DEVELOPMENT OF TYPE SPECIFIC HUMAN PAPILLOMAVIRUS PROBES

从世界各地的一些研究中可以清楚地看出， 女性生殖道的乳头瘤病毒（HPV）感染是一种重要的 宫颈癌的早期症状有哪些？ 因为HPV可以 存在于正常组织中，无病理学表现，可能是潜伏的病毒 感染，我们建议测试构建核酸的可行性 探针，这将允许开发一个简单的杂交测试， HPV的检测。 在第一阶段的提议中，我们计划构建特定的探针， 检测不同的HPV类型，但彼此没有同源性。 的 由于不同HPV之间的同源性，需要型特异性探针 类型，这使得全长基因组探针对于测试是不期望的 人体样本 为了开发这些探针，我们将使用可用的序列数据 异源双链体作图的结果表明， 不同HPV类型之间的区域。 调查领域将是1） 亚基因组型特异性RNA和DNA探针的开发2）复合物 探针，由串联的类型特异性序列组成，和3）合成的 寡核苷酸探针。 这些探头的性能将通过以下方式进行评价： 克隆病毒DNA和临床病毒DNA的Southern杂交和斑点杂交分析 样品

It is clear from a number of studies throughout the world that Human Papillomavirus (HPV) infection of the female genital tract is a significant risk factor in the development of cervical cancer. Since HPV can be present in normal tissue showing no pathology, perhaps as a latent viral infection, we propose to test the feasibility of constructing nucleic acid probes that will permit the development of a simple hybridization test for the detection of HPVs. In this phase I proposal we plan to construct specific probes that can detect the different HPV types but have no homology to each other. The necessity for type specific probes is due to homology between different HPV types, which renders full length genomic probes undesirable for testing human samples. To develop these probes we will use available sequence data and results from heteroduplex mapping which indicate the nonhomologous regions between different HPV types. Areas of investigation will be 1) development of subgenomic type-specific RNA and DNA probes 2) composite probes, consisting of type-specific sequences in tandem and 3) synthetic oligonucleotide probes. Performance of these probes will be evaluated by Southern and Dot blot analysis on both cloned viral DNA and clinical samples.