ArticleApplied and environmental microbiology2005
Development of an F57 sequence-based real-time PCR assay for detection of Mycobacterium avium subsp. paratuberculosis in milk.
Article in Applied and environmental microbiology, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
- Comparative diagnostic evaluation and molecular typing of Mycobacterium avium subsp. Paratuberculosis in dromedary camels of Rajasthan, India.Veterinary research communications · 2025Article
- MAP, Johne's disease and the microbiome; current knowledge and future considerations.Animal microbiome · 2021Review
- Alpha-2-Macroglobulin as a New Promising Biomarker Improving the Diagnostic Sensitivity of Bovine Paratuberculosis.Frontiers in veterinary science · 2021Article
- Presence of Infection byMicroorganisms · 2020Article
- Diagnostic Sequences That DistinguishFrontiers in veterinary science · 2020Article
- A pilot study using environmental screening to determine the prevalence ofIrish veterinary journal · 2020Article
- Article
- Morphological and Molecular Characterization of a NewFrontiers in veterinary science · 2019Article
- Estimation of Performance Characteristics of Analytical Methods forFrontiers in microbiology · 2019Article
- EnvironmentalFrontiers in cellular and infection microbiology · 2018Article
- Comparison among the Quantification of Bacterial Pathogens by qPCR, dPCR, and Cultural Methods.Frontiers in microbiology · 2017Article
- Evaluation of a Commercial Real-Time PCR Kit for the Detection of Mycobacterium avium subspecies paratuberculosis in Milk.Current microbiology · 2016Article
- A novel mechanism for direct real-time polymerase chain reaction that does not require DNA isolation from prokaryotic cells.Scientific reports · 2016Article
- What Role Does Mycobacterium avium subsp. paratuberculosis Play in Crohn's Disease?Current infectious disease reports · 2015Article
- Comparison of fecal culture and F57 real-time polymerase chain reaction for the detection of Mycobacterium avium subspecies paratuberculosis in Swiss cattle herds with a history of paratuberculosis.Acta veterinaria Scandinavica · 2014Article
- Composite testing for ante-mortem diagnosis of Johne's disease in farmed New Zealand deer: correlations between bacteriological culture, histopathology, serological reactivity and faecal shedding as determined by quantitative PCR.BMC veterinary research · 2013Article
- Mycobacterium avium subspecies paratuberculosis: an insidious problem for the ruminant industry.Tropical animal health and production · 2013Review
- Enumeration of Mycobacterium avium subsp. paratuberculosis by quantitative real-time PCR, culture on solid media and optical densitometry.BMC research notes · 2012Article
- Highly specific and quick detection of Mycobacterium avium subsp. paratuberculosis in feces and gut tissue of cattle and humans by multiple real-time PCR assays.Journal of clinical microbiology · 2011Article
- A novel real-time PCR assay for specific detection and quantification of Mycobacterium avium subsp. paratuberculosis in milk with the inherent possibility of differentiation between viable and dead cells.BMC research notes · 2010Article
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Authors and funding
2 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A light cycler-based real-time PCR (LC-PCR) assay that amplifies the F57 sequence of Mycobacterium avium subsp. paratuberculosis was developed. This assay also includes an internal amplification control template to monitor the amplification conditions in each reaction. The targeted F57 sequence element is unique for M.avium subsp. paratuberculosis and is not known to exist in any other bacterial species. The assay specificity was demonstrated by evaluation of 10 known M. avium subsp. paratuberculosis isolates and 33 other bacterial strains. The LC-PCR assay has a broad linear range (2 x 10(1) to 2 x10(6) copies) for quantitative estimation of the number of M. avium subsp. paratuberculosis F57 target copies in positive samples. To maximize the assay's detection sensitivity, an efficient strategy for isolation of M. avium subsp. paratuberculosis DNA from spiked milk samples was also developed. The integrated procedure combining optimal M. avium subsp. paratuberculosis DNA isolation and real-time PCR detection had a reproducible detection limit of about 10 M. avium subsp. paratuberculosis cells per ml when a starting sample volume of 10 ml of M. avium subsp. paratuberculosis-spiked milk was analyzed. The entire process can be completed within a single working day and is suitable for routine monitoring of milk samples for M. avium subsp. paratuberculosis contamination. The applicability of this protocol for naturally contaminated milk was also demonstrated using milk samples from symptomatic M. avium subsp. paratuberculosis-infected cows, as well as pooled samples from a dairy herd with a confirmed history of paratuberculosis.
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