CHARACTERIZATION OF A PUTATIVE NEW HPV GENOMIC SEQUENCE FROM A CERVICAL LESION USING L1 CONSENSUS PRIMERS AND RESTRICTION-FRAGMENT-LENGTH-POLYMORPHISM

Various methods have been proposed for HPV detection and typing. Preva lence and distribution among types have varied depending upon the meth ods used and the populations studied. We have applied the polymerase c hain reaction (PCR) followed by a Restriction Fragment Length Polymorp hism (RFLP) analysis using the MY09/MY11 primers for detection of HPV in cervicovaginal lavages obtained from 323 patients who were referred to our Clinical Department either for genital complaints or an abnorm al PAP smear. We assessed (i) the prevalence of HPV and (ii) the relia bility of RFLP-typing. For the latter, 35 PCR-HPV products were sequen ced. HPV-DNA was detected in 40/197 (20.3%) patients with normal cytol ogy, 86/111 (77.5%) with LSIL and 11/15 (73.3%) with HSIL. HPV-16 was the most common type detected in normal cervical cytology samples (10/ 40, 25%), whereas HPV 16 and 18 were detected in 36/97 (37.1%) of the LSIL and HSIL patients, evidencing the presence of these high-risk HPV types not only in malignant conditions. Results obtained after partia l nucleotide sequencing confirmed the results obtained by RFLP analysi s. In this study, a putative new HPV fragment (GA6053) was identified. Its closest homology to other known HPV types is 73.8% to HPV-62, 73. 0% to HPV-61 and 67.7% to HPV-18. The use of degenerate primers, in co njunction with RFLP, proved to be a reliable method for HPV detection and typing. (C) 1997 Elsevier Science B.V.