DETECTION AND TYPING OF HUMAN PAPILLOMAVIRUSES IN MUCOSAL AND CUTANEOUS BIOPSIES FROM IMMUNOSUPPRESSED AND IMMUNOCOMPETENT PATIENTS AND PATIENTS WITH EPIDERMODYSPLASIA-VERRUCIFORMIS - A UNIFIED DIAGNOSTIC-APPROACH

Aim-To develop a unified diagnostic approach for the detection of huma n papillomavirus (HPV) DNA in skin and mucosal biopsies from both immu nocompetent and immunosuppressed individuals using a degenerate polyme rase chain reaction (PCR) method. Methods-The sensitivity and specific ity of three published degenerate primer sets (HVP2/B5 and F14/B15; MY 09/MY11; CP62/69 outer and CP65/68 nested primer pairs) were evaluated in PCR reactions with serial dilutions of 12 representative cloned HP V types. This combination of primers was then used to detect HPV DNA i n 49 benign and malignant lesions of cutaneous and mucosal origin from immunosuppressed, immunocompetent, and epidermodysplasia verruciformi s (EV) patients, and compared with detection rates using single primer sets alone. Results-The observed sensitivity of MY09/MY11 and CP62/69 +CP65/68 was high for mucosal and EV HPV types, respectively. The sens itivity of all primer sets for cutaneous types was low but nonetheless the use of this combination of primers allowed HPV DNA detection in a ll of the benign warts analysed. Several mixed infections were also id entified. A high prevalence of HPV DNA was similarly detected in squam ous cell carcinomas from immunocompromised patients; the HPV types fou nd were exclusively EV related. Conclusions-The use of a combined dege nerate primer PCR approach considerably improves HPV DNA detection ove r individual primer sets and allows detection of mixed infections. The findings may help explain the discrepancies in published reports rela ting to HPV DNA detection in benign and malignant skin lesions. Furthe r modifications to this method are in progress which should significan tly improve comprehensive HPV detection and typing for diagnostic purp oses.