Detection of human papillomaviruses in cervical neoplasias using multiple sets of generic polymerase chain reaction primers

Objective. The aim of this study was to evaluate precisely the differences in the spectra of human papillomavirus (HPV) types detected by different ge neric primer pairs commonly used for detection of this extraordinarily hete rogeneous virus. Methods. Three sets of polymerase chain reaction (PCR) primers for the L1 o pen reading frame (ORF) and two sets for E6/E7 ORFs were used to detect HPV s in DNAs from 107 cervical tissues, including 77 cervical neoplasias. HPV types were determined by analysis of restriction fragment length polymorphi sms (RFLPs) and nucleotide sequencing. Results. A high overall detection rate of HPV in cervical neoplasias (76/77 , 98.7%) was achieved by polymerase chain reaction (PCR) amplification with multiple sets of generic primers, while the detection rate for each indivi dual primer pair varied from 48/77 (62%) to 70/77 (91%). Only in 34 of 77 c ases (44%) were HPV DNAs positive for all sets of primer pairs. Further det ermination of HPV types by RFLPs and nucleotide sequencing showed inconsist encies between the PCR primer pairs used. Conclusion. Our study revealed that the HPV detection rate is critically af fected by the choice of PCR primers, and that appropriate use of combinatio ns of generic PCR primer sets followed by RFLP analyses is both necessary a nd sufficient for typing most HPVs in cervical lesions. More precise method s such as sequencing would be necessary in only a few cases. (C) 2001 Acade mic Press.