Combined species identification, genotyping, and drug resistance detection of Mycobacterium tuberculosis cultures by MLPA on a bead-based array
| Autor: | Nino Tadumaze, Nino Bablishvili, Sarah Sengstake, Kiki Tuin, Anita C. Schürch, Elizabeta Bachiyska, Nadia Brankova, Viktoria Levterova, Christophe Sola, Dick van Soolingen, Maka Akhalaia, Frank Cobelens, Stefan Panaiotov, A. R. J. Schuitema, Anja van’t Hoog, Richard M. Anthony, Rina de Zwaan, Paul R. Klatser, Edgar Abadia, Rusudan Aspindzelashvili, Indra Bergval, Todor Kantardjiev |
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| Přispěvatelé: | Institut de génétique et microbiologie [Orsay] (IGM), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), KIT: Biomedical Research, Global Health, Infectious diseases |
| Rok vydání: | 2012 |
| Předmět: |
Bacterial Diseases
Applied Microbiology Extensively Drug-Resistant Tuberculosis Gene Identification and Analysis lcsh:Medicine Bovine Tuberculosis in Humans Poverty-related infectious diseases Infection and autoimmunity [N4i 3] Multiplex lcsh:Science Genetics 0303 health sciences Multidisciplinary biology Multi-Drug-Resistant Tuberculosis Microbial Mutation Nontuberculous Mycobacteria Mycobacterium Avium Complex Bacterial Pathogens 3. Good health Infectious Diseases [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology Mycobacterium tuberculosis complex Medicine Mycobacterium fortuitum Research Article Genotype Microbiology Polymorphism Single Nucleotide Mycobacterium Molecular Genetics Mycobacterium tuberculosis 03 medical and health sciences Genetic Mutation Microbial Control Drug Resistance Bacterial Multiplex polymerase chain reaction Tuberculosis [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Multiplex ligation-dependent probe amplification Biology Microbial Pathogens Genotyping 030304 developmental biology Mycobacterium kansasii 030306 microbiology lcsh:R Bacteriology biology.organism_classification lcsh:Q Multiplex Polymerase Chain Reaction |
| Zdroj: | PLoS ONE PLoS ONE, Public Library of Science, 2012, 7 (8), pp.e43240. ⟨10.1371/journal.pone.0043240⟩ PLoS ONE, Vol 7, Iss 8, p e43240 (2012) PLoS One, 7, 8 PLoS ONE, 7(8). Public Library of Science PLoS One, 7 |
| ISSN: | 1932-6203 |
| Popis: | Contains fulltext : 124255.pdf (Publisher’s version ) (Open Access) The population structure of Mycobacterium tuberculosis is typically clonal therefore genotypic lineages can be unequivocally identified by characteristic markers such as mutations or genomic deletions. In addition, drug resistance is mainly mediated by mutations. These issues make multiplexed detection of selected mutations potentially a very powerful tool to characterise Mycobacterium tuberculosis. We used Multiplex Ligation-dependent Probe Amplification (MLPA) to screen for dispersed mutations, which can be successfully applied to Mycobacterium tuberculosis as was previously shown. Here we selected 47 discriminative and informative markers and designed MLPA probes accordingly to allow analysis with a liquid bead array and robust reader (Luminex MAGPIX technology). To validate the bead-based MLPA, we screened a panel of 88 selected strains, previously characterised by other methods with the developed multiplex assay using automated positive and negative calling. In total 3059 characteristics were screened and 3034 (99.2%) were consistent with previous molecular characterizations, of which 2056 (67.2%) were directly supported by other molecular methods, and 978 (32.0%) were consistent with but not directly supported by previous molecular characterizations. Results directly conflicting or inconsistent with previous methods, were obtained for 25 (0.8%) of the characteristics tested. Here we report the validation of the bead-based MLPA and demonstrate its potential to simultaneously identify a range of drug resistance markers, discriminate the species within the Mycobacterium tuberculosis complex, determine the genetic lineage and detect and identify the clinically most relevant non-tuberculous mycobacterial species. The detection of multiple genetic markers in clinically derived Mycobacterium tuberculosis strains with a multiplex assay could reduce the number of TB-dedicated screening methods needed for full characterization. Additionally, as a proportion of the markers screened are specific to certain Mycobacterium tuberculosis lineages each profile can be checked for internal consistency. Strain characterization can allow selection of appropriate treatment and thereby improve treatment outcome and patient management. |
| Databáze: | OpenAIRE |
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