ArticlePloS one2013
Enhanced detection of tuberculous mycobacteria in animal tissues using a semi-nested probe-based real-time PCR.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers, 1 of them a synthesis that pooled it.
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Who cites it
21 citing papers in PubMed, 1 synthesis or guideline pooled it, 46 citations in OpenAlex.
- Accuracy of Tests for Diagnosis of Animal Tuberculosis: Moving Away from the Golden Calf (and towards Bayesian Models).Transboundary and emerging diseases · 2023Pooled it
- Shaken, not stirred: magnetic bead DNA extraction as a rapid and effective method for the scaling up of bovine tuberculosis diagnosis.BMC veterinary research · 2024Article
- Insights into mycobacteriome composition in Mycobacterium bovis-infected African buffalo (Syncerus caffer) tissue samples.Scientific reports · 2024Article
- Droplet digital PCR as alternative to microbiological culture forFrontiers in cellular and infection microbiology · 2024Article
- Optimization of real-time PCR protocols from lymph node bovine tissue for direct detection ofMicrobiology spectrum · 2023Article
- Presence of Foodborne Bacteria in Wild Boar and Wild Boar Meat-A Literature Survey for the Period 2012-2022.Foods (Basel, Switzerland) · 2023Review
- Real-time PCR using atpE, conventional PCR targeting different regions of difference, and flow cytometry for confirmation of Mycobacterium bovis in buffaloes and cattle from the Delta area of Egypt.BMC microbiology · 2022Article
- Direct PCR on Tissue Samples To Detect Mycobacterium tuberculosis Complex: an Alternative to the Bacteriological Culture.Journal of clinical microbiology · 2021Article
- Real-Time PCR Validation forFrontiers in veterinary science · 2021Article
- Histopathological and microbiological study of porcine lymphadenitis: contributions to diagnosis and control of the disease.Porcine health management · 2020Article
- A molecular strategy to optimize bovine tuberculosis post-mortem diagnosis and the exposure to Mycobacterium tuberculosis variant bovis.Molecular biology reports · 2020Article
- The rapid detection and differentiation of Mycobacterium tuberculosis complex members from cattle and water buffaloes in the delta area of Egypt, using a combination of real-time and conventional PCR.Molecular biology reports · 2019Article
- Validation of a Real-Time PCR for the Detection ofFrontiers in veterinary science · 2019Article
- Continuous Excretion ofThe American journal of tropical medicine and hygiene · 2017Article
- Exposure of Threatened Accipitridae to Mycobacterium bovis Calls for Active Surveillance.EcoHealth · 2017Article
- First Detection of Borrelia burgdorferi sensu lato DNA in Serum of the Wild Boar (Sus scrofa) in Northern Portugal by Nested-PCR.EcoHealth · 2015Article
- Multi-Etiological Nature of Tuberculosis-Like Lesions in Condemned Pigs at the Slaughterhouse.PloS one · 2015Article
- Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2014Article
- Standing of nucleic acid testing strategies in veterinary diagnosis laboratories to uncover Mycobacterium tuberculosis complex members.Frontiers in molecular biosciences · 2014Review
- Direct detection and differentiation of pathogenic Leptospira species using a multi-gene targeted real time PCR approach.PloS one · 2014Article
Corrections and comments
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Authors and funding
9 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Bovine tuberculosis has been tackled for decades by costly eradication programs in most developed countries, involving the laboratory testing of tissue samples from allegedly infected animals for detection of Mycobacterium tuberculosis complex (MTC) members, namely Mycobacterium bovis. Definitive diagnosis is usually achieved by bacteriological culture, which may take up to 6-12 weeks, during which the suspect animal carcass and herd are under sanitary arrest. In this work, a user-friendly DNA extraction protocol adapted for tissues was coupled with an IS6110-targeted semi-nested duplex real-time PCR assay to enhance the direct detection of MTC bacteria in animal specimens, reducing the time to achieve a diagnosis and, thus, potentially limiting the herd restriction period. The duplex use of a novel β-actin gene targeted probe, with complementary targets in most mammals, allowed the assessment of amplification inhibitors in the tissue samples. The assay was evaluated with a group of 128 fresh tissue specimens collected from bovines, wild boars, deer and foxes. Mycobacterium bovis was cultured from 57 of these samples. Overall, the full test performance corresponds to a diagnostic sensitivity and specificity of 98.2% (CIP95% 89.4-99.9%) and 88.7% (CIP95% 78.5-94.7%), respectively. An observed kappa coefficient was estimated in 0.859 (CI P95% 0.771-0.948) for the overall agreement between the semi-nested PCR assay and the bacteriological culture. Considering only bovine samples (n = 69), the diagnostic sensitivity and specificity were estimated in 100% (CIP95% 84.0-100%) and 97.7% (CIP95% 86.2-99.9%), respectively. Eight negative culture samples exhibiting TB-like lesions were detected by the semi-nested real-time PCR, thus emphasizing the increased potential of this molecular approach to detect MTC-infected animal tissues. This novel IS6110-targeted assay allows the fast detection of tuberculous mycobacteria in animal specimens with very high sensitivity and specificity, being amenable and cost effective for use in the routine veterinary diagnostic laboratory with further automation possibilities.
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