ASSAYS TO DETECT AND TYPE HPV DNA IN ORAL CANCER

口腔癌中 HPV DNA 的检测和分型分析

• 批准号: 3493627
• 负责人: TERESA A LEHMAN
• 金额: $ 7.23万
• 依托单位: BIOSERVE BIOTECHNOLOGIES, LTD
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-02-01 至 1994-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3493627
• 关键词: diagnosis design /evaluation; globin; human genetic material tag; human papillomavirus; human tissue; method development; neoplasm /cancer diagnosis; nucleic acid probes; nucleic acid sequence; oligonucleotides; oral pharyngeal disorder diagnosis; oral pharyngeal neoplasm; polymerase chain reaction; tissue /cell culture; virus DNA; virus diseases; virus genetics; virus protein

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.

人乳头瘤病毒感染与乳腺癌发生的关系 子宫颈在流行病学上和 在实验室 目前认为几种HPV亚型 宫颈癌的病因有哪些 然而，HPV的相关性 口腔癌还没有得到充分的研究。通过Southern印迹法 HPV-32、HPV-13和HPV-6在口腔粘膜中的检出率分别为 局灶性上皮增生 近年来，HPV检测在临床 使用聚合酶链完成宫颈样本 反应（PCR）扩增技术。 在这第一阶段的建议，我们描述了一个创新的战略， 口腔癌样本中HPV DNA的存在。 通过使用通用PCR 与HPV基因的高度保守的L1区杂交的引物， 所有HPV亚型都可以在单一PCR反应中扩增。 的样品 通过这种“快速筛选”试验判断为阳性的，将使用 不同组的PCR L1共有引物，并通过杂交重复 过滤器与各种HPV亚型的DNA探针，亚型将是 测定 通过这种技术，HPV序列的存在和 可以确定临床口腔癌标本中的特定HPV亚型。