【摘要】 目的建立巢式PCR(nPCR)检测人乳头状瘤病毒的方法学及作临床应用。方法以MY09/MY11为外引物,GP5+/GP6+为内引物建立nPCR,优化体系,评价其特异性和灵敏度,同时检测了54例HPV DNA阳性、4例弱阳性、42例阴性的标本,与基因导流杂交法作一致比较。结果检测到单一目的片段;最低能检测到1∶16 384稀释的HPV18阳性对照品,明显高于MY09/MY11方法;nPCR法检测到60例HPV DNA阳性标本,40例HPV DNA阴性的标本,与基因导流杂交法具有较好的一致性(χ2=39.518,P<0.001;Kappa=0.628,P<0.001)。结论所建立的nPCR具有较好的特异性和灵敏度,与基因导流杂交法具有较好一致性。更多还原

【Abstract】 OBJECTIVE To establish a nested-PCR(nPCR) method to detect the human papillomavirus and analyzed its preliminary clinical practice.METHODS MY09/MY11 was as outer primer and GP5+/GP6+ as inner primer to establish a nPCR and optimiz the PCR reaction system,and its specificity and sensitivity were appraised.The 54 HPV positive samples,4 weak positive samples,and 42 negative samples were simultaneously detected by nPCR and compared with the flow-through hybridization.RESULTS The established nPCR method could detect the sole goal segment and 1∶16 384 dilution HPV18 positive control,so it was more obviously sensitive than the MY09/MY11.The nPCR method detected 60 HPV positive samples and 40 negative samples,which had the good uniformity with the flow-through hybridization(χ2=39.518,P<0.001,Kappa=0.628,P<0.001).CONCLUSIONS The nPCR has the good specificity,sensitivity and uniformity with the flow-through hybridization.更多还原