ArticleJournal of clinical microbiology2014
High-throughput direct fecal PCR assay for detection of Mycobacterium avium subsp. paratuberculosis in sheep and cattle.
Article in Journal of clinical microbiology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 42 papers.
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Who cites it
42 citing papers in PubMed, 93 citations in OpenAlex.
- Evaluation of the Johne's disease surveillance program in dairy herds in Hokkaido, Japan, using a stochastic individual-based simulation model.The Journal of veterinary medical science · 2026Article
- Systematic evaluation and optimization of TaqMan qPCR assays targeting FJournal of clinical microbiology · 2026Article
- Rapid and Simple Detection ofPathogens (Basel, Switzerland) · 2025Article
- Prevalence and Genotyping ofVeterinary sciences · 2025Article
- First serological diagnosis of Mycobacterium avium subsp. paratuberculosis infection in sheep in the state of Pernambuco, Brazil.Veterinary research communications · 2024Article
- Inter-laboratory ring trial to compare four quantitative polymerase chain reaction assays employed for detection ofMicrobiology spectrum · 2024Article
- Advances in understanding the genetic architecture of antibody response to paratuberculosis in sheep by heritability estimate and LDLA mapping analyses and investigation of candidate regions using sequence-based data.Genetics, selection, evolution : GSE · 2024Article
- Application of Bayesian modeling for diagnostic assays of Mycobacterium avium subsp. paratuberculosis in sheep and goats flocks.BMC veterinary research · 2022Article
- Correlates of vaccine protection againstFrontiers in veterinary science · 2022Article
- Early response of monocyte-derived macrophages from vaccinated and non-vaccinated goats against in vitro infection with Mycobacterium avium subsp. paratuberculosis.Veterinary research · 2021Article
- Article
- 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
- Disposable silicon-based all-in-one micro-qPCR for rapid on-site detection of pathogens.Nature communications · 2020Article
- A Novel Real-Time PCR-Based Screening Test with Pooled Fecal Samples for Bovine Johne's Disease.Journal of clinical microbiology · 2020Article
- Long non-coding RNA Gm15441 attenuates hepatic inflammasome activation in response to PPARA agonism and fasting.Nature communications · 2020Article
- Development of a qPCR for the detection and quantification ofJournal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc · 2020Article
- Control of Mycobacterium avium subsp. paratuberculosis infection on a New Zealand pastoral dairy farm.BMC veterinary research · 2019Article
- The humoral immune response is essential for successful vaccine protection against paratuberculosis in sheep.BMC veterinary research · 2019Article
- Application of Transcriptomics to Enhance Early Diagnostics of Mycobacterial Infections, with an Emphasis onVeterinary sciences · 2019Review
- Gene expression profiles during subclinical Mycobacterium avium subspecies paratuberculosis infection in sheep can predict disease outcome.Scientific reports · 2019Article
Corrections and comments
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Authors and funding
10 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Johne's disease (JD) is a chronic enteric disease caused by Mycobacterium avium subsp. paratuberculosis that affects ruminants. Transmission occurs by the fecal-oral route. A commonly used antemortem diagnostic test for the detection of M. avium subsp. paratuberculosis in feces is liquid culture; however, a major constraint is the 2- to 3-month incubation period needed for this method. Rapid methods for the detection of M. avium subsp. paratuberculosis based on PCR have been reported, but comprehensive validation data are lacking. We describe here a new test, the high-throughput-Johnes (HT-J), to detect M. avium subsp. paratuberculosis in feces. Its diagnostic accuracy was compared with that of liquid radiometric (Bactec) fecal culture using samples from cattle (1,330 samples from 23 herds) and sheep (596 samples from 16 flocks). The multistage protocol involves the recovery of M. avium subsp. paratuberculosis cells from a fecal suspension, cell rupture by bead beating, extraction of DNA using magnetic beads, and IS900 quantitative PCR. The limit of detection of the assay was 0.0005 pg, and the limit of quantification was 0.005 pg M. avium subsp. paratuberculosis genomic DNA. Only M. avium subsp. paratuberculosis was detected from a panel of 51 mycobacterial isolates, including 10 with IS900-like sequences. Of the 549 culture-negative fecal samples from unexposed herds and flocks, 99% were negative in the HT-J test, while 60% of the bovine- and 84% of the ovine-culture-positive samples were positive in the HT-J test. As similar total numbers of samples from M. avium subsp. paratuberculosis-exposed animals were positive in culture and HT-J tests in both species, and as the results of a McNemar's test were not significant, these methods probably have similar sensitivities, but the true diagnostic sensitivities of these tests are unknown. These validation data meet the consensus-based reporting standards for diagnostic test accuracy studies for paratuberculosis and the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines (S. A. Bustin et al., Clin. Chem. 55:611-622, 2009, doi:10.1373/clinchem.2008.112797). The HT-J assay has been approved for use in JD control programs in Australia and New Zealand.
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