Novel Multiplex Real-Time PCR Diagnostic Assay for Identification and Differentiation of Mycobacterium tuberculosis, Mycobacterium canettii, and Mycobacterium tuberculosis Complex Strains
van Soolingen, Dick
MetadataShow full item record
This item's downloads: 1366 (view details)
Reddington, K., J. O'Grady, S. Dorai-Raj, M. Maher, D. van Soolingen, and T. Barry. 2011. Novel Multiplex Real-Time PCR Diagnostic Assay for Identification and Differentiation of Mycobacterium tuberculosis, Mycobacterium canettii, and Mycobacterium tuberculosis Complex Strains. J. Clin. Microbiol. 49:651-657
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.
This item is available under the Attribution-NonCommercial-NoDerivs 3.0 Ireland. No item may be reproduced for commercial purposes. Please refer to the publisher's URL where this is made available, or to notes contained in the item itself. Other terms may apply.
The following license files are associated with this item:
Showing items related by title, author, creator and subject.
Ryan, EG; Dwyer, PJ; Connolly, DJ; Fagan, J; Costello, E; More, SJ (Springer Nature, 2008-01-01)This case report describes tuberculosis (TB) due to infection with Mycobacterium bovis (M. bovis) in alpaca (Lama pacos) on a farm in Ireland. Two severely debilitated alpaca were presented to the University Veterinary ...
Tuberculosis in alpaca (lama pacos) on a farm in ireland. 2. results of an epidemiological investigation Connolly, DJ; Dwyer, PJ; Fagan, J; Hayes, M; Ryan, EG; Costello, E; Kilroy, A; More, SJ (Springer Nature, 2008-01-01)Tuberculosis (TB), due to infection with Mycobacterium bovis was diagnosed in a flock of alpaca in Ireland in 2004. An epidemiological investigation was conducted to identify the risk of TB for farmed alpaca where TB is ...
A Novel Multiplex Real-Time PCR for the Identification of Mycobacteria Associated with Zoonotic Tuberculosis Reddington, Kate; O'Grady, Justin; Dorai-Raj, Siobhan; Niemann, Stefan; van Soolingen, Dick; Barry, Thomas (PLoS ONE, 2011-08-09)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 ...