ArticleJournal of clinical microbiology2005
Development of a nested PCR method targeting a unique multicopy element, ISMap02, for detection of Mycobacterium avium subsp. paratuberculosis in fecal samples.
Article in Journal of clinical microbiology, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 33 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
33 citing papers in PubMed, 94 citations in OpenAlex.
- Evaluation of novel oral vaccine candidates and validation of a caprine model of Johne's disease.Frontiers in cellular and infection microbiology · 2014Trial
- East AsianJournal of veterinary science · 2026Article
- Neonatal Calf Serum MAP Antibody Titre as a Potential Marker of Early-Life MAP Exposure.Animals : an open access journal from MDPI · 2026Article
- Systematic evaluation and optimization of TaqMan qPCR assays targeting FJournal of clinical microbiology · 2026Article
- Development of visual dye-based loop-mediated isothermal amplification assays forJournal of veterinary science · 2025Article
- Inter-laboratory ring trial to compare four quantitative polymerase chain reaction assays employed for detection ofMicrobiology spectrum · 2024Article
- Comparison of 2 PCR assays on environmental samples cultured forJournal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc · 2024Article
- Bioprospective Role ofMolecules (Basel, Switzerland) · 2023Review
- Genomic epidemiology ofFrontiers in genetics · 2023Article
- Using Omics Approaches in the Discovery of Biomarkers for Early Diagnosis of Johne's Disease in Sheep and Goats.Animals : an open access journal from MDPI · 2021Review
- MAP, Johne's disease and the microbiome; current knowledge and future considerations.Animal microbiome · 2021Review
- Serological and Molecular Characterization ofAnimals : an open access journal from MDPI · 2021Article
- Alpha-2-Macroglobulin as a New Promising Biomarker Improving the Diagnostic Sensitivity of Bovine Paratuberculosis.Frontiers in veterinary science · 2021Article
- Detection ofFrontiers in veterinary science · 2021Article
- A pilot study using environmental screening to determine the prevalence ofIrish veterinary journal · 2020Article
- Isolation ofAnimals : an open access journal from MDPI · 2019Article
- ISMapVeterinary world · 2018Article
- Pathogenesis, Molecular Genetics, and Genomics ofFrontiers in veterinary science · 2017Review
- PCR-based detection of Mycobacterium avium subsp. paratuberculosis infection in cattle in South Korea using fecal samples.The Journal of veterinary medical science · 2016Article
- Evaluation of fecal culture and fecal RT-PCR to detect Mycobacterium avium ssp. paratuberculosis fecal shedding in dairy goats and dairy sheep using latent class Bayesian modeling.BMC veterinary research · 2016Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
This study describes the development of a nested PCR assay that uses a unique element (ISMap02) for Mycobacterium avium subsp. paratuberculosis that is present at six copies within the genome. In addition, the sensitivity of the assay with this element was compared to the sensitivity of detection of the IS900 element in both conventional and real-time PCR assays. The specificity of the ISMap02 element was evaluated by PCR of the DNA extracted from isolates of M. avium subsp. paratuberculosis and M. avium subsp. avium, as well as DNA from M. fortuitum, M. scofulaceum, M. phlei, M. smegmatis, and M. gordonae. Only M. avium subsp. paratuberculosis DNA was detectable after amplification with the ISMap02 primers. The sensitivity of detection for the ISMap02 element in either a conventional or a real-time PCR format was less than 100 fg DNA or 10(2) CFU/ml in serial titration curves with pure bacteria. These results were comparable to those obtained for the IS900 element. Experimental spiking of a negative fecal sample followed by M. avium subsp. paratuberculosis DNA extraction resulted in detection thresholds of 10(2) CFU/g for the IS900 element and 10(3) CFU/g for the ISMap02 element by using a real-time PCR format, but this sensitivity dropped 10-fold for both elements in a conventional PCR format. Analyses of fecal samples obtained from naturally infected animals demonstrated a sensitivity for the detection of M. avium subsp. paratuberculosis DNA by use of the ISMap02 element similar to that achieved by use of the IS900 element when it was used in a conventional PCR format. The real-time PCR format improved the levels of detection of both elements, but not to a significant degree. In conclusion, the ISMap02 element provides a very sensitive and specific alternative as a diagnostic reagent for use in PCR assays for the detection of paratuberculosis.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.