Rapid culture-based methods for drug-resistance detection in Mycobacterium tuberculosis

Palomino, Juan Carlos; Martin, Anandi; Groll, Andréa von; Portaels, Françoise

doi:10.1016/j.mimet.2008.06.015

Cited by 41 publications

(39 citation statements)

References 61 publications

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“…Microscopic observation of broth cultures is a lowtechnology but rapid testing method for tuberculosis [13]. When coupled with drug sensitivity testing, this assay has been called microscopic observation of drug sensitivity (MODS).…”

Section: Isolation and Culturementioning

confidence: 99%

Cost-effectiveness in the diagnosis of tuberculosis: choices in developing countries

Molicotti

Bua

Zanetti

2014

J Infect Dev Ctries

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Tuberculosis remains one of the major causes of global death from a single infectious agent. This situation is worsened by the HIV/AIDS pandemic because one-third of HIV/AIDS patients are co-infected with Mycobacterium tuberculosis. Failure to control the spread of tuberculosis is largely due to our inability to detect and treat all infectious cases of pulmonary tuberculosis in a timely manner, allowing continued M. tuberculosis transmission within communities. Diagnosis of tuberculosis can be made using indirect and direct methods. The indirect tests, such as interferon-gamma release assays, provide a new diagnostic method for M. tuberculosis infection, but do not discriminate between infection and active disease. The most common direct method for diagnosing TB worldwide is sputum smear microscopy (developed more than 100 years ago), where bacteria are observed in sputum samples examined under a microscope. In countries with more developed laboratory capacities, cases of tuberculosis may also be diagnosed using culture methods (the current gold standard) or, increasingly, using rapid molecular tests. In this review, we discuss the traditional methods for the diagnosis of tuberculosis. We also discuss other inexpensive assays that can be used to detect the presence of M. tuberculosis.

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Section: Isolation and Culturementioning

confidence: 99%

Cost-effectiveness in the diagnosis of tuberculosis: choices in developing countries

Molicotti

Bua

Zanetti

2014

J Infect Dev Ctries

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“…ycobacterium tuberculosis grows very slowly, with a population doubling time of ϳ22 h. Consequently, detection of M. tuberculosis by outgrowth of CFU on solid culture medium requires weeks or months (17). Despite being labor-intensive and time-consuming, enumeration of CFU by plating sputum cultures on solid medium continues to be the "gold standard" for evaluating the effectiveness of antituberculosis chemotherapy (6,19).…”

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confidence: 99%

Malachite Green Interferes with Postantibiotic Recovery of Mycobacteria

Gelman

McKinney

Dhar

2012

Antimicrob Agents Chemother

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The genus Mycobacterium comprises slow-growing species with generation times ranging from hours to weeks. The protracted incubation time before colonies appear on solid culture medium can result in overgrowth by faster-growing microorganisms. To prevent contamination, the solid media used in laboratories and clinics for cultivation of mycobacteria contain the arylmethane compound malachite green, which has broad-spectrum antimicrobial activity. Malachite green has no impact on the plating efficiency of mycobacteria when cells are grown under normal conditions. However, we found that malachite green interfered with colony formation when bacteria were preexposed to antibiotics targeting cell wall biogenesis (isoniazid, ethionamide, ethambutol). This inhibitory effect of malachite green was not observed when bacteria were preexposed to antibiotics targeting cellular processes other than cell wall biogenesis (rifampin, moxifloxacin, streptomycin). Sputum specimens from tuberculosis patients are routinely evaluated on solid culture medium containing high concentrations of malachite green. This practice could lead to underestimation of bacterial loads and overestimation of chemotherapeutic efficacy. Mycobacterium tuberculosis grows very slowly, with a population doubling time of ϳ22 h. Consequently, detection of M. tuberculosis by outgrowth of CFU on solid culture medium requires weeks or months (17). Despite being labor-intensive and time-consuming, enumeration of CFU by plating sputum cultures on solid medium continues to be the "gold standard" for evaluating the effectiveness of antituberculosis chemotherapy (6,19). Contamination by faster-growing microorganisms is prevented by supplementing the medium with compounds that suppress the growth of common species in the human flora.In clinical practice, sputum samples from tuberculosis patients are routinely plated on Lowenstein-Jensen or Middlebrook 7H11 agar. These media contain high concentrations of malachite green, a diamino-triphenylmethane dye with broad-spectrum antimicrobial activity against Gram-positive and Gram-negative bacteria (1). The mechanistic basis of malachite green's antimicrobial activity is not understood, nor is it clear why mycobacteria can survive and replicate in the presence of high concentrations of this compound (1,12). This paper shows that malachite green interferes with the recovery of mycobacteria on solid culture medium following exposure to certain antibiotics. This inhibitory effect was specific to antibiotics that target cell wall biogenesis (isoniazid, ethionamide, ethambutol) and was not observed with antibiotics that target other cellular processes (rifampin, moxifloxacin, streptomycin). These observations may have implications for clinical practices because underestimation of bacterial loads in patients undergoing chemotherapy could result in overestimation of therapeutic efficacy. MATERIALS AND METHODSBacteria and culture conditions. Wild-type Mycobacterium smegmatis (strain mc 2 155) and M. tuberculosis (strain Erdman) were s...

