安顺地区受检妇女中高危型人乳头瘤状病毒的检出及单核苷酸多态性特征

Detection of high-risk human papillomavirus and characteristics of single nucleotide polymorphism in Anshun women

目的 探讨安顺地区受检妇女中高危型人乳头瘤病毒(HR-HPV)的检出及单核苷酸基因多态性(SNP)特征。方法 采集10 774例行HPV分型检测妇女的宫颈脱落细胞为细胞标本来源,选取其中HR-HPV阳性且有组织病理学活检指征的298例受检者的宫颈组织为组织标本来源,采用聚合酶链式反应(PCR)体外扩增和反向点杂交相结合检测宫颈脱落细胞的HPV基因分型,进一步采用SnapGene、MEGA11分析HPV52、HPV58型的SNP,采用苏木精-伊红(HE)染色判定宫颈组织的宫颈病变级别。结果 受检妇女中HR-HPV以单一感染为主、阳性率为20.36%,HR-HPV感染亚型主要为HPV52、HPV16、HPV58、HPV53及HPV51;宫颈病变中HR-HPV优势型别为HPV16、HPV52、HPV58、HPV18及HPV33;SNP和进化树分析发现,HPV52型E6和E7区SNP点分别有7个(其中A125T、A294G是新的突变位点,G350T、A379G突变率最高)和3个(其中T666C是新的突变位点,C751T、A801G突变率最高)、且HPV 52型均分布于B谱系,HPV58型E6、E7区SNP点分别有3个和8个、突变位点最高的是T744G、且HPV58型均分布于A谱系。结论 安顺地区受检妇女主要HPV阳性型别为HPV52、HPV16及HPV58型,宫颈病变中主要优势型别为HPV16、HPV52及HPV58型,且HPV52、HPV58有其特定的SNP。

Objective To explore the detection of high-risk human papillomavirus(HR-HPV)and single nucleotide gene polymorphism(SNP)in the women tested in Anshun area.Methods Cervical exfoliated cells from 10,774 women undergoing HPV typing test were collected as the source of cell specimens.Cervical tissue from the 298 subjects with HR-HPV positive and histopathological biopsy was the source of tissue specimens.HPV genotyping of cervical shed cells was detected by a combination of polymerase chain reaction(PCR)in vitro amplification and reverse point hybridization.SNP of HPV52 and HPV58 were analyzed by SnapGene and MEGA11.Hematoxylin-eosin(HE)staining was used to determine the grade of cervical lesions in the cervical tissue.Results HR-HPV among the tested women was mainly a single infection,with a positive rate of 20.36%,and the frequency distribution of HR-HPV infection was HPV52,HPV16,HPV58,HPV53,and HPV51.The HR-HPV dominance in cervical lesions was HPV16,HPV52,HPV58,HPV18,and HPV33.SNP and evolutionary tree analysis found that there were 7 SNP sites in type E6 and E7 of HPV52(Among them,A125T and A294G were the new mutation sites,and G350T and A379G had the highest mutation rates)and 3(of which T666C was the new mutation sites,and C751T and A801G had the highest mutation rate).Type HPV 52 was distributed in lineage B,with 3 and 8 SNP sites in regions E6 and E7 of HPV58,and the highest mutation site was T744G.The type HPV58 was distributed in lineage A.Conclusion The main positive types of the women in Anshun are HPV52,HPV16,and HPV58,and the main dominant types of cervical lesions are HPV16,HPV52,and HPV58.HPV52 and HPV58 have the specific SNP.