{"doi":"10.1128/jcm.01426-10","title":"Novel Multiplex Real-Time PCR Diagnostic Assay for Identification and Differentiation of Mycobacterium tuberculosis, Mycobacterium canettii, and Mycobacterium tuberculosis Complex Strains","abstract":"<jats:title>ABSTRACT</jats:title>\n          <jats:p>\n            Tuberculosis (TB) in humans is caused by members of the\n            <jats:named-content content-type=\"genus-species\">Mycobacterium tuberculosis</jats:named-content>\n            complex (MTC). Rapid detection of the MTC is necessary for the timely initiation of antibiotic treatment, while differentiation between members of the complex may be important to guide the appropriate antibiotic treatment and provide epidemiological information. In this study, a multiplex real-time PCR diagnostics assay using novel molecular targets was designed to identify the MTC while simultaneously differentiating between\n            <jats:named-content content-type=\"genus-species\">M. tuberculosis</jats:named-content>\n            and\n            <jats:named-content content-type=\"genus-species\">M. canettii</jats:named-content>\n            . The\n            <jats:italic>lepA</jats:italic>\n            gene was targeted for the detection of members of the MTC, the\n            <jats:italic>wbbl1</jats:italic>\n            gene was used for the differentiation of\n            <jats:named-content content-type=\"genus-species\">M. tuberculosis</jats:named-content>\n            and\n            <jats:named-content content-type=\"genus-species\">M. canettii</jats:named-content>\n            from the remainder of the complex, and a unique region of the\n            <jats:named-content content-type=\"genus-species\">M. canettii</jats:named-content>\n            genome, a possible novel region of difference (RD), was targeted for the specific identification of\n            <jats:named-content content-type=\"genus-species\">M. canettii</jats:named-content>\n            . The multiplex real-time PCR assay was tested using 125 bacterial strains (64 MTC isolates, 44 nontuberculosis mycobacteria [NTM], and 17 other bacteria). The assay was determined to be 100% specific for the mycobacteria tested. Limits of detection of 2.2, 2.17, and 0.73 cell equivalents were determined for\n            <jats:named-content content-type=\"genus-species\">M. tuberculosis</jats:named-content>\n            /\n            <jats:named-content content-type=\"genus-species\">M. canettii</jats:named-content>\n            , the MTC, and\n            <jats:named-content content-type=\"genus-species\">M. canettii</jats:named-content>\n            , respectively, using probit regression analysis. Further validation of this diagnostics assay, using clinical samples, should demonstrate its potential for the rapid, accurate, and sensitive diagnosis of TB caused by\n            <jats:named-content content-type=\"genus-species\">M. tuberculosis</jats:named-content>\n            ,\n            <jats:named-content content-type=\"genus-species\">M. canettii</jats:named-content>\n            , and the other members of the MTC.\n          </jats:p>","journal":"Journal of Clinical Microbiology","year":2011,"id":55979,"datarank":1.3645814351372927,"base_score":3.4011973816621555,"endowment":3.4011973816621555,"self_citation_contribution":0.5101796072493234,"citation_network_contribution":0.8544018278879691,"self_endowment_contribution":0.5101796072493234,"citer_contribution":0.8544018278879691,"corpus_percentile":null,"corpus_rank":null,"citation_count":29,"citer_count":25,"citers_with_citation_signal":18,"citers_with_endowment":18,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":290678,"name":"Justin O'Grady","orcid":null,"position":1,"is_corresponding":false},{"id":290679,"name":"Siobhan Dorai-Raj","orcid":null,"position":2,"is_corresponding":false},{"id":290680,"name":"Majella Maher","orcid":null,"position":3,"is_corresponding":false},{"id":290681,"name":"Dick van Soolingen","orcid":null,"position":4,"is_corresponding":false},{"id":290682,"name":"Thomas Barry","orcid":null,"position":5,"is_corresponding":false},{"id":290677,"name":"Kate Reddington","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Novel Multiplex Real-Time PCR Diagnostic Assay for Identification and Differentiation of Mycobacterium tuberculosis, Mycobacterium canettii, and Mycobacterium tuberculosis Complex Strains","abstract":"<jats:title>ABSTRACT</jats:title>\n          <jats:p>\n            Tuberculosis (TB) in humans is caused by members of the\n            <jats:named-content content-type=\"genus-species\">Mycobacterium tuberculosis</jats:named-content>\n            complex (MTC). Rapid detection of the MTC is necessary for the timely initiation of antibiotic treatment, while differentiation between members of the complex may be important to guide the appropriate antibiotic treatment and provide epidemiological information. In this study, a multiplex real-time PCR diagnostics assay using novel molecular targets was designed to identify the MTC while simultaneously differentiating between\n            <jats:named-content content-type=\"genus-species\">M. tuberculosis</jats:named-content>\n            and\n            <jats:named-content content-type=\"genus-species\">M. canettii</jats:named-content>\n            . The\n            <jats:italic>lepA</jats:italic>\n            gene was targeted for the detection of members of the MTC, the\n            <jats:italic>wbbl1</jats:italic>\n            gene was used for the differentiation of\n            <jats:named-content content-type=\"genus-species\">M. tuberculosis</jats:named-content>\n            and\n            <jats:named-content content-type=\"genus-species\">M. canettii</jats:named-content>\n            from the remainder of the complex, and a unique region of the\n            <jats:named-content content-type=\"genus-species\">M. canettii</jats:named-content>\n            genome, a possible novel region of difference (RD), was targeted for the specific identification of\n            <jats:named-content content-type=\"genus-species\">M. canettii</jats:named-content>\n            . The multiplex real-time PCR assay was tested using 125 bacterial strains (64 MTC isolates, 44 nontuberculosis mycobacteria [NTM], and 17 other bacteria). The assay was determined to be 100% specific for the mycobacteria tested. Limits of detection of 2.2, 2.17, and 0.73 cell equivalents were determined for\n            <jats:named-content content-type=\"genus-species\">M. tuberculosis</jats:named-content>\n            /\n            <jats:named-content content-type=\"genus-species\">M. canettii</jats:named-content>\n            , the MTC, and\n            <jats:named-content content-type=\"genus-species\">M. canettii</jats:named-content>\n            , respectively, using probit regression analysis. Further validation of this diagnostics assay, using clinical samples, should demonstrate its potential for the rapid, accurate, and sensitive diagnosis of TB caused by\n            <jats:named-content content-type=\"genus-species\">M. tuberculosis</jats:named-content>\n            ,\n            <jats:named-content content-type=\"genus-species\">M. canettii</jats:named-content>\n            , and the other members of the MTC.\n          </jats:p>","is_dataset_classified":null,"base_score":3.4011973816621555,"endowment":3.4011973816621555,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"21123525","pmcid":"PMC3043491","openalex_id":"https://openalex.org/W2149405576","authors":[],"funders":[{"funder_name":"Wellcome Trust","grant_id":"unidentified","title":"unidentified"}],"total_grants":1,"fwci":1.8047,"citation_percentile":0.85326814,"influential_citations":1,"citation_trend":[{"year":2012,"count":5},{"year":2013,"count":2},{"year":2014,"count":3},{"year":2015,"count":4},{"year":2016,"count":1},{"year":2017,"count":2},{"year":2018,"count":3},{"year":2019,"count":1},{"year":2022,"count":1},{"year":2024,"count":2},{"year":2026,"count":1}],"oa_status":"green","license":"ASM Journals Non-Commercial TDM","oa_locations":[{"url":"https://ueaeprints.uea.ac.uk/id/eprint/41293/1/Reddington_et_al_multiplex_1_2011.pdf","host_type":"repository"},{"url":"https://europepmc.org/articles/pmc3043491?pdf=render","host_type":"GREEN"},{"url":"https://ueaeprints.uea.ac.uk/id/eprint/41293/1/Reddington_et_al_multiplex_1_2011.pdf","host_type":"repository"},{"url":"https://journals.asm.org/doi/pdf/10.1128/JCM.01426-10","host_type":"publisher"},{"url":"https://doi.org/10.1128/jcm.01426-10","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/21123525","host_type":"repository"},{"url":"http://hdl.handle.net/10379/2563","host_type":"repository"},{"url":"http://hdl.handle.net/10379/13611","host_type":"repository"},{"url":"http://discovery.ucl.ac.uk/403452/","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/3043491","host_type":"repository"},{"url":"http://hdl.handle.net/2066/97169","host_type":"repository"},{"url":"https://rivm.openrepository.com/handle/10029/645531","host_type":"repository"},{"url":"https://jcm.asm.org/content/49/2/651.full.pdf","host_type":""},{"url":"https://dx.doi.org/10.13025/11103","host_type":""},{"url":"https://dx.doi.org/10.13025/15522","host_type":""},{"url":"https://doi.org/10.1128/JCM.01426-10","host_type":""},{"url":"https://dx.doi.org/10.1128/jcm.01426-10","host_type":""},{"url":"https://doi.org/https://doi.org/10.1128/JCM.01426-10","host_type":""}],"fields_of_study":["Mycobacterium research and diagnosis","Tuberculosis Research and Epidemiology","Infectious Diseases and Tuberculosis","Medicine","Biology","0301 basic medicine","03 medical and health sciences","Bacterial Proteins","Bacteriological Techniques","DNA Primers","DNA, Bacterial","Humans","Molecular Sequence Data","Mycobacterium","Polymerase Chain Reaction","Sensitivity and Specificity","Sequence Analysis, DNA","Time Factors","Tuberculosis"],"mesh_terms":["Bacterial Proteins","Bacteriological Techniques","DNA, Bacterial","Humans","Molecular Sequence Data","Mycobacterium","Sensitivity and Specificity","Time Factors","Tuberculosis","Polymerase Chain Reaction","Sequence Analysis, DNA","DNA Primers"],"keywords":["Mycobacterium tuberculosis","Mycobacterium tuberculosis complex","Tuberculosis","Multiplex","Multiplex polymerase chain reaction","Biology","Mycobacterium","Microbiology","Nontuberculous mycobacteria","Virology","Polymerase chain reaction","Medicine","Gene","Genetics","Pathology","DNA, Bacterial","Time Factors","Molecular Sequence Data","610","bovis bcg","comparative genomics","pinnipedii","Sensitivity and Specificity","members","Bacterial Proteins","Mycobacterium tuberculosis complex (MTC)","Humans","discriminatory power","reproducibility","DNA Primers","Bacteriological Techniques","Mycobacterium canettii","restriction analysis","Sequence Analysis, DNA","540","FOS: Biological sciences","rapid identification","polymorphisms","N4i 1: Pathogenesis and modulation of inflammation NCMLS 1: Infection and autoimmunity"],"sdg_mappings":[{"sdg_number":3,"sdg_label":"3. 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