The thin-layer agar method for direct phenotypic detection of multi- and extensively drug-resistant tuberculosis

Ardizzoni, Elisa; Mulders, Wim; Kotrikadze, Tinatin; Aspindzelashvili, Rusudan; Goginashvili, Leila; Pangtey, H.; Varaine, Francis; Bastard, Mathieu; Rigouts, Leen; Jong, Bouke C. de

doi:10.5588/ijtld.15.0136

Cited by 13 publications

(26 citation statements)

References 10 publications

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“…The total agreement between the Colour Test and the MGIT 960 system was excellent with over 95% for all the anti-TB drugs tested (Fig 4). Furthermore, the specificities found in this study were > 97% for all technicians, which is in good correlation with previous studies using similar methods [23,24,28,36]. The sensitivities varied between the study laboratories being slightly higher in the Lab B for the detection of INH and RIF resistance and higher in the Lab A with 100% for the detection of LVX resistance.…”

Section: Resultssupporting

confidence: 90%

“…for isoniazid (INH), rifampicin (RIF) and ciprofloxacin (CFX) [23]. In former studies, where the performance of CT was evaluated, the media was prepared in an expert laboratory elsewhere and transported into the study site or the specimens from the study site were transported into an expert laboratory for testing [24,25]. Hence, the preparations of the media represent a constraint of the CT method.…”

Section: Introductionmentioning

confidence: 99%

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Successful introduction of the Colour Test into inexperienced settings

Klaos

Kummik

et al. 2020

Preprint

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Implementation of non-commercial in-house methods into routine clinical diagnostics becomes more challenging, because these methods are not internationally standardized, most of the research in that field is underfunded and recommendations for their use is lacking. We conducted a study, where all the technicians were previously unfamiliar to the Colour Test (CT), a colorimetric redox indicator and thin-layer agar based Mycobacterium tuberculosis complex diagnosis and direct drug susceptibility testing (DST) method, and tested whether the performance of this in-house method is dependent on experience of the laboratory personnel.After a two-day hands-on training, six panels of 150 M. tuberculosis isolates were cultured onto CT plates prepared in-house by six technicians in two laboratories. Finally, triplicate readings of 900 CT plates resulted 18 DST patterns for each of the initial isolates. The results were compared to each other and the gold standard of BACTEC MGIT 960 DST results.The median time to produce M. tuberculosis CT DST results for three antituberculosis drugs was 13 days. The overall ability to correctly define phenotypic resistance ranged from 94.7% for levofloxacin to 95.8% and 97.3% for isoniazid and rifampicin, respectively. The test specificities were even higher exceeding 97% for all three drugs tested. Interobserver agreement reached 100% in one of the laboratories and exceeded 97% for levofloxacin and 99% for isoniazid and rifampicin in the second laboratory.The implementation of the CT into a new laboratory was straightforward with only minimal guidance. This study proves that the CT is highly reproducible and easily interpreted by previously inexperienced personnel.

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Section: Resultssupporting

confidence: 90%

Section: Introductionmentioning

confidence: 99%

Successful introduction of the Colour Test into inexperienced settings

Klaos

Kummik

et al. 2020

Preprint

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“…These methods are costly, require infrastructure, and can test susceptibility of limited number of anti-TB drugs; so molecular methods cannot eliminate requirement of culture-based methods. 4,9,[12][13] Therefore, there is an urgent need for the development of a simple, rapid but cost effective technique for the detection of MDR and XDR-TB in developing countries. 6,11,14 To reduce the time of diagnosis and DST, several non-commercial methods such as calorimetric redox indicator (CRI) method, microscopic observation drug susceptibility (MODS) assay, nitrate reductase assay (NRA) and thin-layer agar (TLA) have been developed.…”

Section: Original Articlementioning

confidence: 99%

“…It has demonstrated good performance for INH and RIF. [5][6]10,12,14 The objective of this study was to evaluate TLA method for determing sensitivity of second-line drugs OFX and KM in AFB smear positive clinical specimens and compare it with the MGIT 960 system.…”

Section: Original Articlementioning

confidence: 99%

Comparison of Adequacy of Dialysis between Single-use and Reused Hemodialyzers in Patients on Maintenance Hemodialysis

Hamid

Dhrolia

Imtiaz

et al. 2019

J Coll Physicians Surg Pak

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“…Phenotypic methods developed recently seems to be both rapidness and cost. These methods are colorimetric methods (resazurin microplate method or resazurin tube test11, malachite green decolorization assay1213, nitrate reductase test)10, gradient diffusion method (E-test)14, microscopic observation drug susceptibility assay15, and thin-layer agar method16. In comparing these new methods with each other, they have advantage and disadvantage in performing and evaluation stages.…”

mentioning

confidence: 99%

Multicenter evaluation of crystal violet decolorization assay (CVDA) for rapid detection of isoniazid and rifampicin resistance in Mycobacterium tuberculosis

Çoban

Akbal

Biçmen

et al. 2016

Sci Rep

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The aim of this multicenter study was to evaluate the performance of the crystal violet decolorization assay (CVDA) for detection of multidrug resistant tuberculosis (MDR-TB). This study was performed in 11 centers in two phases. A total of 156 isolates were tested for INH and RIF resistance. In the phase I, 106 clinical isolates were tested in the Center 1–7. In the phase 2, 156 clinical isolates were tested in the center 1–6, center 8–11. Eighty six of 156 tested isolates were the same in phase I. Agreements were 96.2–96.8% for INH and 98.1–98.7% for RIF in the phase I-II, respectively. Mean time to obtain the results in the phase I was 14.3 ± 5.4 days. In the phase II, mean time to obtain the results was 11.6 ± 3.5 days. Test results were obtained within 14days for 62.3% (66/106) of isolates in the phase I and 81.4% (127/156) of isolates in the phase II. In conclusion, CVDA is rapid, reliable, inexpensive, and easy to perform for rapid detection of MDR-TB isolates. In addition, it could be adapted for drug susceptibility testing with all drugs both in developed and developing countries.

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The thin-layer agar method for direct phenotypic detection of multi- and extensively drug-resistant tuberculosis

Abstract: In low-resource settings, TLA can be applied for the rapid detection of resistance to INH, RMP and fluoroquinolones. Further studies are necessary to improve sensitivity to KM and further assess its performance for OFX and other drugs and its applicability in field conditions.

Cited by 13 publications

References 10 publications

Successful introduction of the Colour Test into inexperienced settings

Successful introduction of the Colour Test into inexperienced settings

Comparison of Adequacy of Dialysis between Single-use and Reused Hemodialyzers in Patients on Maintenance Hemodialysis

Multicenter evaluation of crystal violet decolorization assay (CVDA) for rapid detection of isoniazid and rifampicin resistance in Mycobacterium tuberculosis

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