title: Degenerate and nested PCR: a highly sensitive and specific method for detection of human papillomavirus infection in cutaneous warts.
abstract: The role of human papillomavirus (HPV) in anogenital carcinogenesis is firmly established, but evidence that supports a similar role in skin remains speculative. Immunosuppressed renal transplant recipients have an increased incidence of viral warts and nonmelanoma skin cancer, and the presence of HPV DNA in these lesions, especially types associated with the condition epidermodysplasia verruciformis (EV), has led to suggestions that HPV may play a pathogenic role. However, differences in the specificities and sensitivities of techniques used to detect HPV in skin have led to wide discrepancies in the spectrum of HPV types reported. We describe a degenerate nested PCR technique with the capacity to detect a broad spectrum of cutaneous, mucosal, and EV HPV types. In a series of 51 warts from 23 renal transplant recipients, this method detected HPV DNA in all lesions, representing a significant improvement over many previously published studies. Cutaneous types were found in 84.3% of warts and EV types were found in 80.4% of warts, whereas 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 15 immunocompetent individuals. In summary, we have established a highly sensitive and comprehensive degenerate PCR methodology for detection and genotyping of HPV from the skin and have demonstrated a diverse spectrum of multiple HPV types in cutaneous warts from transplant recipients. Studies designed to assess the significance of these findings to cutaneous carcinogenesis are under way.
authors.affiliation: Department of Academic Dermatology, Royal Hospitals NHS Trust, London, United Kingdom. caharwood@doctors.org.uk
authors.forename: C A
authors.initials: CA
authors.lastname: Harwood
authors.forename: P J
authors.initials: PJ
authors.lastname: Spink
authors.forename: T
authors.initials: T
authors.lastname: Surentheran
authors.forename: I M
authors.initials: IM
authors.lastname: Leigh
authors.forename: E M
authors.initials: EM
authors.lastname: de Villiers
authors.forename: J M
authors.initials: JM
authors.lastname: McGregor
authors.forename: C M
authors.initials: CM
authors.lastname: Proby
authors.forename: J
authors.initials: J
authors.lastname: Breuer
mesh_terms: D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D005069:Evaluation Studies as Topic; D005260:Female; D006801:Humans; D007121:Immunocompetence; D008297:Male; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D014181:Transplantation Immunology; D014412:Tumor Virus Infections / Q000175:diagnosis / Q000821:virology*; D014860:Warts / Q000821:virology*
chemical_list: D017931:DNA Primers