ArticleApplied and environmental microbiology2014
Development and evaluation of a novel multicopy-element-targeting triplex PCR for detection of Mycobacterium avium subsp. paratuberculosis in feces.
Article in Applied and environmental microbiology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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34 citing papers in PubMed, 52 citations in OpenAlex.
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- Evaluation of the reproducibility and performance characteristics of the Phagomagnetic separation-qPCR assay for rapidly detecting viableFrontiers in veterinary science · 2025Article
- Evaluation of the Effect of a Recent Comparative Intradermal Tuberculin Test on the Humoral Diagnosis of Paratuberculosis Using Serum and Milk Samples from Goats.Veterinary sciences · 2024Article
- Evaluation of alternative vaccination routes against paratuberculosis in goats.Frontiers in veterinary science · 2024Article
- Probiotic bacteria can modulate immune responses to paratuberculosis vaccination.Frontiers in cellular and infection microbiology · 2024Article
- Comparison of blood parameters according to fecal detection ofJournal of veterinary science · 2023Article
- Improved DNA Amplification of the Hallmark ISApplied and environmental microbiology · 2023Article
- Assessment of the frequency of Mycobacterium bovis shedding in the faeces of naturally and experimentally TB infected cattle.Journal of applied microbiology · 2022Article
- A rapid phage assay for detection of viable Mycobacterium avium subsp. paratuberculosis in milk.Scientific reports · 2022Article
- Oral vaccination stimulates neutrophil functionality and exerts protection in a Mycobacterium avium subsp. paratuberculosis infection model.NPJ vaccines · 2021Article
- Bovine Intelectin 2 Expression as a Biomarker of Paratuberculosis Disease Progression.Animals : an open access journal from MDPI · 2021Article
- Molecular characterisation of Mycobacterium avium subsp. paratuberculosis in Australia.BMC microbiology · 2021Article
- Influence of Heterologous and Homologous Vaccines, and Their Components, on the Host Immune Response and Protection Against Experimental Caprine Paratuberculosis.Frontiers in veterinary science · 2021Article
- Application of Volatilome Analysis to the Diagnosis of Mycobacteria Infection in Livestock.Frontiers in veterinary science · 2021Review
- Alpha-2-Macroglobulin as a New Promising Biomarker Improving the Diagnostic Sensitivity of Bovine Paratuberculosis.Frontiers in veterinary science · 2021Article
- Detection of Wood Mice (Microorganisms · 2020Article
- Alternative Vaccination Routes against Paratuberculosis Modulate Local Immune Response and Interference with Tuberculosis Diagnosis in Laboratory Animal Models.Veterinary sciences · 2020Article
- Article
- Characteristics and Epidemiological Investigation of Paratuberculosis in Dairy Cattle in Tai'an, China.BioMed research international · 2020Article
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The enteropathy called paratuberculosis (PTB), which mainly affects ruminants and has a worldwide distribution, is caused by Mycobacterium avium subsp. paratuberculosis. This disease significantly reduces the cost-effectiveness of ruminant farms, and therefore, reliable and rapid detection methods are needed to control the spread of the bacterium in livestock and in the environment. The aim of this study was to identify a specific and sensitive combination of DNA extraction and amplification to detect M. avium subsp. paratuberculosis in feces. Negative bovine fecal samples were inoculated with increasing concentrations of two different bacterial strains (field and reference) to compare the performance of four extraction and five amplification protocols. The best results were obtained using the JohnePrep and MagMax extraction kits combined with an in-house triplex real-time PCR designed to detect IS900, ISMap02 (an insertion sequence of M. avium subsp. paratuberculosis present in 6 copies per genome), and an internal amplification control DNA simultaneously. These combinations detected 10 M. avium subsp. paratuberculosis cells/g of spiked feces. The triplex PCR detected 1 fg of genomic DNA extracted from the reference strain K10. The performance of the robotized version of the MagMax extraction kit combined with the IS900 and ISMap02 PCR was further evaluated using 615 archival fecal samples from the first sampling of nine Friesian cattle herds included in a PTB control program and followed up for at least 4 years. The analysis of the results obtained in this survey demonstrated that the diagnostic method was highly specific and sensitive for the detection of M. avium subsp. paratuberculosis in fecal samples from cattle and a very valuable tool to be used in PTB control programs.
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