ASSAYS TO DETECT AND TYPE HPV DNA IN ORAL CANCER

点击翻译按钮获取中文摘要

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.