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Detection and quantification of bovine papillomavirus DNA by digital droplet PCR in sheep blood
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  • Sante Roperto,
  • Anna Cutarelli,
  • Federica Corrado,
  • Francesca De Falco,
  • Canio Buonavoglia
Sante Roperto
Department of Veterinary Medicine
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Anna Cutarelli
Istituto Zooprofilattico Sperimentale del Mezzogiorno
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Federica Corrado
Istituto Zooprofilattico Sperimentale del Mezzogiorno
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Francesca De Falco
Universita degli Studi di Napoli Federico II Dipartimento di Medicina Veterinaria e Produzioni Animali
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Canio Buonavoglia
University of Bari
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Abstract

Abstract – Highly pathogenic bovine Delta papillomaviruses (δPVs) were detected and quantified for the first time using digital droplet polymerase chain reaction (ddPCR) by liquid biopsy in 103 clinically healthy sheep. Overall, ddPCR detected bovine δPVs in 68 blood samples (66%). Bovine papillomavirus (BPV) infection by a single genotype was revealed in 59% of the blood samples, and BPV coinfection by double, triple or quadruple genotypes was observed in 41% of liquid biopsies. The BPV-2 genotype was most frequently seen in sheep, whereas BPV-1 was the least common. Furthermore, ddPCR was very useful for detection and quantification; the BPV-14 genotype was observed for the first time in ovine species, displaying the highest prevalence in some geographical areas (Apulia). In 42 of the positive samples (61.8%), a single BPV infection was observed, 26 of which were caused by BPV-2 (61.9%) and 7 by BPV-13 (16.7%). BPV-14 was responsible for 7 single infections (16.7%) and BPV-1 for 2 single infections (4.7%). Multiple BPV coinfections were observed in the remaining 26 positive samples (38.2%), with dual BPV-2/BPV-13 infection being the most prevalent (84.6%). BPV infection by triple and quadruple genotypes was also observed in 11.5% and 3.8% of cases, respectively. The present study showed that ddPCR, a biotechnological refinement of conventional PCR, is by far the most sensitive and accurate assay for BPV detection. Therefore, ddPCR displayed diagnostic and epidemiological value resulting in the identification of otherwise undetectable BPV genotypes as well as their geographical distributions and suggesting that animal husbandry practices contribute to cross-species transmission of BPVs.
Dec 2021Published in Scientific Reports volume 11 issue 1. 10.1038/s41598-021-89782-4