DEVELOPMENT OF TYPE SPECIFIC HUMAN PAPILLOMAVIRUS PROBES

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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.