Rapid and specific detection of the Mycobacterium tuberculosis complex using fluorogenic probes andreal-time PCR

Kraus, Günter; Cleary, Timothy J.; Miller, Nancimae; Seivright, Rachel; Young, Andrea K.; Spruill, Gina; Hnatyszyn, H. James

doi:10.1006/mcpr.2001.0385

Cited by 50 publications

(46 citation statements)

References 27 publications

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“…This method could be used in combination with any of the previously described methods to identify M. tuberculosis (12,13,15), and the simultaneous identification and detection of rifampin and isoniazid resistance in AR-positive sputa is possible.…”

Section: Discussionmentioning

confidence: 99%

Direct Detection of Rifampin- and Isoniazid-Resistant Mycobacterium tuberculosis in Auramine-Rhodamine-Positive Sputum Specimens by Real-Time PCR

et al. 2004

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Our objective was to evaluate the feasibility of a molecular assay based on a real-time PCR technique, carried out with a LightCycler instrument (Roche Biochemicals), to identify Mycobacterium tuberculosis bacilli and to detect rifampin and isoniazid resistance in DNA extracts from sputum samples. We studied three genes: rpoB, which is associated with rifampin resistance, and katG and inhA, which are associated with isoniazid resistance. A total of 205 sputum samples collected from 108 patients diagnosed with pulmonary tuberculosis with positive auramine-rhodamine-staining (AR) sputum samples, were tested. The sensitivities of the LightCycler PCR assay for the positive AR specimens was 97.5% (200 of 205) for rpoB and inhA genes and 96.5% (198 of 205) for the katG gene. For the total number of patients tested, the sensitivity was 100% (108 of 108 patients) for rifampin, whereas the sensitivity was 98.1% (106 of 108 patients) for isoniazid. Full agreement was found with the Bactec MGIT 960 method and the genotype inferred from the LightCycler data for rifampin. The phenotypic method for isoniazid reported 13 resistant strains (>0.1 g/ml). In seven (53.8%) strains there was a concordance between both methods, but we found that six (46.2%) strains reported as resistant by the phenotypic method were determined to be susceptible by real-time PCR. For the 75 strains reported as susceptible by the phenotypic method, the concordance with the LightCycler data was 100%. Our results demonstrate that rifampin-resistant M. tuberculosis could be detected in DNA extracted from auraminerhodamine-positive sputum samples in a single-tube assay that took less than 3 h to perform for a collection of auramine-rhodamine-positive specimens obtained from patients with culture-documented pulmonary tuberculosis. Similarly, this occurs in half of the isoniazid-resistant M. tuberculosis DNA extracted from auramine-rhodamine-positive specimens.

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Section: Discussionmentioning

confidence: 99%

Direct Detection of Rifampin- and Isoniazid-Resistant Mycobacterium tuberculosis in Auramine-Rhodamine-Positive Sputum Specimens by Real-Time PCR

et al. 2004

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“…Real-time detection technology has made it possible to establish non-commercial, probe-based PCR systems that provide stable operation, low contamination risk and automated interpretation of results (12). Detection of M. tuberculosis complex (MTBC) in the extrapulmonary mode of the disease through these advanced diagnostic tools has not received the attention it is due.…”

Section: Introductionmentioning

confidence: 99%

Molecular detection of Mycobacterium tuberculosis in formalin-fixed, paraffin-embedded tissues and biopsies of gastrointestinal specimens using real-time polymerase chain reaction system

Mishra

Gorantla²,

Bhargava³

et al. 2010

Turk J Gastroenterol

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“…This system has been used successfully for the rapid detection or identification of a variety of microorganisms (8-10, 15, 16, 19-21, 23). Real-time PCR with fluorogenic probes has also been shown to be useful for the detection of mycobacteria, including M. tuberculosis (12,13,17). An added feature of the LightCycler real-time PCR system is the postamplification melting point analysis.…”

mentioning

confidence: 99%

Detection and Differentiation of Mycobacterium tuberculosis and Nontuberculous Mycobacterial Isolates by Real-Time PCR

et al. 2003

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Rapid and specific detection of the Mycobacterium tuberculosis complex using fluorogenic probes andreal-time PCR

Cited by 50 publications

References 27 publications

Direct Detection of Rifampin- and Isoniazid-Resistant Mycobacterium tuberculosis in Auramine-Rhodamine-Positive Sputum Specimens by Real-Time PCR

Direct Detection of Rifampin- and Isoniazid-Resistant Mycobacterium tuberculosis in Auramine-Rhodamine-Positive Sputum Specimens by Real-Time PCR

Molecular detection of Mycobacterium tuberculosis in formalin-fixed, paraffin-embedded tissues and biopsies of gastrointestinal specimens using real-time polymerase chain reaction system

Detection and Differentiation of Mycobacterium tuberculosis and Nontuberculous Mycobacterial Isolates by Real-Time PCR

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