A novel multiplex real-time PCR for the identification of mycobacteria associated with zoonotic tuberculosis
| Autor: | Dick van Soolingen, Siobhan Dorai-Raj, Justin O'Grady, Thomas Barry, Kate Reddington, Stefan Niemann |
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| Přispěvatelé: | College of Sciences, National University of Ireland Galway funded KR Postgraduate Scholarship |
| Rok vydání: | 2011 |
| Předmět: |
Bacterial Diseases
nontuberculous mycobacteria Bovine Tuberculosis in Humans comparative genomics law.invention law Zoonoses Mycobacterium tuberculosis complex (MTC) samples Multiplex Bovine Tuberculosis discriminatory power humans Polymerase chain reaction 0303 health sciences Mycobacterium bovis Multidisciplinary biology Zoonotic Diseases Bacterial Pathogens 3. Good health Infectious Diseases Veterinary Diseases Mycobacterium tuberculosis complex Medical Microbiology Medicine Research Article Neglected Tropical Diseases DNA Bacterial Tuberculosis Science Zoonotic Tuberculosis Real-Time Polymerase Chain Reaction Sensitivity and Specificity Microbiology Mycobacterium Mycobacterium tuberculosis bovis subsp caprae 03 medical and health sciences Species Specificity Tuberculosis diagnosis medicine Animals Biology Microbial Pathogens DNA Primers 030304 developmental biology 030306 microbiology Reproducibility of Results Pathogenesis and modulation of inflammation Infection and autoimmunity [N4i 1] Sequence Analysis DNA medicine.disease biology.organism_classification Virology infection clinical specimens Emerging Infectious Diseases Cattle Veterinary Science rapid identification Tuberculosis Bovine complex strains |
| Zdroj: | PLoS ONE, Vol 6, Iss 8, p e23481 (2011) PLoS ONE PLoS One, 6 Europe PubMed Central PLoS One, 6, 8 |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0023481 |
| Popis: | Contains fulltext : 95584.pdf (Publisher’s version ) (Open Access) BACKGROUND: Tuberculosis (TB) is the leading cause of death worldwide from a single infectious agent. An ability to detect the Mycobacterium tuberculosis complex (MTC) in clinical material while simultaneously differentiating its members is considered important. This allows for the gathering of epidemiological information pertaining to the prevalence, transmission and geographical distribution of the MTC, including those MTC members associated with zoonotic TB infection in humans. Also differentiating between members of the MTC provides the clinician with inherent MTC specific drug susceptibility profiles to guide appropriate chemotherapy. METHODOLOGY/PRINCIPAL FINDINGS: The aim of this study was to develop a multiplex real-time PCR assay using novel molecular targets to identify and differentiate between the phylogenetically closely related M. bovis, M. bovis BCG and M. caprae. The lpqT gene was explored for the collective identification of M. bovis, M. bovis BCG and M. caprae, the lepA gene was targeted for the specific identification of M. caprae and a Region of Difference 1 (RD1) assay was incorporated in the test to differentiate M. bovis BCG. The multiplex real-time PCR assay was evaluated on 133 bacterial strains and was determined to be 100% specific for the members of the MTC targeted. CONCLUSIONS/SIGNIFICANCE: The multiplex real-time PCR assay developed in this study is the first assay described for the identification and simultaneous differentiation of M. bovis, M. bovis BCG and M. caprae in one internally controlled reaction. Future validation of this multiplex assay should demonstrate its potential in the rapid and accurate diagnosis of TB caused by these three mycobacteria. Furthermore, the developed assay may be used in conjunction with a recently described multiplex real-time PCR assay for identification of the MTC and simultaneous differentiation of M. tuberculosis, M. canettii resulting in an ability to differentiate five of the eight members of the MTC. |
| Databáze: | OpenAIRE |
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