Detection and typing of HPV by PCR and Restriction Fragment Length Polymorphisms (RFLPs) in tissue samples from human cervical lesions

Abstract

The identification and typing of HPVs (Human Papilloma Viruses) has become increasingly challenging, due to the numerous viral types that must be detected and their role in the process of cervical neoplasia. In the present study we used a combination of PCR amplification method and RFLP analysis in order to identify established or presumed novel HPV-types in various cervical lesions and consequently to evaluate its effectiveness in archival histopathological tissue samples. Forty two cases with histologic diagnosis of “pure” HPV-lesions [n=3], low grade intraepithelial lesions-LGSIL [n=16] and high grade intraepithelial lesions-HGSIL [n=23] were studied. Overall HPV-DNA was detected in 30 out of 42 cases analyzed [Detection rate: 71.4%]. High- risk HPV-types were detected in 15 of 23 HGSILs [65.1%], in 11 of 16 LGSILs [68.7%] and in one of 3 cases of “pure” HPV-lesions [33.3%]. Low risk HPV-types were detected in 1 of 23 HGSILs [4,3%] and in 1 of 16 LGSILs [6.2%]. One possibly novel HPV-type was detected in one case of LGSIL. HPV-typing by RLFP analysis was successful in 13 out of 30 positive cases [43.3%]. HPV-16 was identified in 6 cases [5 of HGSIL and 1 of LGSIL], HPV-31 in 3 cases [1 of HGSIL with microinvasion and 2 of LGSIL], HPV-33 in 1 case of HGSIL, HPV-56 in 1 case of LGSIL, HPV-61 in 1 case of HPVlesion and one possibly novel HPV-type in a case of LGSIL. This method could facilitate the sensitive identification of a broad spectrum of known and novel genital HPVs and may serve as an adjunct to existing traditional hybridization-based methods.