DETECTION OF HUMAN PAPILLOMAVIRUS DNA IN CERVICAL LAVAGE SPECIMENS BYA NONISOTOPIC CONSENSUS PCR ASSAY

A gene amplification method that combines PCR with an enzyme immunoass ay (PCR-EIA) for quantitation of amplified DNA was developed for the d etection of human papillomavirus (HPV), Samples were amplified with co nsensus primers MY09 and MY11, Amplified DNA products were reacted in solution with type-specific nested RNA probes labelled with digoxigeni n-11-UTP. Hybrids were captured on a microtiter plate coated with an a ntidigoxigenin antibody, Bound DNA-RNA hybrids were quantitated by the addition of an alkaline phosphatase-labelled monoclonal antibody dire cted against DNA-RNA hybrids and a fluorogenic substrate, The detectio n limit of PCR-EIA was six copies of HPV type 18 DNA in the original s pecimen, The assay was used to assess HPV infection of the uterine cer vixes of 65 women referred to a colposcopy clinic, In 66 cervicovagina l lavage specimens, all 23 HPV strains detected by a standard isotopic PCR assay were also detected by the PCR-EIA (sensitivity, 100%; 95% c onfidence interval, 85.2 to 100%), Forty-two of the 43 samples that di d not contain HPV types 6/11, 16, 18, 31, 33, 35, and 45 were also neg ative by PCR-EIA, for a specificity of 97.7%. Low-level cross-reactivi ty was encountered between HPV types 18 and 45 as well as between type s 33 and 58, PCR-EIA provides a convenient means of objectively measur ing PCR-amplified HPV DNA from common genital HPV types.