DETECTION OF HUMAN PAPILLOMAVIRUS IN CERVICAL INTRAEPITHELIAL NEOPLASIA, USING IN-SITU HYBRIDIZATION AND VARIOUS POLYMERASE CHAIN-REACTION TECHNIQUES

One hundred and forty-eight randomly chosen neutral-buffered formaldeh yde-fixed cervical biopsies in which cervical intra-epithelial neoplas ia (GIN) I-III had been diagnosed were tested for HPV (human papilloma virus) DNA by in situ hybridization (ISH) and polymer ase chain react ion (PCR). For ISH, we utilized a biotinylated panprobe and type-speci fic, genomic probe sets. For PCR, we used the general primers GP5/GP6 and their recently described, elongated version GP5+/GP6+, and include d the modification of hot-start PCR. Amplified DNA was detected by gel electrophoresis and slot blot hybridization. The positivity rate of I SH was 59% for all biopsies and 69%, 62% and 46% for CIN I, II and III , respectively. The sensitivity of GP5/GP6 was 74% with cold-start PCR and 78% with hot-start PCR. When GP5+/GP6+ was used, the sensitivity increased to 89% with cold-start PCR and to 95% with hot-start PCR. Ba sed on the most sensitive PCR technique, HPV detection was 93%, 95% an d 96% in CIN I, II and III, respectively. The number of HPV types decr eased with the severity of the lesion, and HPV 16 was the predominant type. Multiple HPVs were rare and almost all HPV-positive cases could be typed. ISH and slot blot hybridization correlated well regarding HP V typing specificity. Our results confirm that distinct HPV types are present in a high proportion of cases of GIN. The sensitivity of ISH i s lower than that of PCR. Furthermore, the modified general primers GP 5+/GP6+ give a higher yield than GP5/GP6, while hot-start PCR increase s sensitivity even further.