丙型肝炎病毒核酸液相芯片分型检测方法的建立及应用

Establishment and application of HCV genotype liquichip detection method

目的建立丙型肝炎病毒1a、1b、2a、3a、3b、6a亚型的核酸液相芯片分型检测方法。方法建立PCR扩增及核酸探针偶联方法,将PCR产物与偶联核酸探针的微球混合物进行杂交,建立液相芯片检测方法,并对建立的液相芯片检测方法进行灵敏度及特异性评价,应用该方法对93份血清样本核酸进行检测。结果建立的丙型肝炎病毒核酸液相芯片分型检测方法具有较高的特异性和敏感性,能对6种亚型进行检测和分型,对于HCV 1a、3a和6a亚型核酸液相芯片分型检测方法的灵敏度为1×105copies/PCR;对于HCV-1b、2a和3b亚型核酸液相芯片分型检测方法的灵敏度为1×104 copies/PCR。对93份临床样本的检测结果表明,该方法具有高通量、快速、敏感、特异的特点。结论本方法能对丙型肝炎病毒的6种亚型进行同时快速检测,为丙型肝炎病毒的分型检测提供一种新方法。

Objective To establish a liquichip method for detecting 6 sub-genotypes of hepatitis C virus（HCV）,including 1a, 1b,2a,3a,3b and 6a.Methods The coupling method of PCR amplification and nucleic acid probe was established.The PCR product and the microspheres mixture of the coupled nucleic acid probe were hybridized for establishing the liquichip detection method.The sensitivity and specificity of the established liquichip detection method were evaluated.Nucleic acid in 93 serum samples was detec-ted by this method..Results The established HCV nuclei acid liquichip genotype detection method had the higher specificity and sensitivity,which could detect and classfy 6 HCV sub-genotypes.The sensitivity for HCV 1a,3a and 6a sub-genotypes was 1× 105 copies/PCR;the sensitivity for HCV 1b,2a and 3b sub-genotypes was 1×104 copies/PCR.The detection results in 93 serum samples showed that the this genotyping method had the characteristics of high throughput,rapidness,sentsitivity and specificity. Conclusion This method can be used for the simultaneous and quick detection of 6 HCV sub-genotypes and provides a new meth-od for the genotyping detection of HCV.