A Multi-Site Validation in India of the Line Probe Assay for the Rapid Diagnosis of Multi-Drug Resistant Tuberculosis Directly from Sputum Specimens

Raizada, Neeraj; Sachdeva, Kuldeep Singh; Chauhan, D. S.; Malhotra, Bharti; Reddy, Kishore; Dave, Paresh; Mundade, Yamuna; Patel, P.G.; Ramachandran, Ranjani; Das, Ram; Solanki, Rajesh; Wares, D F; Sahu, Suvanand; O’Brien, Rick; Paramasivan, C. N.; Dewan, Puneet

doi:10.1371/journal.pone.0088626

Cited by 42 publications

(33 citation statements)

References 17 publications

(16 reference statements)

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“…Recent studies have found the MTBDR plus assay to be an effective and feasible diagnostic tool for MDR-TB screening in endemic regions [12, 13, 14, 24, 42]. While the sensitivity of the molecular diagnostic test for detection of INH resistance in our study was slightly lower than that for RIF and MDR detection (compared to conventional methods), it was analogous to that of previously reported studies [25, 31, 32].…”

Section: Discussionsupporting

confidence: 86%

“…LPA indeterminate have been previously reported to be related to smear- and culture-status of the evaluated samples, and generally occur in 1.4–19.2% of samples, depending on sample type (EPTB v. PTB), [40] with un-interpretable reads only occurring in 6% of smear positive sputum samples upon test repeat [42]. Our study found 12.4% of results to be NI without test repeat.…”

Section: Discussionsupporting

confidence: 50%

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Redefining MTBDR<I>plus</I> test results: what do indeterminate results actually mean?

Nikam¹,

Patel²,

Sadani³

et al. 2016

int j tuberc lung dis

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Early diagnosis of drug-resistant tuberculosis is critical in order to establish appropriate drug treatment regimens and to prevent the further transmission of resistant strains. Line probe assays (LPAs) are promising rapid diagnostics for use in low income settings, and can be used to screen smear-positive sputum samples for resistance to rifampicin (RIF) and isoniazid (INH) in as little as 1–2 days. Though the diagnostic performance of these assays has been well evaluated, little research has elucidated the true nature of indeterminate LPA results, or assessed the ability of these assays to perform on a wide range of clinical samples We evaluated the performance of the commercially available Genotype MTBDRplus LPA against conventional MGIT 960 culture and drug susceptibility testing (DST) results for 308 pulmonary (PTB) and 32 extrapulmonary (EPTB) tuberculosis samples. Invalid LPA results (defined as missing the MTB identification band) were obtained for 18 PTB samples, which were excluded from further analysis. The sensitivity and specificity of the Genotype MTBDRplus assay for multidrug-resistant tuberculosis (MDR-TB), based upon the results obtained for the remaining 322 samples, was 95.2% and 95.1%, respectively. 13.7% (40/290) of the PTB samples were not interpretable, or indeterminate, by LPA (defined as having the absence of both wild type and corresponding mutation bands) for INH and/or RIF and were further evaluated by pyrosequencing (PSQ). Contrary to standard LPA interpretation, INH and RIF susceptibility were confirmed by both DST and PSQ in 7.5% (3/40) and 27.5% (11/40) of indeterminate samples, respectively. When LPA test results were not interpretable in this study, PSQ was found to be a valuable and rapid technique to resolve discrepancies.

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Section: Discussionsupporting

confidence: 86%

Section: Discussionsupporting

confidence: 50%

Redefining MTBDR<I>plus</I> test results: what do indeterminate results actually mean?

Nikam¹,

Patel²,

Sadani³

et al. 2016

int j tuberc lung dis

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“…With the introduction of LPA for the rapid diagnosis of drug-resistant TB, there has been a significant reduction in time to initiation of treatment in MDR suspect cases7. A multisite validation study from India found LPA to be a sensitive and specific tool for the detection of rifampicin resistance in AFB smear-positive sputum specimens8, though, in a study from Punjab, India, 2.7 per cent specimens were detected as invalid9.…”

Section: Discussionmentioning

confidence: 99%

Line probe assay for detection of Mycobacterium tuberculosis complex: An experience from Central India

