A broad range of human papillomavirus types detected with a general PCR method suitable for analysis of cutaneous tumours and normal skin

Abstract

A pair of degenerate PCR primers (FAP59/64) was designed from two relatively conserved regions of the L1 open reading frame of most human papillomaviruses (HPV). The size of the generated amplicon was about 480 bp. PCR using these primers was found capable of amplifying DNA from 87 % (65/75) of the HPV types tested, its sensitivity being 1-10 copies for HPV-5, -20 and -30 clones. HPV was found in 63 % (5/8) of tumour samples and in 63 % (5/8) of normal skin biopsies from patients with various cutaneous tumours. HPV-5, HPV-8, HPV-12, HPVvs20-4 and six putatively novel HPV types were identified. No correlation was found to exist between specific HPV and tumour types. Skin surface swab samples from one or more sites on three of four healthy volunteers were found to contain HPV, types 12 and 49 being identified, as well as eight novel HPV types, two of which were also found among the patients. In all, HPV was detected in 75 % (9/12) of those tested, five HPV types and 12 novel candidate types being identified, and 37 % (7/19) of HPV-positive samples were found to manifest more than one HPV type. All the HPV detected manifested high degrees of nucleotide sequence similarity with HPV types associated with skin lesions and epidermodysplasia verruciformis. The overall HPV finding in the skin samples was 50 % (20/40) using the FAP primers as compared to 18 % (7/40) using another PCR test designed for skin types. The results thus suggest the new method to be sensitive and generally applicable for detecting cutaneous HPV.

Evaluating the role of human papillomaviruses in conjunctival neoplasia

“…Testing for HPV was carried out with generalprimer-mediated PCR using the following primer pairs: (a) MY09/ MY11 (Resnick et al, 1990) and et al, 1995), which preferentially amplify mucosal HPVs; (c) CP65/CP70 followed by CP66/CP69 for the amplification of EVrelated HPV types (Berkhout et al, 1995); (d) FAP59/FAP64 for the amplification of 75 different types of mucosal, cutaneous and EVrelated HPVs (Forslund et al, 1999). In addition, we used typespecific PCR targeting E6 genes of individual HPV16 and HPV38 types Caldeira et al, 2003).…”

Epidermodysplasia verruciformis human papillomavirus types and carcinoma of the conjunctiva: a pilot study

“…HPV detection was performed by means of polymerase chain reaction (PCR) assays using different sets of primers (Chen et al, 1994;Berkhout et al, 1995;Jacobs et al, 1995;Forslund et al, 1999, Harwood et al, 1999Caldeira et al, 2003). Laboratory personnel were blinded to the histological diagnosis of each patient.…”

Human papillomavirus infection and ultraviolet light exposure as epidermoid inclusion cyst risk factors in a patient with epidermodysplasia verruciformis?

“…Epidermodysplasia verruciformis (EV) is a rare disease characterized by extensive symptomatic cutaneous infection with specific human papillomavirus (HPV) genotypes including HPV 5,8,9,12,14,15,17,[19][20][21][22][23][24][25]46, 47, 50. 1 Two susceptibility loci for EV have been identified, EV1 on chromosome 17q, and EV2 on chromosome 2p.…”