PCR-电化学基因芯片法在人乳头瘤病毒感染检测中的价值

Methodological evaluation of PCR-electrochemical gene chip in detecting human papillomavirus infection

目的对PCR-电化学基因芯片法检测人乳头瘤病毒(HPV)核酸进行方法学评价。方法收集贵州省某三甲医院309例宫颈脱落细胞有效样本,采用PCR-电化学基因芯片法检测人乳头瘤病毒并分型,以Sanger测序法为金标准,并与临床常用的荧光PCR法作比较,分别统计分析PCR-电化学基因芯片法与Sanger测序法和荧光PCR法检测结果的一致性和差异,并比较PCR-电化学基因芯片法和荧光PCR法的诊断效能。结果采用3种方法分别对309例有效样本进行HPV检测,均检测到20种不同的HPV基因型;PCR-电化学基因芯片法与Sanger测序法(Kappa=0.955,P<0.001)和荧光PCR法(Kappa=0.944,P<0.001)检测结果均有较高的一致性;与测序法金标准相比,PCR-电化学基因芯片法出现4例假阴性,1例分型结果不一致;PCR-电化学基因芯片法的敏感性为98.44%、特异性为100%、阳性预测值为100%、阴性预测值为92.86%、符合率为98.71%,荧光PCR法敏感性为100%、特异性为98.08%、阳性预测值为99.61%、阴性预测值为92.86%和符合率为99.68%。结论PCR-电化学基因芯片法与Sanger测序法和荧光PCR法的一致性良好,且具有高特异性和阳性预测值,可作为临床HPV感染检测方法的补充。

Objective To evaluate the methodological value of PCR-electrochemical gene chip in detecting human papillomavirus(HPV)gene.Methods Cervical exfoliated cells from 309 effective cases in a tertiary hospital in Guizhou Province were collected.PCR-electrochemical gene chip method,Sanger sequencing and fluorescent PCR commonly used in clinical practice were used to detect and type HPV.The consistency and differences in the results were compared and analyzed among PCR-electrochemical gene chip,Sanger sequencing,and fluorescent PCR.Diagnostic efficiency of PCR-electrochemical gene chip and fluorescent PCR were compared.Results Every method was able to detect 20 different HPV genotypes in 309 samples.The result from PCR-electrochemical gene chip method was highly consistent with those of Sanger sequencing method(Kappa=0.955,P<0.001)and fluorescent PCR method(Kappa=0.944,P<0.001).When compared to the gold standard of sequencing method,PCR-electrochemical gene chip method had 4 false negatives and 1 wrong typing.The sensitivity,specificity,positive predictive value,negative predictive value,and coincidence rate of PCR-electrochemical gene chip method were 98.44%,100%,100%,92.86%,and 98.71%,respectively,while those of fluorescent PCR method were 100%,98.08%,99.61%,92.86%,and 99.68%.Conclusion The results from PCR-electrochemical gene chip method have good consistency with those from Sanger sequencing method and fluorescent PCR method,and also have high specificity and positive predictive value.It can be used as a supplement to for clinical HPV infection detection methods.