Detection of Rifampin Resistance in
            <i>Mycobacterium tuberculosis</i>
            in a Single Tube with Molecular Beacons

El-Hajj, Hiyam; Marras, Salvatore A. E.; Tyagi, Sanjay; Kramer, Fred Russell; Alland, David

doi:10.1128/jcm.39.11.4131-4137.2001

Cited by 158 publications

(123 citation statements)

References 28 publications

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“…It uses a hemi-nested real-time PCR assay to amplify an MTB-specific sequence of the rpoB gene, which is probed with molecular beacons for mutations in the rifampin resistance-determining region [57]. This test can yield results in 2 hours.…”

Section: Cohen Et Al Tuberculous Pleural Effusionmentioning

confidence: 99%

Tuberculous Pleural Effusion

Cohen

Light

2015

Tur Toraks Der

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When a patient presents with new pleural effusion, the diagnosis of tuberculous (TB) pleuritis should be considered. The patient is at risk for developing pulmonary or extrapulmonary TB if the diagnosis is not made. Between 3% and 25% of patients with TB will have TB pleuritis. The incidence of TB pleuritis is higher in patients who are human immunodeficiency virus (HIV)-positive. Pleural fluid is an exudate that usually has a predominance of lymphocytes. The easiest way to diagnose TB pleuritis in a patient with lymphocytic pleural effusion is to demonstrate a pleural fluid adenosine deaminase level above 40 IU/L. The treatment for TB pleuritis is the same as that for pulmonary TB. Tuberculous empyema is a rare occurrence, and the treatment is difficult. KEY WORDS:Tuberculosis, pleural effusions, adenosine deaminase, gamma interferon, pleural biopsy, empyema INTRODUCTION Tuberculosis (TB) remains a major public health problem in developing countries. Most patients with TB have pulmonary TB; however, 25% of adults can present with extrapulmonary TB, which mainly involves the lymph nodes and pleura but can affect any organ system, including the central nervous system (CNS), cardiovascular system, and gastrointestinal (GI) tract [1]. The most common form of extrapulmonary TB is infection of the pleura. In a patient who presents with undiagnosed pleural effusion, the diagnosis of TB pleuritis should be considered. Pleural effusion, as an isolated manifestation of TB, may be selflimited and of little immediate concern, but untreated, it can lead to serious disease many years later. Pathogenesis and Pathologic FeaturesTuberculous pleural effusion may occur in the absence of radiologically apparent TB [2]. The effusion may result as a sequela to a primary infection from 6-12 weeks earlier, or it may represent the reactivation of TB [2]. In developed countries, TB pleural effusions present more often in older patients. The median age in one North American study was 56, suggesting reactivation of the disease [3]. However, a study from 2004 in San Francisco showed that pleural TB cases were 2 times more likely to be clustered (when assessed by genotyping of the mycobacterial organisms) than pulmonary TB and 3 times more likely to be clustered than nonrespiratory TB cases [4]. The observations made in the San Francisco study suggest that the majority of patients has a postprimary infection [4]. This was not confirmed in studies performed in both Houston [5] and Baltimore [6] or in a study in sub-Saharan Africa, in which the prevalence of pleural TB was 63.2% in patients with a primary TB infection [7].The pathogenesis of TB pleural effusion is thought to be related to the rupture of a subpleural caseous focus in the lung into the pleural space [8]. The basis for this was the observation that a caseous TB focus could be demonstrated in the lung, contiguous with the diseased pleural, in 12 to 15 patients with TB pleuritis [9]. The 3 other patients in this study had parenchymal disease but did not have caseous foci adjacent...

