Degenerate and nested PCR: a highly sensitive and specific method for detection of human papillomavirus infection in cutaneous warts

The role of human papillomavirus (HPV) in anogenital carcinogenesis is firm ly established, but evidence that supports a similar role in skin remains s peculative, Immunosuppressed renal transplant recipients have an increased incidence of viral warts and nonmelanoma skin cancer, and the presence of H PV DNA in these lesions, especially types associated with the condition epi dermodysplasia verruciformis (EV), has led to suggestions that HPV may play a pathogenic role. However, differences in the specificities and sensitivi ties of techniques used to detect HPV in skin have led to wide discrepancie s in the spectrum of BPV types reported. We describe a degenerate nested PC R technique with the capacity to detect a broad spectrum of cutaneous, muco sal, and EV HPV types. In a series of 51 warts from 23 renal transplant rec ipients, this method detected HPV DNA in all lesions, representing a signif icant improvement over many previously published studies. Cutaneous types w ere found in 84.3% of warts and EW types were found in 80.4% of warts, wher eas mucosal types were detected in 27.4% of warts, In addition, the method allowed codetection of two or more distinct HPV types in 94.1% of lesions, In contrast, single HPV types were detected in all but 1 of 20 warts from 1 5 immunocompetent individuals. In summary, we have established a highly sen sitive and comprehensive degenerate PCR methodology for detection and genot yping of HPV from the skin and have demonstrated a diverse spectrum of mult iple HPV types in cutaneous warts from transplant recipients, Studies desig ned to assess the significance of these findings to cutaneous carcinogenesi s are under way.