CAJ | 학술논문

目的研究广州东部妇女中人乳头瘤病毒16型(HPV-16)宫颈感染分布,分析其早基因E6/E7的多态性,分析L1和E6基因定量与病程的关系.方法通过导流杂交基因芯片技术检测宫颈脱落细胞的HPV-16感染;通过特异性扩增获取病毒早基因E6/E7序列,克隆测序并进行多态性分析;荧光定量PCR技术对E6基因和L1基因进行定量分析.结果 806例宫颈脱落细胞样本中HPV-16感染阳性36例(4.5%),其中18例(50.0%)宫颈细胞发生高度以上病变;7例(4例低度或以下病变,3例高度以上病变或浸润癌)阳性标本得到E6/g7序列有15个位点分别出现变异;高度病变组(A组,11例)与低度或以下病变组(B组,14例)的L1基因和E6基因定量数据对数值均有显著差异(P<0.05),但L1/E6比值差异无统计学意义(P=0.19).结论本地区在17～62岁妇女中HPV-16感染阳性发生率约4.5%,50.0%发生高度以上宫颈病变,本研究显示病毒基因拷贝数与宫颈病变程度可能有关,L1/E6比值未能提示病毒整合的发生.
목적 연구엄주동부부녀중인유두류병독16형(HPV-16)궁경감염분포,분석기조기인E6/E7적다태성,분석L1화E6기인정량여병정적관계.방법 통과도류잡교기인심편기술검측궁경탈락세포적HPV-16감염;통과특이성확증획취병독조기인E6/E7서렬,극륭측서병진행다태성분석;형광정량PCR기술대E6기인화L1기인진행정량분석.결과 806례궁경탈락세포양본중HPV-16감염양성36례(4.5%),기중18례(50.0%)궁경세포발생고도이상병변;7례(4례저도혹이하병변,3례고도이상병변혹침윤암)양성표본득도E6/g7서렬유15개위점분별출현변이;고도병변조(A조,11례)여저도혹이하병변조(B조,14례)적L1기인화E6기인정량수거대수치균유현저차이(P<0.05),단L1/E6비치차이무통계학의의(P=0.19).결론 본지구재17～62세부녀중HPV-16감염양성발생솔약4.5%,50.0%발생고도이상궁경병변,본연구현시병독기인고패수여궁경병변정도가능유관,L1/E6비치미능제시병독정합적발생.
Objective To investigate the distribution of human papillomavirus (HPV) type 16 in women cervical infection in eastern Guangzhou, polymorphism of E6/E7 gene and association of gene dosage with disease progression. Methods Flow-through hybridization and gene chips were applied in HPV sub-type identification to screen out HPV-16 positive samples from cervical epithelium samples. HPV-16 E6/E7 gene was amplified through PCR with specific primers. The PCR products were cloned into pMD18-T vectors and fragments were determined through sequencing. Polymorphism analysis were performed through align-ment tools. Fluorescence quantitive PCR were used for the detection of viral E6 gene and L1 gene. Results Thirty-six (4.5%) HPV-16 positive samples were screened out through flow-through hybridization from 806 cervical epithelium samples. HSIL and above happened in 18 (50.0%) of the 36 HPV-16 positive patients. Within E6/E7 gene sequences from 7 selected samples, we found 15 sites with variances and 8 of them would cause coding amino acid change. HIL group (A, 11 cases) and LSIL group (B, 14 cases) possess significantly different gene dosage of both viral E6 gene and LI gene (P <0.05). The ratios of L1/E6 be-tween the 2 groups was not significantly different(P=0.19). Conclusion HPV-16 cervical infection oc-curs in 4.5% women (17-62 years old) in eastern Guangzhou. HIL or above accompany with half of the HPV 16 infected women. Viral load is probably associated with cervical HSIL, though L1/E6 ratios do not suggest viral integration.