et al. 2017

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Background & objectives:Mycobacterium tuberculosis complex may sometimes not be detected in sputum samples of suspected multidrug-resistant tuberculosis (MDR-TB) patients by line probe assay (LPA) even though they are smear positive for acid-fast bacilli (AFB). This retrospective analysis was attempted to understand and document our experience with LPA for detection of M. tuberculosis complex and diagnosis of MDR-TB under programmatic conditions.Methods:One thousand two hundred and ninety four sputum samples of MDR-TB suspects that were smear positive for AFB, and received from February to November 2013, were tested by LPA for the presence of M. tuberculosis complex and resistance to isoniazid (INH) and rifampicin as per the diagnostic mandate of an accredited reference laboratory. As per the mandate, those samples that were negative for M. tuberculosis complex were cultured, and the growth again tested by LPA. A retrospective analysis of the results was carried out.Results:M. tuberculosis complex could be detected in 1217 (94.04%) but not in 77 (5.9%) of smear-positive sputum samples. Of the 1217 positive samples, 232 (19.1%) were MDR, 130 (10.6%) were rifampicin monoresistant and 101 (8.3%) were INH monoresistant. Seven hundred and fifty four (61.9%) strains were found to be pansensitive. Overall, 5.1 per cent of the sputum samples were negative for M. tuberculosis complex by LPA and culture. In at least 10 (0.77%) sputum samples smear positive for AFB, M. tuberculosis complex could not be identified by LPA though M. tuberculosis was present, as evidenced by culture positivity.Interpretation & conclusions:LPA is a robust technique for diagnosis of drug-resistant TB that has provided the basis for rapid and effective control of drug-resistant TB in India. While the reasons for concomitantly negative LPA and culture results of smear-positive sputum samples from MDR-TB suspects may be many, the possible presence of non-tubercular mycobacteria in these samples and the likelihood of inappropriate therapy in these patients cannot be ruled out. Addition of culture to the diagnostic algorithm may enhance the diagnostic yield.

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“…La prueba GenoType MTBDRplus ® es útil y puede proporcionar resultados en ocho horas, lo que permite un diagnóstico rápido de la tuberculosis multirresistente, y favorece el inicio temprano y específico del tratamiento contra la tuberculosis (12,13). En nuestro estudio hubo una concordancia perfecta (kappa=1) entre los resultados de la secuenciación y los de la prueba GenoTypeMTBDRplus ® para la detección de mutaciones relacionadas con la resistencia a isoniacida.…”

Section: Discussionunclassified

GenoType® MTBDRplus 1.0 para detección de resistencia cruzada entre isoniazida y etionamida en aislamientos de Mycobacterium tuberculosis multifármaco-resistente

Rueda¹,

Realpe²,

Mejía³

et al. 2015

biomedica

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Contribución de los autores:Johana Rueda: ejecución del proyecto, análisis e interpretación de los datos y redacción del manuscrito Teresa Realpe: análisis e interpretación de los datos Gloria Mejía y Elsa Zapata: aplicación de la metodología Jaime Robledo: diseño y gestión del proyecto Todos los autores participaron en la asesoría, la revisión y la aprobación del artículo. Introducción. Una parte de los aislamientos de Mycobacterium tuberculosis multirresistente también presenta resistencia a la etionamida. Es importante determinar si la resistencia a la isoniacida es independiente o se cruza con la resistencia a la etionamida, ya que si sucede lo segundo habría que reevaluar el tratamiento antituberculoso de segunda línea. La prueba molecular GenoType MTBDRplus ® detecta las mutaciones asociadas con la resistencia a isoniacida y podría detectar la resistencia cruzada a la etionamida. Objetivo. Evaluar la prueba GenoType MTBDRplus ® y comparar su desempeño con el de la secuenciación, en la detección de mutaciones en el gen katG y en el promotor inhA en aislamientos clínicos de M. tuberculosis multirresistente. Materiales y métodos. Se utilizaron el estuche comercial GenoType MTBDRplus 1.0 ® y la secuenciación para evaluar mutaciones en el gen katG y en el promotor inhA en 30 aislamientos de M. tuberculosis multirresistente con resistencia a la etionamida. La cepa de laboratorio H37Rv y tres aislamientos sensibles a los medicamentos de primera línea, sirvieron de control. Resultados. Al comparar los resultados de la secuenciación y de la prueba GenoType MTBDRplus ® , el índice kappa fue de 1. Todos los aislamientos resistentes a la isoniacida y la etionamida tenían las mutaciones detectadas con la prueba GenoTypeMTBDRplus ® en el gen katG, y 40 % de ellos, las detectadas en el promotor inhA. Mediante secuenciación se encontraron, además, mutaciones en katG en posiciones diferentes a las detectadas por la prueba GenoType MTBDRplus ® . GenoType Conclusión. La prueba GenoTypeMTBDRplus® tiene la capacidad de detectar rápidamente la resistencia a isoniacida. Además, los resultados del estudio sugieren que también podría utilizarse como prueba de tamización para detectar la resistencia cruzada a etionamida.Palabras clave: Mycobacterium tuberculosis, tuberculosis resistente a múltiples medicamentos, etionamida, isoniacida, análisis de secuencia, mutación.

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A Multi-Site Validation in India of the Line Probe Assay for the Rapid Diagnosis of Multi-Drug Resistant Tuberculosis Directly from Sputum Specimens

Cited by 42 publications

References 17 publications

Redefining MTBDR<I>plus</I> test results: what do indeterminate results actually mean?

Redefining MTBDR<I>plus</I> test results: what do indeterminate results actually mean?

Line probe assay for detection of Mycobacterium tuberculosis complex: An experience from Central India

GenoType® MTBDRplus 1.0 para detección de resistencia cruzada entre isoniazida y etionamida en aislamientos de Mycobacterium tuberculosis multifármaco-resistente

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