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Evaluation of PCR assays for Campylobacter fetus detection and discrimination between C. fetus subspecies in bovine preputial wash samples

Artículo de investigación publicado en Theriogenology

15 de septiembre de 2021

Campylobacter fetus is a zoonotic pathogen found in cattle, in which it is one of the main causes of infectious infertility. Most diagnostic laboratories use PCR as quick easy tool for C. fetus identification. However, there is no standardized PCR assay for C. fetus detection and subspecies differentiation, hindering the comparison of results. In this study, we evaluated selected PCR assays targeting the 16S rRNA, gyrB, cpn60, cstA, cdtB and nahE genes for C. fetus identification and ISCfe1, sapB2, parA and virB11 for subspecies differentiation. Analytical sensitivity and specificity were assessed for each PCR assay, and the assays were then tested on 289 bull preputial samples that had also been analysed by 16S rRNA barcode metagenomics. In total, 41 C. fetus-positive samples were included. The P12 PCR assay targeting the gyrB gene performed best, detecting the pathogen in 95.1% of positive samples. For the discrimination of C. fetus subspecies, we were able to identify a proportion (85.4%) of the C. fetus-positive samples correctly as C. fetus venerealis with at least one subspecies-specific PCR, but C. fetus fetus was not detected in any of the samples tested. Remarkably, C. fetus subspecies amplification was observed following PCR on some samples (33.1%) considered C. fetus-negative, highlighting the need for rigorous criteria for discriminating between C. fetus subspecies, to improve understanding of the role of the two C. fetus subspecies in the epidemiology and pathogenesis of bovine infectious infertility




Polo Vaquero C., Garcia-Seco T., Hernandez M., Fernandez V., Rodriguez-Lazaro D., Goyache J., Dominguez L. y Perez-Sancho M..




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Evaluation of PCR assays for Campylobacter fetus detection and discrimination between C. fetus subspecies in bovine preputial wash samples

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Evaluation of PCR assays for Campylobacter fetus detection and discrimination between C. fetus subspecies in bovine preputial wash samples


Participantes:

Universidad ComplutenseServicio de Identificación y Caracterización Microbiana (ICM). Centro de Vigilancia Sanitaria Veterinaria (VISAVET). Universidad Complutense (UCM).

MAEVA SERVET, S.L.MAEVA SERVET, S.L..

Instituto Tecnológico Agrario de Castilla y LeónLaboratorio de Biología Molecular y Microbiología. Instituto Tecnológico Agrario de Castilla y León (ITACyL).

Zootecnia Análisis Clínicos Veterinarios.

Universidad de BurgosUnidad de Microbiología. Departamento de Biotecnología y Ciencia de los Alimentos. Universidad de Burgos (UBU).

Universidad ComplutenseDepartamento de Sanidad Animal. Facultad de Veterinaria. Universidad Complutense (UCM).







FACTOR YEAR Q
2.740 2020

NLMID: 421510

PMID: 34311221

ISSN: 0093-691X



TÍTULO: Evaluation of PCR assays for Campylobacter fetus detection and discrimination between C. fetus subspecies in bovine preputial wash samples


REVISTA: Theriogenology


NUMERACIÓN: 172:300-306


AÑO: 2021


EDITORIAL: Los Altos, Calif., Geron-X


AUTORES: Polo Vaquero C., Garcia-Seco T., Hernandez M., Fernandez V., Rodriguez-Lazaro D., Goyache J., Dominguez L. and Perez-Sancho M..


PARTICIPANTES VISAVET


2nd
Teresa García-Seco Romero
6th
Joaquín Goyache Goñi
7th
Lucas Domínguez Rodríguez
Last
Marta Pérez Sancho

DOI: https://doi.org/10.1016/j.theriogenology.2021.06.020


CITA ESTA PUBLICACIÓN:

Polo Vaquero C., Garcia-Seco T., Hernandez M., Fernandez V., Rodriguez-Lazaro D., Goyache J., Dominguez L. y Perez-Sancho M. Evaluation of PCR assays for Campylobacter fetus detection and discrimination between C. fetus subspecies in bovine preputial wash samples. Theriogenology. 172:300-306. 2021. (A). ISSN: 0093-691X. DOI: 10.1016/j.theriogenology.2021.06.020


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