DETECTION OF HUMAN PAPILLOMAVIRUS (HPV) TYPE-47 DNA IN MALIGNANT LESIONS FROM EPIDERMODYSPLASIA-VERRUCIFORMIS BY PROTOCOLS FOR PRECISE TYPING OF RELATED HPV DNAS

Our discovery of human papillomavirus type 47 (HPV47) in benign lesion s from a patient suffering from epidermodysplasia verruciformis prompt ed us to examine whether the viral DNA also resided in malignant lesio ns from the same patient. By using newly devised protocols for amplify ing a group of epidermodysplasia verruciformis-associated HPV DNAs by PCR and differentially identifying them by reverse-phase dot blot hybr idization, we demonstrated that HPV47 DNA, but not other HPV DNAs of t he group, was abundant (about 10(3) copies per diploid amount of cell DNA) in DNAs prepared from three carcinomas. Using DNA from one or the se carcinomas, we also confirmed that DNA of HPV5, HPV14 or HPV21, det ected in significant amounts in DNAs from benign lesions from the pati ent, were present only in negligible amounts or not at all, The result s suggest the involvement of HPV47 DNA in tumorigenesis, Furthermore, we demonstrated by the Southern technique that most, if not all, of th e HPV47 DNA consists of either a unit (or a nongrossly deleted unit) l ength of the viral genome carrying no (or no gross) internal rearrange ments or tandem repeats, This and other results obtained by this techn ique indicated that a considerable amount of the viral DNA resides as a circular monomer a unit length of the viral genome in carcinoma cell s, while the remainder reside as catenanes, concatemers, or both, The concatemers were considered more likely to be replicated without integ ration into cellular DNA than to be integrated, because no bands for t he corresponding fragments including integration sites were detected b y treatment with restriction enzymes that would have produced such fra gments.