The Development of Novel Primer Sets to Specifically Amplify Each of the Five Different Deltapapillomaviruses That Cause Neoplasia after Cross-Species Infection.

Cíntia Daudt,Flavio Chaves Da Silva,Kristene Gedye,John S Munday

doi:10.3390/vetsci8100208

Cíntia Daudt, Flavio Chaves Da Silva + Show 2 more

Open Access

https://doi.org/10.3390/vetsci8100208

Copy DOI

Abstract

Bovine papillomavirus (BPV) types 1 and 2 are recognized as the main cause of equine sarcoids. However, some studies report that up to a quarter of these tumors do not contain detectible BPV1 or BPV2 DNA. The absence of detectible BPV1 or BPV2 in these sarcoids suggests the possible involvement of other papillomavirus types. Currently, five deltapapillomaviruses are recognized to cause mesenchymal neoplasia after cross-species infection. In addition to BPV1 and BPV2, BPV13 has been associated with equine sarcoids in Brazil, BPV14 has been associated with feline sarcoids, and Ovis aries papillomavirus 2 caused a sarcoid-like lesion in a pig. To investigate the cause of equine sarcoids, PCR primers were developed to specifically amplify each of the five different deltapapillomaviruses that have been associated with mesenchymal neoplasia. The specificity of these primers was confirmed using samples of formalin-fixed tissue known to contain each PV type. These primers allow rapid and sensitive detection of deltapapillomavirus DNA in equine sarcoids. As studies have revealed marked regional variability in the cause of equine sarcoids, these primers will be useful to determine the predominant PV type causing sarcoids in a region. Additionally, there is a single report describing mixed infections by BPV1 and BPV2 in equine sarcoids. The specific primer sets are expected to enable more sensitive detection of mixed infections in equine sarcoids. Determining the cause of equine sarcoids is important as vaccines are developed to prevent these common malignant neoplasms.

Highlights

Paraffin tissue blocks containing formalin-fixed samples of an equine sarcoid that contained BPV1 DNA, a bovine anal fibropapilloma containing BPV2 DNA [17], a feline sarcoid that contained BPV14 DNA, and a sarcoid-like lesion from the mouth of a pig known to contain OaPV2 DNA [12] were used in the study
Using the Primer-BLAST tool allowed the development of 5 primer sets to amplify DNA from each of the five deltapapillomavirus types that have been associated with sarcoids and sarcoid-like lesions (Table 1)
To ensure that the primers only amplified the targeted PV type and did not have a high affinity to the other deltapapillomavirus types, the primer sequences were compared to the published sequences of the other PV types (Table 2)

Summary

Introduction

Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Papillomaviruses (PVs) are double-stranded circular DNA viruses that infect most vertebrate species [1]. PVs are able to promote cell growth and replication, and PV infection can result in the development of a hyperplastic papilloma (wart) [2]. PVs can promote the development of irreversible changes within the cell DNA and the development of neoplasia [2]. PVs are classified using the highly conserved L1 gene.

Objectives

Methods

Results

Discussion

Conclusion