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Section: Cohen Et Al Tuberculous Pleural Effusionmentioning

confidence: 99%

Tuberculous Pleural Effusion

Cohen

Light

2015

Tur Toraks Der

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“…For RT-PCR, 10 l of 10 4 cfu/ml M. tuberculosis (Erdman) were mixed with either 13 g of TDMH or an equivalent volume of storage buffer in PBSTA, incubated for 30 min at 37°C, and then heat-inactivated at 80°C for 20 min. After centrifugation at 14,000 rpm for 30 s, 1 l of the supernatant from the treated mixture was directly added as a template to the molecular beacon-based RT-PCR described previously (30), with 500 nM each of the M. tuberculosis 16 S rRNA-specific forward and reverse primers, 200 nM molecular beacon, and 5 l of 2ϫ RT-PCR master mix (Applied Biosystems). The amplification conditions were as follows: 95°C for 10 min, followed by 40 cycles of 95°C for 30 s, 58°C for 60 s, and 72°C for 30 s in a StepOnePlus RT-PCR System (Applied Biosystems).…”

Section: In Vitro Catalytic Activity Of Tdmh-[mentioning

confidence: 99%

Exposure to a Cutinase-like Serine Esterase Triggers Rapid Lysis of Multiple Mycobacterial Species

Yang

Bhatti

et al. 2013

Journal of Biological Chemistry

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Background: Mycobacteria possess a unique envelope with poorly understood architectural organization of constituent molecules. Results: Exposure to an esterase that targets trehalose mycolates triggers rapid lysis of mycobacteria. Conclusion: Trehalose mycolates are on the exposed surface of mycobacterial envelope with an indispensable structural role. Significance: The study identifies a novel enzyme-based lysis of mycobacteria, with a potential for new anti-tuberculosis applications.

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“…PCR-based methods with readouts including line probe assays, analysis with DNA microarrays and real-time PCR have been the focus of much recent effort in developing molecular DST methods to determine resistance to RIF and INH (Cirillo et al, 2004;Edwards et al, 2001;El-Hajj et al, 2001;Herrera-Leó n et al, 2005;van Doorn et al, 2003). Among the most successful of these molecular diagnostic methods is the GenoType MTBDRplus assay (Hain LifeScience), which is based on reverse hybridization of amplicons containing rpoB, katG and inhA from test strains to probes carrying specific mutations in these genes.…”

Section: Introductionmentioning

confidence: 99%

Improved rapid molecular diagnosis of multidrug-resistant tuberculosis using a new reverse hybridization assay, REBA MTB-MDR

Bang

Park

Hwang

et al. 2011

Journal of Medical Microbiology

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Rapid diagnosis of multidrug-resistant tuberculosis (MDR-TB) is essential for the prompt initiation of effective second-line therapy to improve treatment outcome and limit transmission of this obstinate disease. A variety of molecular methods that enable the rapid detection of mutations implicated in MDR-TB have been developed. The sensitivity of the methods is dependent, in principle, on the repertoire of mutations being detected, which is typically limited to mutations in the genes rpoB, katG and the promoter region of inhA. In this study, a new reverse hybridization assay, REBA MTB-MDR (M&D), that probes mutations in the oxyR-ahpC intergenic region, in addition to those in rpoB, katG and the inhA promoter region, was evaluated. A set of 240 Mycobacterium tuberculosis clinical isolates from patients receiving retreatment regimens was subjected to conventional phenotypic drug-susceptibility testing (DST) and the REBA MTB-MDR assay. The nucleotide sequences of the loci known to be involved in drug resistance were determined for comparison. In brief, the results showed that the REBA MTB-MDR assay efficiently recognized nucleotide changes in the oxyR-ahpC intergenic region as well as those in rpoB, katG and the inhA promoter region with higher sensitivity, resulting in an 81.0 % detection rate for isoniazid resistance. Inclusion of the oxyR-ahpC intergenic region in the REBA MTB-MDR assay improved the overall sensitivity of molecular DST for MDR-TB from 73.1 to 79.9 %.

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Detection of Rifampin Resistance in Mycobacterium tuberculosis in a Single Tube with Molecular Beacons

Abstract: Current clinical assays for determining antibiotic susceptibility in

Cited by 158 publications

References 28 publications

Tuberculous Pleural Effusion

Tuberculous Pleural Effusion

Exposure to a Cutinase-like Serine Esterase Triggers Rapid Lysis of Multiple Mycobacterial Species

Improved rapid molecular diagnosis of multidrug-resistant tuberculosis using a new reverse hybridization assay, REBA MTB-MDR

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