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Volume 16
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10.3390/v16101558
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Molecular Detection by Rolling Circle Amplification Combined with Deep Sequencing of Mixed Infection by Bovine Papillomaviruses 2 and 4 in Carcinoma In Situ of the Bovine Esophageal Mucosa

by
Bruna F. Matias
1,2,†,
Michele Lunardi
1,2,3,†,
Kátia C. B. Gonçalves
4,
Laurival A. Vilas-Boas
4,
Emanuele Gustani-Buss
5,
Ana Paula F. R. L. Bracarense
6,
Luiz Fernando C. Cunha Filho
3,
Alice F. Alfieri
1,2 and
Amauri A. Alfieri
1,7,*
1
Laboratory of Animal Virology, Department of Veterinary Preventive Medicine, Universidade Estadual de Londrina, Londrina 86057-970, Brazil
2
Multi-User Animal Health Laboratory, Molecular Biology Unit, Department of Veterinary Preventive Medicine, Universidade Estadual de Londrina, Londrina 86057-970, Brazil
3
Post Graduate Program in Animal Health and Production, Department of Agrarian Sciences, University Pitagoras Unopar, Arapongas 86702-670, Brazil
4
Department of General Biology, Universidade Estadual de Londrina, Londrina 86057-970, Brazil
5
Department of Microbiology, Immunology and Transplantation, Rega Institute, KU Leuven-University of Leuven, Box 1030, 3000 Leuven, Belgium
6
Laboratory of Animal Pathology, Department of Veterinary Preventive Medicine, Universidade Estadual de Londrina, Londrina 86057-970, Brazil
7
National Institute of Science and Technology for Dairy Production Chain (INCT–LEITE), Universidade Estadual de Londrina, Londrina 86057-970, Brazil
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Viruses 2024, 16(10), 1558; https://doi.org/10.3390/v16101558
Submission received: 6 August 2024 / Revised: 24 September 2024 / Accepted: 29 September 2024 / Published: 30 September 2024
(This article belongs to the Special Issue Animal Papillomaviruses Research)
Versions Notes

Abstract

Papillomaviruses (PVs) are oncogenic and infect the skin and mucosa of various host species. Considering the recent advances in research on PVs using rolling circle amplification (RCA) followed by high-throughput sequencing (HTS), in this study, we aimed to investigate the bovine papillomavirus (BPV) types associated with proliferative lesions in the upper alimentary tract of an affected bull and characterize the viral strains through complete genome sequencing using this strategy. We analyzed the PV strains associated with two hyperplastic esophageal lesions through PCR using degenerate primer pairs and RCA, followed by HTS. HTS of the libraries generated using RCA products provided the whole genome sequence of BPV4 present in squamous papilloma, whereas the complete genome sequence of BPV2 and subgenomic fragments of BPV4 were identified in carcinoma in situ (CIS). For the first time, we have sequenced BPV2 identified from the CIS of the bovine upper alimentary canal. Additionally, RCA followed by HTS allowed characterization of the mixed infection by BPV2 and BPV4 in this lesion. These data reveal that BPV4 is not the only BPV type present in CIS of the esophageal mucous membrane; moreover, a mixed infection caused by BPV2 and BPV4 at the tested anatomical site was demonstrated.
Keywords: cattle; upper alimentary tract; esophagus; papilloma; histopathology; coinfection; polymerase chain reaction; next generation sequencing cattle; upper alimentary tract; esophagus; papilloma; histopathology; coinfection; polymerase chain reaction; next generation sequencing

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MDPI and ACS Style

Matias, B.F.; Lunardi, M.; Gonçalves, K.C.B.; Vilas-Boas, L.A.; Gustani-Buss, E.; Bracarense, A.P.F.R.L.; Filho, L.F.C.C.; Alfieri, A.F.; Alfieri, A.A. Molecular Detection by Rolling Circle Amplification Combined with Deep Sequencing of Mixed Infection by Bovine Papillomaviruses 2 and 4 in Carcinoma In Situ of the Bovine Esophageal Mucosa. Viruses 2024, 16, 1558. https://doi.org/10.3390/v16101558

AMA Style

Matias BF, Lunardi M, Gonçalves KCB, Vilas-Boas LA, Gustani-Buss E, Bracarense APFRL, Filho LFCC, Alfieri AF, Alfieri AA. Molecular Detection by Rolling Circle Amplification Combined with Deep Sequencing of Mixed Infection by Bovine Papillomaviruses 2 and 4 in Carcinoma In Situ of the Bovine Esophageal Mucosa. Viruses. 2024; 16(10):1558. https://doi.org/10.3390/v16101558

Chicago/Turabian Style

Matias, Bruna F., Michele Lunardi, Kátia C. B. Gonçalves, Laurival A. Vilas-Boas, Emanuele Gustani-Buss, Ana Paula F. R. L. Bracarense, Luiz Fernando C. Cunha Filho, Alice F. Alfieri, and Amauri A. Alfieri. 2024. "Molecular Detection by Rolling Circle Amplification Combined with Deep Sequencing of Mixed Infection by Bovine Papillomaviruses 2 and 4 in Carcinoma In Situ of the Bovine Esophageal Mucosa" Viruses 16, no. 10: 1558. https://doi.org/10.3390/v16101558

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