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，已经有60种不同类型的人乳头瘤病毒（HPV） 借助分子克隆系统进行了分离和表征。 这些病毒分离株中有20多种与良性和 肛门生殖道的恶性病变。 HPV的存在 前癌和侵入性癌症，特别是子宫颈的癌症是 被认为反映了这些病毒的致癌潜力。 我们建议在高位中对HPV感染进行自然史研究 风险包括美国原住民，西班牙裔和 非西班牙裔白人妇女。 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 建立的HPV类型的Southern印迹标准。 这个系统将是 用于确定各种HPV类型及其的患病率 这些随机筛查中与宫颈疾病的潜在关系 人群。 此外，我们将评估宫颈采样 方法论并分析影响的生物学和行为因素 串行宫颈标本中HPV的检测。 没有诊断 当前可用的方法提供了一种敏感，具体和 可重现的方法来识别和表征这一高度 异质性病毒组。 拥有这些的诊断 已定义的系统开发的特征 如果特定的HPV ACT，则需要在确定它之前需要种群 作为病因学剂，辅助因子或宫颈的标志物 肿瘤。 在研究的人群中确定的普遍新颖的HPV 将进行直接自动DNA序列分析和 最终的数据用于开发改进的寡核苷酸探针 系统。 HPV类型和特定的DNA序列将被评估为 宫颈疾病的潜在危险因素或标志物。 简化 鉴定已知和新型HPV分离株的策略 利用HPV L1和E6/E7 PCR产品的修复图分析将 被开发。 从“正常”获得的HPV E6-E7 PCR产品， 侵入性和侵袭性宫颈病变将直接克隆，并且 评估体外转化活动。 病理发现 并将在体外转化活动之间进行比较 尝试识别HPV特异性序列的多种HPV类型 与宫颈肿瘤有关。