采用PCR-测序法对北京地区325例妇女宫颈脱落细胞样品中人乳头瘤病毒进行检测和基因分型

PCR-based sequencing for detection and genotyping of human papillomavirus in 325 cervical cell specimens from Beijing area

摘要:

为了探讨PCR-测序法在宫颈脱落细胞样品中人乳头瘤病毒 (Human papillomavirus, HPV) 临床检测中的应用价值，采用HPV通用引物PGMY09/11针对HPV L1区基因序列进行PCR扩增，并通过DNA测序法对HPV进行基因分型。对于混合感染样品，利用HPV型别特异性引物PCR的方法进行基因分型。325例临床样品中，228例为HPV阳性，其中66例为混合感染。共发现27种不同的HPV型别，其中HPV 16比例最多，其次是HPV 58和52。高危型HPV检出率随病变程度加重显著性增加

Abstract:

To evaluate PCR-sequencing for clinical detection of human papillomavirus (HPV) genotypes in cervical cell specimens, we applied PCR-sequencing to HPV detection and genotyping by general primer PGMY09/11, which targets the HPV most conserved L1 gene. Samples with multiple infections were subjected to HPV type-specific PCR. Among the 325 cervical samples, 228 were HPV positive, of which 66 showed multiple infections. In all, 27 different HPV genotypes were identified, with HPV 16 being the most prevalent, followed by HPV 58 and 52. The prevalence of high-risk HPV infection increased with the severity of cervical lesions (P<0.05), whereas the proportion of multiple infections declined significantly from LSIL to SCC (P<0.05). Both rates of overall and high-risk HPV infection were the highest in 21-30 age groups. There was substantial agreement between the HC2 and PCR-sequencing assay for detection of high-risk HPV (kappa=0.675). PCR-sequencing was effective in HPV detection and genotyping, and it could be potentially applied to large scale HPV screening.