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Utility of the REBA MTB-rifa® assay for rapid detection of rifampicin resistant Mycobacterium Tuberculosis

BACKGROUND: Drug-resistant tuberculosis (TB), including resistance to both rifampicin (RIF) and isoniazid (INH) referred to as multidrug-resistant tuberculosis (MDR-TB), has become an increasing global threat in recent years. Effective management of patients infected with MDR-TB strains requires ide...

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Autores principales: Cho, Eunjin, Shamputa, Isdore Chola, Kwak, Hyun-Kyung, Lee, Jiim, Lee, Myungsun, Hwang, Soohee, Jeon, Doosoo, Kim, Cheon Tae, Cho, Sangnae, Via, Laura E, Barry 3rd, Clifton E, Lee, Jong Seok
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3852947/
https://www.ncbi.nlm.nih.gov/pubmed/24128118
http://dx.doi.org/10.1186/1471-2334-13-478
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author Cho, Eunjin
Shamputa, Isdore Chola
Kwak, Hyun-Kyung
Lee, Jiim
Lee, Myungsun
Hwang, Soohee
Jeon, Doosoo
Kim, Cheon Tae
Cho, Sangnae
Via, Laura E
Barry 3rd, Clifton E
Lee, Jong Seok
author_facet Cho, Eunjin
Shamputa, Isdore Chola
Kwak, Hyun-Kyung
Lee, Jiim
Lee, Myungsun
Hwang, Soohee
Jeon, Doosoo
Kim, Cheon Tae
Cho, Sangnae
Via, Laura E
Barry 3rd, Clifton E
Lee, Jong Seok
author_sort Cho, Eunjin
collection PubMed
description BACKGROUND: Drug-resistant tuberculosis (TB), including resistance to both rifampicin (RIF) and isoniazid (INH) referred to as multidrug-resistant tuberculosis (MDR-TB), has become an increasing global threat in recent years. Effective management of patients infected with MDR-TB strains requires identifying such patients by performing conventional drug-susceptibility testing (DST) on bacteria isolated from sputum, a process that can take up to 2 months. This delay in diagnosis can result in worsening and continued transmission of MDR-TB. Molecular methods that rely upon nucleic acid amplification of specific alleles known to be associated with resistance to specific drugs have been helpful in shortening the time to detect drug resistant TB. METHODS: We investigated the utility of the REBA MTB-Rifa®, a commercially available line probe assay (LPA) for detecting rifampicin (RIF) resistance in the RIF resistance-determining region (RRDR) of the rpoB gene. Altogether, 492 Mycobacterium tuberculosis (M. tuberculosis) clinical isolates and additional 228 smear- and culture-positive sputum samples with confirmed M. tuberculosis were collected from subjects with suspected MDR-TB in South Korea. The results were compared with conventional phenotypic DST and sequencing of the rpoB gene. RESULTS: A total of 215 of the 492 isolates were resistant to RIF by conventional DST, and of which 92.1% (198/215) were MDR-TB strains. The REBA MTB-Rifa® assay identified RIF resistance in 98.1% (211/215) of these isolates but failed to identify resistance in four phenotypically RIF resistant isolates. These four isolates lacked mutations in the RRDR but three were confirmed to be MDR-TB strains by sequencing. The sensitivity and specificity of this test for clinical isolates was thus 98.1% (211/215) and 100% (277/277), respectively. When applied directly to 228 smear positive sputum samples, the sensitivity and the specificity of REBA MTB-Rifa® assay was 100% (96/96, 132/132), respectively. CONCLUSIONS: These findings support the use of the REBA MTB-Rifa® assay for rapid detection of RIF resistance on clinical isolates and smear positive sputum samples. The results also suggest that RIF resistance is a good surrogate marker of MDR-TB in South Korea and the need to add more probes to other LPAs which can cover newly identified mutations relevant to RIF resistance.
