宫颈脱落细胞中人乳头瘤病毒58型感染及整合状况分析  

Human papillomavirus type 58 infection and integration in exfoliated cervical cells

作　　者：徐媛 张放 王静 杨佳宁 李淑英 XU Yuan;ZHANG Fang;WANG Jing;YANG Jianing;LI Shuying(School of Basic Medicine,North China University of Science and Technology,Hebei Key Laboratory for Chronic Diseases,Tangshan 063210,China)

机构地区：[1]华北理工大学基础医学院,河北省慢性疾病重点实验室,河北唐山063210

出　　处：《西安交通大学学报（医学版）》2022年第2期234-240,共7页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：河北省留学人员科技活动资助项目(No.CY201620)。

摘　　要：目的检测宫颈脱落细胞中人乳头瘤状病毒(human papillomavirus,HPV)感染状况,通过分析高危型HPV感染状况,筛查宫颈病变早期患者;并对HPV58型阳性标本的病毒载量及整合位点进行分析,探讨HPV58的病毒载量、整合状态与宫颈病变的关系,为宫颈病变的防治提供科学依据。方法收集宫颈脱落细胞标本及其临床病理资料,提取所收集标本基因组DNA及总RNA;应用HPV L1区通用引物进行多聚酶链反应(polymerase chain reaction,PCR)检测HPV总感染率,用HPV58 E6型特异性引物检测HPV58感染状况;荧光定量PCR检测HPV58 E6、E2及人管家基因β-actin的拷贝数,通过E2与E6比率确定HPV58整合状态;同时,应用HPV58癌基因转录扩增方法,分析整合型HPV58整合在宿主染色体的位置。结果1269例标本中检测到295例HPV阳性,阳性率23.2%,其中有34例标本HPV58阳性;HPV58阳性标本的整合状态分析显示,4例为纯游离型;8例为纯整合型;22例为整合与游离的混合型,宫颈病变程度与HPV58的病毒载量及整合状态无显著相关性;8例HPV58纯整合型标本,分别整合到患者1号、2号、5号、8号、17号染色体。结论所检宫颈脱落细胞标本中HPV总感染率为23.2%,随宫颈病变程度加重,HPV感染率增高;可通过高危型HPV感染筛查早期宫颈病变患者。Objective To detect the types of human papillomavirus(HPV)infection in exfoliated cervical cells to screen patients with early cervical lesions through HPV infection;analyze the viral load and integration sites of HPV58 positive specimens to explore the relationship of HPV58 viral load and integration status with cervical lesions in the hope of providing a scientific basis for the prevention and treatment of cervical lesions.Methods The exfoliated cervical cells and their clinicopathological data were collected,and genome DNA and total RNA were extracted from the collected specimen.HPV infections were detected using MY09/11 of HPVL1 region general primers and HPV58 was detected using HPV58 E6 specific primers.The copy numbers of HPV58 E6,E2 and human housekeeper geneβ-actin were detected by quantitative fluorescence PCR,and the integration status of HPV58 was determined by the ratio of E2 to E6.At the same time,the position of HPV58 integrated into the host chromosome was analyzed through oncogene transcription amplification.Results Totally 295 HPV positive cases were detected in 1269 exfoliated cervical cell samples,with the total infection rate of 23.2%.Among them 34 HPV58 positive were detected,which consisted of 4 samples of pure episome,8 samples of integration,and 22 cases of mixture(both integration and episome).Statistical analysis showed no significant correlation of the degree of cervical lesions with the viral load and integration status of HPV58.Eight HPV58 samples were integrated into chromosomes 1,2,5,8,and 17,respectively.Conclusion HPV positive rate was 23.2%in the exfoliated cervical cells,the HPV infection rate increased with the aggravation of cervical lesions.Patients with early cervical lesions can be screened through high-risk HPV infection.

关 键 词：宫颈病变 人乳头瘤状病毒 感染 整合 

分 类 号：R373[医药卫生—病原生物学]