Improved detection of Tritrichomonas foetus in bovine diagnostic specimens using a novel probe-based real time PCR assay

McMillen, L.; Lew, Ala E.

doi:10.1016/j.vetpar.2006.06.012

Cited by 55 publications

(48 citation statements)

References 54 publications

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“…A combination of measures will be needed, which should be & Lew (2006) devised to specifically fit individual cases. In order to achieve eradication, artificial insemination will have to be widely accepted by US ranchers.…”

Section: Discussionmentioning

confidence: 99%

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Diagnosis of Tritrichomonas foetus-infected bulls, an ultimate approach to eradicate bovine trichomoniasis in US cattle?

Yao

2013

Journal of Medical Microbiology

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Diagnosis ofBovine trichomoniasis is a sexually transmitted protozoan disease with a worldwide distribution. It has been endemic in the USA for more than 80 years. Mississippi and all the states west of the Mississippi River, except Iowa and Minnesota, have rules/regulations to reduce the spread of the disease. The core of these regulations consists of testing bulls and prohibiting importation of nonTritrichomonas foetus-free bulls. Factors such as sampling methods and intervals, shipping medium and temperature, and testing techniques are reviewed for their effect on diagnostic accuracy. Finally, a comprehensive approach for controlling and eventually eradicating the disease is presented.

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Section: Discussionmentioning

confidence: 99%

“…In addition to its specificity, gel-PCR is also very sensitive. It can detect as low as a single organism (Chen & Li, 2001;Felleisen et al, 1998;McMillen & Lew, 2006) (Table 3). Of 29 samples collected from five infected bulls with a resting period of 2-4 days, 24 (83 %) were positive (Mukhufhi et al, 2003).…”

Section: Gel-pcrmentioning

confidence: 99%

Diagnosis of Tritrichomonas foetus-infected bulls, an ultimate approach to eradicate bovine trichomoniasis in US cattle?

Yao

2013

Journal of Medical Microbiology

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“…Additionally as the organism dies in the culture media it degrades and the by-products of the degradation results in a breakdown of the organisms DNA [8,9].…”

Section: Methods To Detectmentioning

confidence: 99%

Tritrichomonas foetus in Beef Bulls

2014

J Veterinar Sci Technol

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Tritrichomonas foetus is a production and regulatory concern for beef producers in the Western United States and more recently in states of the Mississippi Valley. Traditionally preputial scrapings have been collected, cultured in enriched media, and examined microscopically. PCR techniques are now being used extensively to confirm culture results or as a stand-alone test for the organism. This technology offers the advantage of distinguishing the pathogenic Tritrichomonas foetus organism from other nonpathogenic fecal organisms.

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“…13 The laboratory included exogenous internal positive control reagents and DNA n in the qPCR reaction mixture. Laboratory F did not report on the baseline or analysis settings.…”

Section: Quantitative Pcr Methodsmentioning

confidence: 99%

Pooling of cultured samples and comparison of multistate laboratory workflows with the MagMAX sample preparation system and VetMAX quantitative polymerase chain reaction reagents for detection ofTritrichomonas foetus–colonized bulls

et al. 2013

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The objectives of the current study were 1) to compare sample preparation workflows and quantitative real-time polymerase chain reaction assays (qPCR) as currently used in veterinary diagnostic laboratories with a study protocol utilizing commercially available reagents for individual Tritrichomonas foetus testing, 2) to assess the accuracy of pooling cultured smegma samples followed by extraction and qPCR testing as used in the study laboratory, and 3) to assess the specificity of the currently used primers and probes by sequencing all positive and presumptive positive samples identified in the study laboratory in an attempt to capture any nucleotide variability between T. foetus isolates and to rule out false-positive results possibly due to Simplicimonas moskowitzi. Eight hundred three cultured smegma samples were collected from different regions of the United States with the collaboration of 5 veterinary testing laboratories. The samples were processed individually by the respective laboratories, and then sent to the study laboratory and retested using the study protocol. Comparison testing showed an overall agreement of 95.89% between the veterinary testing laboratories and the study laboratory. One hundred seventy-six positive or presumptive positive samples plus 625 negative qPCR samples were combined and retested using a pooling protocol. Pools consisted of 1 positive sample and 4 negative samples (1/5). These pools were processed using the same study laboratory protocols, and 96% of the positive samples were detected in these pools. Nested PCR followed by sequencing confirmed 175 of the 178 samples classified as positive or presumptive positive in the study laboratory as containing T. foetus–specific DNA.

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Improved detection of Tritrichomonas foetus in bovine diagnostic specimens using a novel probe-based real time PCR assay

Cited by 55 publications

References 54 publications

Diagnosis of Tritrichomonas foetus-infected bulls, an ultimate approach to eradicate bovine trichomoniasis in US cattle?

Diagnosis of Tritrichomonas foetus-infected bulls, an ultimate approach to eradicate bovine trichomoniasis in US cattle?

Tritrichomonas foetus in Beef Bulls

Pooling of cultured samples and comparison of multistate laboratory workflows with the MagMAX sample preparation system and VetMAX quantitative polymerase chain reaction reagents for detection ofTritrichomonas foetus–colonized bulls

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