RT Journal Article
T1 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
A1 Polo, Coral
A1 Fernández, Víctor
A1 Hernández, Marta
A1 Briones Dieste, Víctor
A1 Díez Guerrier, Alberto Antoine
A1 Pérez Sancho, Marta
A1 Domínguez Rodríguez, Lucas José
A1 García-Seco Romero, María Teresa
AB 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.
PB Elsevier
SN 0932-0113
YR 2022
FD 2022-03-29
LK https://hdl.handle.net/20.500.14352/71478
UL https://hdl.handle.net/20.500.14352/71478
LA eng
NO CRUE-CSIC (Acuerdos Transformativos 2022)
NO Comunidad de Madrid
NO R&D Agreement between the Instituto Tecnológico Agrario de Castilla y León, VISAVET-Universidad Complutense de Madrid, and Universidad de Burgos
DS Docta Complutense
RD 6 may 2024