Improved detection of cutaneous human papillomavirus DNA by single tube nested 'hanging droplet' PCR.
A single tube nested 'hanging droplet' PCR was developed for detection of cutaneous human papillomavirus (HPV) DNA of the phylogenetic group B1. The nested PCR was compared with a single round PCR method by testing 56 fresh biopsies from Australian skin tumour patients. HPV DNA was detected in 64% (36/56) of the biopsies by nested PCR and in 30% (17/56) by single round PCR (P<0.001). HPV DNA was more often detected by nested PCR than by single round PCR in basal cell carcinoma [62% (16/26) vs. 19%; (5/26); P=0.003], squamous cell carcinoma [43% (7/16) vs. 25% (4/16)] and in solar keratosis [93% (13/14) vs. 57% (8/14); P=0.038]. The nested PCR and the single round PCR system detected 26 and 11 different HPV types/putative types/subtypes, respectively. Multiple types were found in eight samples by the nested PCR and two samples by single round PCR. The nested HPV PCR is more sensitive and capable of amplifying a broad spectrum of HPV types from skin tumours, but further improvements are needed before all HPV infections in skin can be detected by a single assay.
Forslund
Ola
O
Department of Medical Microbiology, Malmö University Hospital, Lund University, SE-20502, Malmö, Sweden. ola.forslund@mikrobiol.mas.lu.se
Ly
Hoang
H
Higgins
Geoff
G
D001315:Australia; D001706:Biopsy; D002280:Basal Cell Carcinoma / Q000175:diagnosis / Q000821:virology*; D002294:Carcinoma, Squamous Cell / Q000175:diagnosis / Q000821:virology*; D004279:DNA, Viral / Q000032:analysis*; D006801:Humans; D007642:Keratosis / Q000175:diagnosis / Q000821:virology; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis / Q000821:virology; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods; D012680:Sensitivity and Specificity; D012878:Skin Neoplasms / Q000175:diagnosis / Q000821:virology*; D014412:Tumor Virus Infections / Q000175:diagnosis / Q000821:virology
D004279:DNA, Viral