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“…Unfortunately, this method often lacks sensitivity, as it is dependent on the organism burden in sputum samples [95]. Furthermore, classic culture-based methods to confirm the presence of M. tuberculosis using either liquid or solid culture media are cumbersome and lengthy and often performed only in specialty hospitals [92,[96][97][98]. This is particularly the case in countries with resource poor settings.…”

Section: Diagnosis and Treatment: Implications For Public Healthmentioning

confidence: 99%

“…Detection of TB and AST using culture-based methods can take 6 weeks or longer because of the slow growth of mycobacteria. In recent years, with the improvement of molecular diagnostics, several newer test methods for rapid detection and AST in TB have been developed and are commercially available [97,98]. With the advances in molecular technologies specifically during the past 10 years, several molecular test methods for the detection of M. tuberculosis and select resistance genes are now available.…”

Section: Diagnosis and Treatment: Implications For Public Healthmentioning

confidence: 99%

“…Various techniques are used for detection of M. tuberculosis: polymerase chain reaction (PCR), transcription-mediated amplification, strand displacement amplification, and loop-mediated isothermal amplification (LAMP). In various studies, summarized in a review by Parsons and colleagues, these tests have been shown a sensitivity of 86% -100% in smear-positive samples and 33% -93% in smear-negative samples [97]. Although most of these tests can be performed within 8 hours, it is important to recognize that initial steps of specimen (sputum) processing are still necessary.…”

Section: Diagnosis and Treatment: Implications For Public Healthmentioning

confidence: 99%

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Tuberculosis: From an Untreatable Disease in Antiquity to an Untreatable Disease in Modern Times?

Boire¹

2013

J Anc Dis Prev Rem

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The history of tuberculosis (TB) is intricately connected to the history of humanity. The disease is considered one of the oldest infectious diseases afflicting mankind. Its history is that of colorful, often vibrant descriptions and interpretations, in the attempt of human societies to demystify the origins, causality, and course of this grave and lethal disease, and in the ultimate pursuit of finding a cure. The discovery of the tubercle bacillus on March 24 th 1882, by Robert Koch, led to an unprecedented increase in international research efforts, ultimately resulting in the development of a vaccine and many potent antimicrobial agents and treatment regimens. However, the course of history is often not without some irony, commonly perceived as being unpredictable by those who find themselves immersed in history's path. In this sense, and despite the advances that were made in diagnostics and treatment during the past 70 years, TB continues to challenge mankind on numerous levels even today. The most recent emergence of multidrug-resistant and extensively-drug-resistant strains of Mycobacterium tuberculosis is only a small but significant reflection of the ongoing challenges in the attempt of eradicating this disease. Here we provide a review of the historic aspects of TB leading to a discussion of the current state of the approach to antituberculous treatment, including the aspects of microbiology, diagnostics, antimicrobial therapy, and public health.

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Rapid culture-based methods for drug-resistance detection in Mycobacterium tuberculosis

Cited by 41 publications

References 61 publications

Cost-effectiveness in the diagnosis of tuberculosis: choices in developing countries

Cost-effectiveness in the diagnosis of tuberculosis: choices in developing countries

Malachite Green Interferes with Postantibiotic Recovery of Mycobacteria

Tuberculosis: From an Untreatable Disease in Antiquity to an Untreatable Disease in Modern Times?

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