Detection and typing of minimal human papillomavirus DNA in plasma

ObjectiveFor clinical study of minute human papillomavirus (HPV) DNA such as those in plasma, a sensitive method that can detect a broad spectrum of HPV type is needed.MethodA nested polymerase chain reaction (PCR) method which utilized a consensus primer set nested to the widely utilized MY09/MY11 primer set was developed. The HPV genotype was determined by restriction digestion of the PCR product followed by agarose gel electrophoresis. DNA purified from cancer tissue, plasma and WBC of 17 patients of stage 1 or 2 squamous carcinoma of uterine cervix were examined.ResultsThis method readily detects a broad spectrum of at least ten genital types of HPV with a sensitivity of one viral copy. HPV DNA was detectable in all cervical cancer tissues and in 11 (65%) of the corresponding plasma, from which the genotype was successfully determined in 9 cases, all identical to that of primary cancer tissue.ConclusionThe high sensitivity and accuracy of this method has allowed detection of HPV in specimens of minimal viral load, such as in plasma in peripheral circulation of cervical cancer patients.