Ultrasensitive detection of HPV DNA in clinical specimens by SPF-PCR and identification of HPV genotypes by reverse hybridization

Detection of HPV-DNA is widely used as a diagnostic method. However, the efficacy and accuracy of the molecular tools directly determines the clinical relevance of HPV testing and its role in patient management. We developed a novel general primer set (designated SPF) for broad spectrum detection of HPV, as well as a reverse hybridization method (LiPA) to identify 25 anogenital HPV genotypes. A large number of clinical samples, including cervical scrapes and (paraffin-embedded) biopsy specimens were tested and showed that the SPF-LiPA system (1) is ultrasensitive, (2) allows detection of a broad spectrum of HPV genotypes in various clinical materials (3) permits sensitive and specific identification of 25 genotypes. The method is particularly effective for simultaneous detection of multiple genotypes. In conclusion, this method may provide an effective tool for further analysis of the clinical relevance, epidemiology, and treatment of HPV infections.