【作者】 蒋汉梁；

【导师】 陈智；

【作者基本信息】 浙江大学 ， 内科学， 2008， 博士

【摘要】 人乳头瘤病毒(Human Papillomavirus,HPV)感染是宫颈癌发生的必要因素,宫颈癌是严重威胁我国女性健康的妇科常见恶性肿瘤。流行病学研究表明,高危型HPV的持续感染是宫颈癌形成的最主要因素。早期发现宫颈高危型HPV感染与准确分型,及时进行早期干预治疗,是提高宫颈癌防治效果的有效途径。为了建立一种快速高通量的HPV分型检测方法,本研究以悬浮芯片技术为平台,分别设计HPV型特异性探针,并分别联结至不同编号的微球上,在不断优化杂交反应条件后,建立了一种快速高通量的HPV分型检测方法(HPVsuspension array,HPV-SA):通过对HPV基因型标准品的检测,证实本法所使用的探针具有高的特异性,各探针间未见有交叉反应;进一步对720例临床样本进行检测,证实HPV-SA能够快速准确地检测临床样本的HPV感染型别;以E7型特异性PCR法和DNA测序法作为检测的“金标准”,证实HPV-SA法的检测灵敏度为86.14%,特异度为98.26%,阳性预测值为95.08%,阴性预测值为92.92%,一致性为94.86%。为了获得本地区HPV亚型感染情况的信息,以及了解宫颈病变严重程度与HPV感染亚型之间的关系。本研究采用已建立的高通量HPV-SA法,对Normal,CINⅠ,CINⅡ,CINⅢ和Carcinoma等各种宫颈病理组织进行了HPV的感染情况研究;发现本地区HPV总体感染情况,是以HPV16的感染率为最高,其次为HPV58和HPV18;在Normal、CINⅠ、CINⅡ和CINⅢ组织以HPV16、58的感染为主,而在Carcinoma组织则以HPV16、18的感染为主;高危型HPV的单纯感染率要显著高于混合感染率;病变越是严重的组织,其高危型HPV的感染率就越高。这些信息的获得,为本地区更有针对性的宫颈防癌筛查策略的制定打下了基础,也为更具针对性的HPV疫苗的设计提供了重要的信息。更多还原

【Abstract】 Cervical cancer,a common gynecologic malignancy,is a serious threat to women’s health in china.Epidemiological studies have shown that the persistent infection of high-risk HPV is the most important factor for cervical cancer formation. The effective way to decrease the cervical cancer morbidity is to detect and classify the high-risk HPV infection in early time.In this study,we have established a rapid high-throughput HPV genotyping method by using the suspension array technology(HPV suspension array,HPV-SA). The specificity had been confirmed by the hybridization between probes and standard samples.Experiments show that HPV-SA can quickly and accurately detect 720 clinical samples.The detection accuracy of HPV using HPV-SA for 720 samples was as follows:sensitivity 86.14%,specificity 98.26%,positive predictive value 95.08%,the negative predictive value 92.92%,consistency 94.86%.In order to obtain subtypes information of HPV infection in the region,as well as understand the relationships between HPV subtypes and cervical lesions.We have studied on the HPV prevalence in different cervical lesions by using the HPV-SA method.Experiments show that the major prevalent high-risk HPV subtypes were HPV16,58 and 18 in normal,CINⅠ,CINⅡand CINⅢtissues,while in carcinoma tissue were HPV16 and 18.The single infection rate was significantly higher than multiple infections in all lesions.This information can improve the effect of cervical cancer screening,as well as provide important information for HPV vaccine design.更多还原