Comparison of culture and a novel 5' Taq nuclease assay for the direct detection of Campylobacter fetus subspecies venerealis in clinical specimens from cattle

McMillen, Lyle, Fordyce, Geoffry, Doogan, Vivienne J. and Lew, Ala E. (2006) Comparison of culture and a novel 5' Taq nuclease assay for the direct detection of Campylobacter fetus subspecies venerealis in clinical specimens from cattle. Journal of Clinical Microbiology, 44 3: 938-945. doi:10.1128/JCM.44.3.938-945.2006

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Author McMillen, Lyle
Fordyce, Geoffry
Doogan, Vivienne J.
Lew, Ala E.
Title Comparison of culture and a novel 5' Taq nuclease assay for the direct detection of Campylobacter fetus subspecies venerealis in clinical specimens from cattle
Formatted title
Comparison of culture and a novel 5' Taq nuclease assay for the direct detection of Campylobacter fetus subspecies venerealis in clinical specimens from cattle
Journal name Journal of Clinical Microbiology   Check publisher's open access policy
ISSN 0095-1137
1098-660X
Publication date 2006-03
Sub-type Article (original research)
DOI 10.1128/JCM.44.3.938-945.2006
Open Access Status File (Publisher version)
Volume 44
Issue 3
Start page 938
End page 945
Total pages 8
Place of publication Washington, DC, United States
Publisher American Society for Microbiology
Language eng
Formatted abstract
A Campylobacter fetus subsp. venerealis-specific 5' Taq nuclease PCR assay using a 3' minor groove binder-DNA probe (TaqMan MGB) was developed based on a subspecies-specific fragment of unknown identity (S. Hum, K. Quinn, J. Brunner, and S. L. On, Aust. Vet. J. 75:827-831, 1997). The assay specifically detected four C. fetus subsp. venerealis strains with no observed cross-reaction with C. fetus subsp. fetus-related Campylobacter species or other bovine venereal microflora. The 5' Taq nuclease assay detected approximately one single cell compared to 100 and 10 cells in the conventional PCR assay and 2,500 and 25,000 cells from selective culture from inoculated smegma and mucus, respectively. The respective detection limits following the enrichments from smegma and mucus were 5,000 and 50 cells/inoculum for the conventional PCR compared to 500 and 50 cells/inoculum for the 5' Taq nuclease assay. Field sampling confirmed the sensitivity and the specificity of the 5' Taq nuclease assay by detecting an additional 40 bulls that were not detected by culture. Urine-inoculated samples demonstrated comparable detection of C. fetus subsp. venerealis by both culture and the 5' Taq nuclease assay; however, urine was found to be less effective than smegma for bull sampling. Three infected bulls were tested repetitively to compare sampling tools, and the bull rasper proved to be the most suitable, as evidenced by the improved ease of specimen collection and the consistent detection of higher levels of C. fetus subsp. venerealis. The 5' Taq nuclease assay demonstrates a statistically significant association with culture (χ2 = 29.8; P < 0.001) and significant improvements for the detection of C. fetus subsp. venerealis-infected animals from crude clinical extracts following prolonged transport.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Queensland Alliance for Agriculture and Food Innovation
ERA 2012 Admin Only
 
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Created: Thu, 13 Oct 2011, 10:38:24 EST by Dr Alicja Lew-tabor on behalf of Qld Alliance for Agriculture and Food Innovation