Comparison of two PCR strategies for the detection of bovine papillomavirus

Abstract

Bovine papillomavirus (BPV) is a diverse group of double-stranded DNA oncogenic viruses, which have been detected in epithelial lesions and body fluids. Most studies of BPV infection rely on a single method for DNA detection; however the use of any single method or technique may underestimate the true prevalence of this virus. The purpose of this study was to compare two PCR strategies for the detection of BPV in skin lesions and fluids: these involve the use of BPV type-specific and consensus primers. Seventy-two cutaneous lesions, 57 blood samples and 59 semen samples were collected. PCR was used with the FAP consensus primers and BPV type-specific primers (for BPVs 2, 3, 4, 5, 8, 9 and 10), along with sequencing assays, to detect the BPV types. Phylogenetic analysis was carried out by means of the maximum likelihood method. It was found that both FAP and BPV type-specific primer sets could amplify BPV types of DNA in skin lesions, blood and semen samples. However, the BPV type-specific primers were more sensitive than the consensus primers and were able to detect co-infection of BPV in the samples. The consensus primers amplified five BPV types and were more suitable for detecting new putative BPV types. Thus, account should be taken of both PCR primer systems to identify co-infection, the presence of novel viruses, and avoid false-negative results.

Molecular identification of bovine papillomaviruses in dairy and beef cattle: first description of Xi- and Epsilonpapillomavirus in Turkey

“…A recent study described molecular identification of BPV-1 in cattle with cutaneous papillomatosis in Turkey, but BPV-2 was not detected (20). In the present study, the detection rate of BPV types in papilloma-like samples using BPV type-specific primers was greater than that for FAP59/64 degenerate primers, like the results described by Silva et al (16). Furthermore, 7 BPV types, including genera Deltapapillomavirus (BPVs 1 and 2), Xipapillomavirus (BPVs 3, 4, 6, and 9), and Epsilonpapillomavirus (BPV-8), were identified.…”

“…The L1 gene of papillomavirus has been used to detect BPVs by PCR using the degenerated FAP59/64 primer pair, which is the most conserved region (11,15). Nevertheless, the use of degenerated FAP59/64 primers together with type-specific primers is recommended (16,17), since the degenerated FAP59/64 primers have a low sensitivity for detecting new papillomavirus types and are unsuccessful for detecting some BPV types such as BPV-4 and BPV-9 (17).…”

Genetic characterization of Amazonian bovine papillomavirus reveals the existence of four new putative types

“…Although some BPVs have higher affinities to specific primers [23], in the present study the primer pair FAP59/64 was able to amplify bovine papillomavirus DNA from all known BPVs genera. Despite the lower level of sensitivity of these primers when compared to specific primers [23] they have been used in many studies, being an important tool to enlarge the knowledge of BPVs diversity and enabling the discovery of a broad range of novel BPVs [10][11][12][13]16].…”

Papillomaviruses: a systematic review

“…Studies about BPV diversity and prevalence are mandatory to develop novel therapeutic methods (Silva et al , 2013b), since immunity is species-specific (Claus et al. , 2007).…”