GROUP-SPECIFIC DIFFERENTIATION BETWEEN HIGH- AND LOW-RISK HUMAN PAPILLOMAVIRUS GENOTYPES BY GENERAL PRIMER-MEDIATED PCR AND 2 COCKTAILS OF OLIGONUCLEOTIDE PROBES

In recent years, general primer-mediated PCR assays have been develope d to detect a broad spectrum of human papillomavirus (HPV) genotypes. In this study, a procedure enabling a simple group-specific differenti ation of high-risk (HPV-16, -18, -31, -33, -35, -39, -45, -51, -52, -5 4, -56, and -58) and low-risk (HPV-6, -11, -34, 40, -42, -43, and 44) HPVs following an HPV general primer-mediated (GP5+/GP6+) PCR is prese nted. By computer-assisted sequence analysis, oligonucleotides (30-mer s) specific for 19 different HPV genotypes were selected from the inte rnal part of the 150-bp GP5+/GP6+-amplified region. These oligo probes were tested for specificity in a Southern blot analysis of PCR produc ts derived from the same panel of HPV types. No cross-hybridizations w ere found. The sensitivities of the oligo probes varied from the femto gram level for the well-amplified HPV types like HPV-16 and -18 to the picogram level for the less-well-amplified HPV types like HPV-39 and -51. These sensitivities were reached when the oligo probes were appli ed both individually and in a cocktail. On the basis of these results, two cocktail oligo probes that enabled a specific and sensitive diffe rentiation between low- and high-risk HPV types were composed.