Assays to Detect and Type HPV DNA in Oral Cancer

Association of human papillomaviruses (HPV) infection an carcinogenesis of the uterine cervixhas been firmly established both epidemiologically and in the laboratory. Several HPV subtypes are nowconsidered to be contributing factors for cervical cancer. However, the association of HPV and oralcancer has not been fully investigated. The Southern blotting technique, HPV-32, HPV-13 and HPV-6have been detected in oral mucosa in focal epithelial hyperplasia. Recently, detection of HPV in clinicalcervical samples was accomplished by the use of the polymerase chain reaction (PCR) amplificationtechnique. We describe an innovative strategy for screening oral cancer samples for the presence ofHPV DNA. By using general PCR primers which hybridize to the highly conserved L1 region of the HPVgene, all HPV subtypes can be amplified in a single PCR reaction. Samples which are judged positiveby 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 oralcancer specimens can be determined.