Detection and identification of human papillomavirus DNA in genital tract by digoxigenin labelling using the...

We propose here a simple and rapid method of Human Papillomavirus (HPV) detection applied to genital samples. The method consists of a polymerase chain reaction (PCR) of samples using a consensus primer ser (Snijders et al., 1990) with labelling by Dig-dUTP (Digoxigenin deoxyuridine triphosphate) during amplification. Type identification was carried out by hybridization of the labelling PCR product with a panel of different plasmidic HPVs dotted beforehand on nylon membranes. Forty-five genital samples were tested with this method (30 samples with evident signs of HPV infection, and 15 cervical scrapes which had a normal colposcopic examination and no cytologic signs of HPV infection). We found good sensitivity and specificity levels. The labelling method during PCR did not modify the sensitivity of PCR. Forty-five genital samples were analyzed by this technique and compared with the results of dot-blot and two PCR methods using different primers. They showed a good correlation except in four samples (high grade cervical lesions) which seemed to show a partial deletion or rearrangement. Our method, used for the first time in HPV detection, needed just one PCR and one dot-blot, thus saving an important amount of time and also decreasing examination costs.