Development of a novel PCR-RFLP assay for improved detection and typing of bovine papillomaviruses

Journal of Virological Methods
Kyoko KawauchiShinichi Hatama

Abstract

A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay was developed to detect and type bovine papillomaviruses (BPVs) from tumors in cattle. Two degenerate primer sets targeting the BPV L1 gene, subAup/subAdw and subBup/subBdw, and one restriction enzyme RsaI were used in this assay. In silico analyses of the restriction enzyme sites in the PCR fragments of 13 BPV sequences (BPV-1 to -13) revealed that all known BPVs are differentiated by the PCR-RFLP assay. Analyses of 63 previously typed clinical samples, that included teat papillomas and both esophageal and urinary bladder cancer biopsies, show that the assay clearly differentiates between eight clinically important BPV types (BPV-1 to -6, -9, -10), and discriminates between single and multiple infections. To further assess the reliability of the PCR-RFLP method amplified fragments were sequenced. A high correlation (95%) was observed when the results of the PCR-RFLP method were compared with PCR-sequencing. Differences in typing occurred for 3 of 63 specimens; PCR-RFLP identified additional BPV types in these specimens, while the PCR-sequencing identified only one. These results indicate that the PCR-RFLP method reported here is simpler and m...Continue Reading

References

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Citations

Feb 18, 2017·Genetics and Molecular Biology·Rodrigo Pinheiro AraldiWilly Beçak
Apr 1, 2018·Transboundary and Emerging Diseases·C DaudtC W Canal
Sep 14, 2018·The Journal of Veterinary Medical Science·Hidekatsu ShimakuraTakeshi Haga
Jun 4, 2019·The Journal of Veterinary Medical Science·Nanako Yamashita-KawanishiTakeshi Haga
Aug 31, 2018·The Journal of Veterinary Medical Science·Shinichi HatamaKoichi Kadota
Jul 9, 2020·Virus Genes·Nanako Yamashita-KawanishiTakeshi Haga
Dec 11, 2020·Equine Veterinary Journal·Nanako Yamashita-KawanishiTakeshi Haga
Dec 3, 2020·Pathogens·Nanako Yamashita-Kawanishi, Takeshi Haga

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