙肝重组抗原与胶体金链接形成金标抗原,与金标兔抗体以2∶1的比例混合制成金标垫。将鼠抗人IgG、鼠抗人IgM抗体混合包被于检测线位置;将羊抗兔抗体置于质控线位置;在样品垫上滴加10μL唾液和1滴PBS溶液(约45μL)展开。并对试纸条的特异性,重复性等进行测试。结果对217份血清抗-HCV阳性样品和189份阴性样品进行了测试,与血清抗-HCV ELISA检测方法的一致性高(Kappa=0.99)。结论本技术能在无创条件下,5min时间内可读取结果,可用于体检大量筛查和群众的自主筛查而推广。

Objective To triturate a gold immunochromatography test strip for detecting anti-Hepatitis C Virus（HCV） in saliva.Methods Colloid-gold was prepared according to the sodium citrate method,and then conjugated to HCV recombination antigen and rabbit antibody.Mouse anti-human IgG and IgM mixture antibodies were coated on a porous nitrocellulose membrane as the detection line and goat anti-rabbit IgG was coated as the control line.Results The developed colloid-gold immunochromatography assay was used to detect 217 positive samples and 189 negative samples,and kappa factor between serum ELISA and new technology was 0.99.And the developed technology exhibited an excellent stability and specificity.Conclusion A new colloid-gold immunochromatography strip for the detection of anti-HCV in saliva has been successfully developed.This method is sensitive,rapid（the detection procedure can be completed in 5 min） and suitable for crowed screening.