Human papillomavirus can be detected by amplification of viral DNA. A novel one-step PCR (PM-PCR) was evaluated for amplification of a 117-bp fragment from the E1 region. It permitted ultrasensitive detection of all 25 known human papillomavirus genotypes from the beta-papillomavirus genus. The intra-and intertypic sequence variations of the 77-bp interprimer region were studied. Genotype-specific probes as well as general probes were selected for the 25 established beta-papillomavirus types, and a reverse hybridization assay (RHA) was developed (PM-PCR RHA method). The analytical sensitivity of the PM-PCR RHA method was 10 to 100 viral genomes. The one-step PM-PCR turned out to be more sensitive than the previously described nested MaHa-PCR for beta-papillomavirus detection. The PM-PCR RHA method was able to detect and identify beta-papillomavirus types in frozen patient material as well as in poorly amplifiable material such as formalinfixed, paraffin-embedded skin biopsy specimens. Inter-and intralaboratory variability experiments showed that the reproducibility of the assay was very high. In conclusion, the one-step PM-PCR together with the RHA allows extremely sensitive, specific, and reproducible detection of beta-papillomavirus DNA as well as reliable identification of beta-papillomavirus genotypes in both fresh and paraffin-embedded patient material.Papillomaviruses (PV) constitute a group of viruses associated with benign and malignant lesions of cutaneous and mucosal epithelia. So far, more than 100 different PV genotypes have been identified, of which approximately 48 types have been detected in human cutaneous lesions (12). These include the beta-papillomavirus (beta-PV) genus comprising the human papillomavirus (HPV) types 5,8,9,12,14,15,17,19,20,21,22,23,24,25, 36, 37, 38, 47, 49, 75, 76, 80, and 93 and candidate types 92 (cand92) and cand96. Based on partial sequences, however, probably more than 35 new types have to be added to the 25 known beta-PV types (19). Originally, types of the beta genus have been found in skin lesions from patients with the rare hereditary disease epidermodysplasia verruciformis. These patients develop flat cutaneous warts and macular lesions. They arise early in life and have a high chance to progress into squamous cell carcinoma (SCC) on sun-exposed sites. In these SCCs, mostly HPV types 5 and 8 have been detected, suggesting that these types are high-risk HPV types (19).DNA from beta-PV types was identified mainly by nested PCR in 30 to 50% of SCCs in immunocompetent patients and in up to 90% of the SCCs in immunosuppressed patients, e.g., renal transplant recipients (19). The high prevalence of beta-PV types in these SCCs and their precursor lesions (solar keratoses) suggests an involvement in the carcinogenesis. Recent epidemiological case control studies have further corroborated this hypothesis by showing that the presence of beta-PV DNA in eyebrow hairs was associated with a history of cutaneous solar keratoses (7) and cutaneous SCC (23).Little is known about the...

Background: Beta-human papillomavirus (betaPV) may play a role in the development of cutaneous squamous cell carcinoma (SCC). However betaPV is highly prevalent, and it may only be people with a higher viral load who have increased risk of SCCs. We therefore examined the association between betaPV load and SCCs.Methods: We recruited 448 immunocompetent cases with SCCs and 464 controls from Italy and Australia and 497 immunosuppressed organ transplant recipients (OTR; 179 cases and 318 controls) from Europe. We used reverse hybridization to genotype 25 betaPV types in eyebrow hair follicles and determined the viral load for eight selected types using quantitative PCR. We used logistic regression to assess associations between type-specific and cumulative viral load and SCCs.Results: Australian and OTR participants in the highest cumulative load tertile were at significantly higher risk of SCCs than those in the lowest tertile. Those with more than four betaPV types in the high load tertile were at approximately three-fold increased risk of SCCs. In Australia, HPV23 and 36 loads were significantly associated with SCCs, with borderline associations for HPV5 and 38. In OTR, HPV8 and 38 loads were significantly associated and HPV20 and 36 were borderline. We found little evidence for an association between load and SCCs in Italy.Conclusions: High viral load may be associated with risk of cutaneous SCCs, with total load seemingly more important than the load of any specific type.Impact: Our findings lend weight to the hypothesis that HPV plays a role in skin carcinogenesis. Cancer Epidemiol Biomarkers Prev; 22(4); 719-27. Ó2013 AACR.

BackgroundCervical cancer incidence and mortality rates in Sub-Saharan Africa (SSA) remain high due to several factors including low levels of uptake of cervical cancer screening. Self-collection of cervicovaginal samples for HPV DNA testing may be an effective modality that can increase uptake of cervical cancer screening in SSA and hard to reach populations in developed countries. We investigated whether self-collection of cervicovaginal samples for HPV DNA tests would be associated with increased uptake of screening compared with clinic based collection of samples. Furthermore, we compared the quality of samples collected by both approaches for use in HPV genotyping.MethodsWe conducted a community based randomized trial in a semi-urban district of Abuja, Nigeria with 400 women, aged 30 to 65 years randomized to either hospital-collection or self-collection of cervicovaginal samples. We compared cervical cancer screening uptake among the 2 groups and evaluated the concentration of human DNA in the samples by measuring RNase P gene levels using qPCR. High-risk HPV DNA detection and typing was done using the GP5+/6+ Luminex system.ResultsMost participants in the self-collection arm (93%, 185/200) submitted their samples while only 56% (113/200) of those invited to the hospital for sample collection attended and were screened during the study period (p value < 0.001). Human genomic DNA was detected in all but five (1.7%) participants, all of whom were in the self-collection arm. The prevalence of high-risk HPV in the study population was 10% with types 35, 52 and 18 being the commonest.ConclusionsOur study shows that self-sampling significantly increased uptake of HPV DNA based test for cervical cancer screening in this population and the samples collected were adequate for HPV detection and genotyping. Cervical cancer screening programs that incorporate self-sampling and HPV DNA tests are feasible and may significantly improve uptake of cervical cancer screening in SSA.

Similar CIN2+ sensitivity for HPV testing in first-void urine, physician-taken smear and brush-based self-sample.