PCR-BASED NATURAL HISTORY STUDY OF HPV INFECTIONS

基于 PCR 的 HPV 感染自然史研究

• 批准号: 3148001
• 负责人: Cosette M Wheeler
• 金额: $ 13.77万
• 依托单位: UNIVERSITY OF NEW MEXICO
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-30 至 1995-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3148001
• 关键词: Hispanic Americans; Native Americans; biomarker; caucasian American; cervical /vaginal smear; cervix neoplasms; clinical chemistry; computer assisted sequence analysis; cytology; diagnosis design /evaluation; disease /disorder proneness /risk; endonuclease; female reproductive system disorder diagnosis; human papillomavirus; human subject; molecular cloning; neoplasm /cancer diagnosis; neoplasm /cancer invasiveness; nucleic acid probes; nucleic acid sequence; polymerase chain reaction; preneoplastic state; questionnaires; southern blotting; virus classification; virus related neoplasm /cancer

To date, 60 different types of human papillomaviruses (HPVs) have been isolated and characterized with the aid of molecular cloning systems. Over twenty of these viral isolates have been associated with benign and malignant lesions of the anogenital tract. The presence of HPV in premalignant and invasive cancers, particularly of the cervix, is believed to be reflective of the oncogenic potential of these viruses. We propose to conduct a natural history study of HPV infections in high- risk minority populations including Native American, Hispanic, and Non-Hispanic white women. As part of these investigations we will establish a PCR-based HPV L1/E6 detection system that correlates with established Southern blot criteria of HPV type. This system will be utilized to determine the prevalence of various HPV types and their potential relationship to cervical disease in these randomly screened populations. Additionally, we will evaluate cervical sampling methodologies and analyze biological and behavioral factors affecting the detection of HPVs in serial cervical specimens. No diagnostic method currently available provides a sensitive, specific and reproducible approach to identifying and characterizing this highly heterogeneic group of viruses. A diagnostic possessing these characteristics which has been developed systematically in defined populations is needed before it can be determined if specific HPVs act as etiologic agents, cofactors or markers of developing cervical neoplasias. Prevalent novel HPVs identified in the populations studied will be subjected to direct automated DNA sequence analysis and the resultant data utilized to develop improved oligonucleotide probe systems. HPV types and specific DNA sequences will be evaluated as potential risk factors or markers of cervical disease. A simplified strategy for identification of known and novel HPV isolates which utilizes restricition map analysis of HPV L1 and E6/E7 PCR products will be developed. HPV E6-E7 PCR products obtained from "normal", preinvasive, and invasive cervical lesions will be directly cloned and evaluated for in vitro transforming activities. Pathological findings and in vitro transforming activities will be compared between and within multiple HPV types in an attempt to identify HPV-specific sequences associated with cervical neoplasia.

迄今为止，已发现 60 种不同类型的人乳头瘤病毒 (HPV) 借助分子克隆系统进行分离和表征。 这些病毒分离株中有二十多种与良性和 肛门生殖道的恶性病变。 HPV 的存在 癌前和浸润性癌症，特别是子宫颈癌， 据信反映了这些病毒的致癌潜力。 我们建议对高危人群中的 HPV 感染进行自然史研究。 少数族裔面临风险，包括美洲原住民、西班牙裔和 非西班牙裔白人女性。 作为这些调查的一部分，我们将 建立基于 PCR 的 HPV L1/E6 检测系统，该系统与 建立了 HPV 类型的 Southern 印迹标准。 该系统将 用于确定各种 HPV 类型的患病率及其 在这些随机筛查中与宫颈疾病的潜在关系 人口。 此外，我们将评估宫颈取样 方法论并分析影响的生物和行为因素 连续宫颈标本中 HPV 的检测。 无诊断 目前可用的方法提供了一种敏感的、特异的和 可重复的方法来识别和表征这种高度 异质病毒群。 拥有这些的诊断 已在定义中系统地开发的特征 在确定特定 HPV 是否起作用之前，需要对人群进行检测 作为宫颈癌发展的病因、辅助因子或标志物 肿瘤。 在研究人群中发现了流行的新型 HPV 将进行直接自动化 DNA 序列分析 所得数据用于开发改进的寡核苷酸探针 系统。 HPV 类型和特定 DNA 序列将被评估为 宫颈疾病的潜在危险因素或标志物。 一个简化的 识别已知和新型 HPV 分离株的策略 利用 HPV L1 和 E6/E7 PCR 产物的限制性图谱分析将 得到开发。 从“正常”获得的HPV E6-E7 PCR产物， 浸润前和浸润性宫颈病变将被直接克隆并 评估体外转化活性。 病理结果 和体外转化活性将在之间和内部进行比较 多种 HPV 类型，试图鉴定 HPV 特异性序列 与宫颈肿瘤有关。