ANALYSIS OF PROSTATE TISSUE DNA FOR THE PRESENCE OF HUMAN PAPILLOMAVIRUS BY POLYMERASE CHAIN-REACTION, CLONING, AND AUTOMATED SEQUENCING

We have analysed the DNA from 24 prostate tissue biopsies, spanning a range of Gleason grading from benign to grade 5 and mixed randomly wit h cervical cancer samples of known human papillomavirus (HPV) status, for the prevalence of HPV DNA, in a double-blind study to ensure compl ete objectivity. Polymerase chain reactions (PCR) were performed using general E1 open reading frame primers for HPV under low stringency co nditions, in addition to reactions containing primers specific for HPV 16, E2, and E6 open reading frames under higher, more stringent PCR co nditions. The presence of cellular DNA was verified by the use of prim ers for hypoxanthine guanine phosphoribosyl transferase. DNA bands wer e not detected in the prostate biopsies using the HPV16-specific prime rs under high-stringency PCR conditions, however a predominant band in the 400 bp region was observed in 15 of the prostate biopsies using t he general primers and the low annealing temperature of 40 degrees C. This fragment was excised and cloned into the pT7 blue vector and the sequence of the insert determined. Although the cloned sequences initi ated and terminated with the two authentic PCR primers, they did not c ontain a significant HPV-related open reading frame. Our results indic ate that HPV type 16 and closely related types, as detected by the gen eral primer pair, are unlikely initiators of prostate carcinogenesis w ithin our population. (C) 1997 Wiley-Liss, Inc.