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spelling pubmed-38529472013-12-07 Utility of the REBA MTB-rifa® assay for rapid detection of rifampicin resistant Mycobacterium Tuberculosis Cho, Eunjin Shamputa, Isdore Chola Kwak, Hyun-Kyung Lee, Jiim Lee, Myungsun Hwang, Soohee Jeon, Doosoo Kim, Cheon Tae Cho, Sangnae Via, Laura E Barry 3rd, Clifton E Lee, Jong Seok BMC Infect Dis Research Article BACKGROUND: Drug-resistant tuberculosis (TB), including resistance to both rifampicin (RIF) and isoniazid (INH) referred to as multidrug-resistant tuberculosis (MDR-TB), has become an increasing global threat in recent years. Effective management of patients infected with MDR-TB strains requires identifying such patients by performing conventional drug-susceptibility testing (DST) on bacteria isolated from sputum, a process that can take up to 2 months. This delay in diagnosis can result in worsening and continued transmission of MDR-TB. Molecular methods that rely upon nucleic acid amplification of specific alleles known to be associated with resistance to specific drugs have been helpful in shortening the time to detect drug resistant TB. METHODS: We investigated the utility of the REBA MTB-Rifa®, a commercially available line probe assay (LPA) for detecting rifampicin (RIF) resistance in the RIF resistance-determining region (RRDR) of the rpoB gene. Altogether, 492 Mycobacterium tuberculosis (M. tuberculosis) clinical isolates and additional 228 smear- and culture-positive sputum samples with confirmed M. tuberculosis were collected from subjects with suspected MDR-TB in South Korea. The results were compared with conventional phenotypic DST and sequencing of the rpoB gene. RESULTS: A total of 215 of the 492 isolates were resistant to RIF by conventional DST, and of which 92.1% (198/215) were MDR-TB strains. The REBA MTB-Rifa® assay identified RIF resistance in 98.1% (211/215) of these isolates but failed to identify resistance in four phenotypically RIF resistant isolates. These four isolates lacked mutations in the RRDR but three were confirmed to be MDR-TB strains by sequencing. The sensitivity and specificity of this test for clinical isolates was thus 98.1% (211/215) and 100% (277/277), respectively. When applied directly to 228 smear positive sputum samples, the sensitivity and the specificity of REBA MTB-Rifa® assay was 100% (96/96, 132/132), respectively. CONCLUSIONS: These findings support the use of the REBA MTB-Rifa® assay for rapid detection of RIF resistance on clinical isolates and smear positive sputum samples. The results also suggest that RIF resistance is a good surrogate marker of MDR-TB in South Korea and the need to add more probes to other LPAs which can cover newly identified mutations relevant to RIF resistance. BioMed Central 2013-10-15 /pmc/articles/PMC3852947/ /pubmed/24128118 http://dx.doi.org/10.1186/1471-2334-13-478 Text en Copyright © 2013 Cho et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Cho, Eunjin
Shamputa, Isdore Chola
Kwak, Hyun-Kyung
Lee, Jiim
Lee, Myungsun
Hwang, Soohee
Jeon, Doosoo
Kim, Cheon Tae
Cho, Sangnae
Via, Laura E
Barry 3rd, Clifton E
Lee, Jong Seok
Utility of the REBA MTB-rifa® assay for rapid detection of rifampicin resistant Mycobacterium Tuberculosis
title Utility of the REBA MTB-rifa® assay for rapid detection of rifampicin resistant Mycobacterium Tuberculosis
title_full Utility of the REBA MTB-rifa® assay for rapid detection of rifampicin resistant Mycobacterium Tuberculosis
title_fullStr Utility of the REBA MTB-rifa® assay for rapid detection of rifampicin resistant Mycobacterium Tuberculosis
title_full_unstemmed Utility of the REBA MTB-rifa® assay for rapid detection of rifampicin resistant Mycobacterium Tuberculosis
title_short Utility of the REBA MTB-rifa® assay for rapid detection of rifampicin resistant Mycobacterium Tuberculosis
title_sort utility of the reba mtb-rifa® assay for rapid detection of rifampicin resistant mycobacterium tuberculosis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3852947/
https://www.ncbi.nlm.nih.gov/pubmed/24128118
http://dx.doi.org/10.1186/1471-2334-13-478
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