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Digital droplet PCR for the detection and quantification of circulating bovine Deltapapillomavirus
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  • Francesca De Falco,
  • Federica Corrado,
  • Anna Cutarelli,
  • LEONARDO LEONARDI,
  • Sante Roperto
Francesca De Falco
Universita degli Studi di Napoli Federico II Dipartimento di Medicina Veterinaria e Produzioni Animali
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Federica Corrado
Istituto Zooprofilattico Sperimentale del Mezzogiorno
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Anna Cutarelli
Istituto Zooprofilattico Sperimentale del Mezzogiorno
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LEONARDO LEONARDI
University of Perugia- Faculty of Veterinary Medicine
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Sante Roperto
Department of Veterinary Medicine
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Abstract

In this study, the digital droplet polymerase chain reaction (ddPCR) was used to quantify circulating bovine papillomavirus (BPV; genus: Deltapapillomavirus) levels in blood samples from 25 healthy cows and 15 cows with chronic enzootic hematuria due to papillomavirus-associated bladder tumors. ddPCR detected the BPV DNA in 95% of all the samples (i.e., in 24 of the healthy cows and 14 of the diseased animals), whereas quantitative real time PCR (qPCR) detected it in only 57.5% of the same blood samples, with the percentage differences between ddPCR and qPCR being statistically significant (P value  0.05), according to the 2 test of Campbell and Richardson. Furthermore, ddPCR detected BPV infections by a single genotype and by multiple genotypes in 37% and 63% of the cows, respectively, whereas qPCR detected these in 16% and 16%, respectively. Of the two assays, ddPCR was the more sensitive and accurate clinical diagnostic tool, allowing the detection of otherwise undetectable BPV genotypes, and consequently, a higher number of BPV co-infections. qPCR failed to detect many BPV co-infections by multiple genotypes. Therefore, ddPCR may be an essential tool for improving diagnostic procedures, allowing the identification of the genotypic distribution of BPV and a better understanding about the territorial divergence, if any, of the BPV prevalence in different areas. No significant differences in the blood viral load estimations were observed between the two animal groups, suggesting that the bloodstream could be a site of primary infection. Finally, as BPV DNA was detected in cows affected by noninvasive urothelial tumors, including papilloma and papillary urothelial neoplasms of low malignant potential, the circulating BPVs appeared to be independent of the status of urothelial neoplasms. Therefore, unlike in humans, circulating BPVs cannot be an actual prognostic marker of urothelial tumors in cows.

Peer review status:Published

05 May 2020Submitted to Transboundary and Emerging Diseases
06 May 2020Submission Checks Completed
06 May 2020Assigned to Editor
13 May 2020Reviewer(s) Assigned
10 Jun 2020Review(s) Completed, Editorial Evaluation Pending
10 Jun 2020Editorial Decision: Revise Major
16 Jun 20201st Revision Received
17 Jun 2020Assigned to Editor
17 Jun 2020Submission Checks Completed
17 Jun 2020Reviewer(s) Assigned
03 Jul 2020Review(s) Completed, Editorial Evaluation Pending
03 Jul 2020Editorial Decision: Revise Major
15 Jul 20202nd Revision Received
15 Jul 2020Submission Checks Completed
15 Jul 2020Assigned to Editor
15 Jul 2020Reviewer(s) Assigned
07 Aug 2020Review(s) Completed, Editorial Evaluation Pending
07 Aug 2020Editorial Decision: Revise Minor
12 Aug 20203rd Revision Received
12 Aug 2020Assigned to Editor
12 Aug 2020Submission Checks Completed
12 Aug 2020Editorial Decision: Accept
16 Aug 2020Published in Transboundary and Emerging Diseases. 10.1111/tbed.13795