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Molecular detection of Tritrichomonas foetus in bovine samples: a novel real-time polymerase chain reaction (PCR) assay targeting EF1-alpha-Tf1 and a comparative study of published PCR techniques

Parasitology Research publish this investigation article

June 1st, 2022

The parasite T. foetus causes trichomonosis in cattle but is generally asymptomatic in males. Thus, many bulls carrying the disease go unnoticed, making the detection of T. foetus in bulls an important aspect for its control. Due to drawbacks posed by its cultivation, PCR is a preferred option for diagnostic laboratories. Most published PCR protocols target the genomic region compring the 18S, 5.8S, and 28S rRNA genes and internal transcribed spacers 1 and 2 (rRNA-ITS region), homologous to that of other Tritrichomonas species. There is minimal information on alternative genetic targets and no comparative studies have been published. We compared a protocol based on the microsatellite TfRE (called H94) and five protocols based on the rRNA-ITS region (called M06, M15, G02, G05, and N02). We also designed and evaluated a novel PCR-based assay on the EF1-alpha-Tf1 gene (called V21). The analytical sensitivity and specificity assays for the PCR protocols were performed according to the World Organisation for Animal Health (OIE) directives and the comparative study was performed with a widely used PCR (M06) on clinical samples from 466 breeding bulls. V21 showed a high degree of agreement with our reference M06 (kappa = 0.967), as well as M15 (kappa = 0.958), G05 (kappa = 0.948), and H94 (kappa = 0.986). Protocols H94 and V21 appear to be good approaches for confirming clinical cases in preputial bull samples when genomic regions alternative to rRNA-ITS are required. By contrast, N02 gave false negatives and G02 false positives




Polo C., Garcia-Seco T., Fernandez V., Hernandez M., Briones V., Diez-Guerrier A., Dominguez L. and Perez-Sancho M..




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Molecular detection of Tritrichomonas foetus in bovine samples: a novel real-time polymerase chain reaction (PCR) assay targeting EF1-alpha-Tf1 and a comparative study of published PCR techniques

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Molecular detection of Tritrichomonas foetus in bovine samples: a novel real-time polymerase chain reaction (PCR) assay targeting EF1-alpha-Tf1 and a comparative study of published PCR techniques



Participants:

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..

Zootecnia Análisis Clínicos VeterinariosZootecnia Análisis Clínicos Veterinarios.

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

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







Parasitology Research
FACTOR YEAR Q
2.289 2020

PMID: 35348895

ISSN: 09-0113



TITLE: Molecular detection of Tritrichomonas foetus in bovine samples: a novel real-time polymerase chain reaction (PCR) assay targeting EF1-alpha-Tf1 and a comparative study of published PCR techniques


JOURNAL: Parasitol. Res.


AÑO: 2022


PUBLISHER: Springer


AUTHORS: Polo C., Garcia-Seco T., Fernandez V., Hernandez M., Briones V., Diez-Guerrier A., Dominguez L. and Perez-Sancho M..


VISAVET PARTICIPANTS


First
Coral Polo Vaquero
2nd
Teresa García-Seco Romero
5th
Víctor Briones Dieste
6th
Alberto Antoine Díez Guerrier
7th
Lucas Domínguez Rodríguez
Last
Marta Pérez Sancho

DOI: https://doi.org/10.1007/s00436-022-07487-7


CITE THIS PUBLICATION:

Polo C., Garcia-Seco T., Fernandez V., Hernandez M., Briones V., Diez-Guerrier A., Dominguez L. and Perez-Sancho M. Molecular detection of Tritrichomonas foetus in bovine samples: a novel real-time polymerase chain reaction (PCR) assay targeting EF1-alpha-Tf1 and a comparative study of published PCR techniques. Parasitology Research. 2022. (A). ISSN: 09-0113. DOI: 10.1007/s00436-022-07487-7


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