TYPING OF HUMAN PAPILLOMAVIRUSES BY CONSENSUS POLYMERASE CHAIN-REACTION AND A NONRADIOACTIVE REVERSE DOT-BLOT HYBRIDIZATION

A non-radioactive reverse dot blot hybridization method was developed for typing of human papillomavirus (HPV) consensus primer generated polymerase chain reaction (PCR) products in a single test. In the PCR biotin-14-dATP was incorporated into the amplified DNA, which was then used as a probe in hybridization with a membrane, on which different genital HPV types had been immobilized. Of cervical brush samples from women referred to a colposcopy clinic (n = 58) and from women attending a health control program (n = 14) which had been found positive by PCR with consensus HPV primers but negative using primers specific for the HPV types 6, 11, 16, 18, 31, 33 and 35, 25 (43%) and 3 (21%), respectively, could be typed by this method. The additional HPV types found were 34, 39, 40, 45, 52, 53, 56 and 58. Of the samples from the colposcopy clinic (n = 33) and the health control group (n = 11) which could not be typed, 23 and 5, respectively, showed HPV X which cross-hybridized with various HPV types under conditions of low stringency. It is possible to type by this fast and easy method consensus primer-generated PCR products of a wide range of HPV types or to verify the presence of HPV DNA of unknown types.