ASSAYS TO DETECT AND TYPE HPV DNA IN ORAL CANCER

人乳头瘤病毒(HPV)感染与肺癌发生的关系 从流行病学和医学角度来看，宫颈已经牢固确立 在实验室里。几种HPV亚型现在被认为是 宫颈癌的致病因素。然而，HPV的相关性 口腔癌还没有得到充分的调查。由Southern blotting 技术，已在口腔粘膜中检测到HPV-32、HPV-13和HPV-6 局灶性上皮增生症。近年来，人乳头瘤病毒在临床上的检测 宫颈样本是通过使用聚合酶链来完成的 反应(PCR)扩增技术。 在此阶段I提案中，我们描述了一种创新的筛选策略 口腔癌标本中是否存在HPV DNA。通过使用普通的聚合酶链反应 与HPV基因高度保守的L1区杂交的引物， 所有HPV亚型均可在一次聚合酶链式反应中扩增。样本： 被这种“快速筛查”试验判断为阳性的人将使用 不同的聚合酶链式反应L1共有引物，通过杂交复制 带有不同HPV亚型DNA探针的过滤器，亚型将是 下定决心。通过这项技术，HPV序列的存在和 可以确定临床口腔癌标本中HPV的特异性亚型。

Association of human papillomaviruses (HPV) infection an carcinogenesis of the uterine cervix has been firmly established both epidemiologically and in the laboratory. Several HPV subtypes are now considered to be contributing factors for cervical cancer. However, the association of HPV and oral cancer has not been fully investigated. By the Southern blotting technique, HPV-32, HPV-13 and HPV-6 have been detected in oral mucosa in focal epithelial hyperplasia. Recently, detection of HPV in clinical cervical samples was accomplished by the use of the polymerase chain reaction (PCR) amplification technique. In this Phase I proposal, we describe an innovative strategy for screening oral cancer samples for the presence of HPV DNA. By using general PCR primers which hybridize to the highly conserved L1 region of the HPV gene, all HPV subtypes can be amplified in a single PCR reaction. Samples which are judged positive by this "quick screen" assay will be amplified using a different set of PCR L1 consensus primers and, by hybridizing replicate filters with DNA probes of various HPV subtypes, the subtype will be determined. By this technique, both the presence of HPV sequences and the specific HPV subtype in clinical oral cancer specimens can be determined.