Novel Multiplex Real-Time PCR Diagnostic Assay for Identification and Differentiation of Mycobacterium tuberculosis, Mycobacterium canettii, and Mycobacterium tuberculosis Complex Strains

Autor: Majella Maher, Dick van Soolingen, Thomas Barry, Siobhan Dorai-Raj, Justin O'Grady, Kate Reddington
Přispěvatelé: College of Sciences, National University of Ireland Galway funded KR Postgraduate Scholarship
Rok vydání: 2011
Předmět:
DNA
Bacterial
Microbiology (medical)
Time Factors
Tuberculosis
medicine.drug_class
Molecular Sequence Data
Antibiotics
bovis bcg
comparative genomics
pinnipedii
Polymerase Chain Reaction
Sensitivity and Specificity
Microbiology
Mycobacterium
law.invention
members
Mycobacterium tuberculosis
Bacterial Proteins
Tuberculosis diagnosis
law
Mycobacterium tuberculosis complex (MTC)
medicine
Humans
Multiplex
discriminatory power
gene
reproducibility
Polymerase chain reaction
DNA Primers
Bacteriological Techniques
biology
Mycobacterium canettii
restriction analysis
Mycobacteriology and Aerobic Actinomycetes
Pathogenesis and modulation of inflammation Infection and autoimmunity [N4i 1]
Sequence Analysis
DNA
biology.organism_classification
medicine.disease
Virology
Mycobacterium tuberculosis complex
rapid identification
polymorphisms
Zdroj: Journal of Clinical Microbiology, 49, 2, pp. 651-7
Europe PubMed Central
Journal of Clinical Microbiology, 49, 651-7
ISSN: 1098-660X
0095-1137
DOI: 10.1128/jcm.01426-10
Popis: Tuberculosis (TB) in humans is caused by members of the Mycobacterium tuberculosis 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 M. tuberculosis and M. canettii . The lepA gene was targeted for the detection of members of the MTC, the wbbl1 gene was used for the differentiation of M. tuberculosis and M. canettii from the remainder of the complex, and a unique region of the M. canettii genome, a possible novel region of difference (RD), was targeted for the specific identification of M. canettii . 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 M. tuberculosis / M. canettii , the MTC, and M. canettii , 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 M. tuberculosis , M. canettii , and the other members of the MTC.
Databáze: OpenAIRE
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