Evaluation of the Quantamatrix Multiplexed Assay Platform system for simultaneous detection of Mycobacterium tuberculosis and the rifampicin resistance gene using cultured mycobacteria

Hye young Wang, Young Uh, Seoyong Kim, Tae sun Shim, Hyeyoung Lee

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4 Citations (Scopus)

Abstract

Background The differentiation of Mycobacterium tuberculosis complex (MTBC) from non-tuberculous mycobacteria (NTM) is of primary importance for infection control and the selection of anti-tuberculosis drugs. Up to date data on rifampicin (RIF)-resistant tuberculosis (TB) is essential for the early management of multidrug-resistant TB. The aim of this study was to evaluate the usefulness of a newly developed multiplexed, bead-based bioassay (Quantamatrix Multiplexed Assay Platform, QMAP) for the rapid differentiation of 23 Mycobacterium species including MTBC and RIF-resistant strains. Methods A total of 314 clinical Mycobacterium isolates cultured from respiratory specimens were used in this study. Results The sensitivity and specificity of the QMAP system for Mycobacterium species were 100% (95% CI 99.15–100%, p < 0.0001) and 97.8% (95% CI 91.86–99.87%, p < 0.0001), respectively. The results of conventional drug susceptibility testing and the QMAP Dual-ID assay were completely concordant for all clinical isolates (100%, 95% CI 98.56–100%). Out of 223 M. tuberculosis (MTB) isolates, 196 were pan-susceptible and 27 were resistant to RIF according to QMAP results. All of the mutations in the RIF resistance-determining region detected by the QMAP system were confirmed by rpoB sequence analysis and a REBA MTB-Rifa reverse blot hybridization assay. The majority of the mutations (n = 26, 96.3%), including those missing wild-type probe signals, were located in three codons (529–534, 524–529, and 514–520), and 17 (65.4%) of these mutations were detected by three mutation probes (531TTG, 526TAC, and 516GTC). Conclusions The entire QMAP system assay takes about 3 h to complete, while results from the culture-based conventional method can take up to 48–72 h. Although improvements to the QMAP system are needed for direct respiratory specimens, it may be useful for rapid screening, not only to identify and accurately discriminate MTBC from NTM, but also to identify RIF-resistant MTB strains in positive culture samples.

Original languageEnglish
Pages (from-to)107-113
Number of pages7
JournalInternational Journal of Infectious Diseases
Volume61
DOIs
Publication statusPublished - 2017 Aug 1

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Rifampin
Mycobacterium
Mycobacterium tuberculosis
Genes
Mutation
Tuberculosis
Multidrug-Resistant Tuberculosis
Infection Control
Codon
Biological Assay
Pharmaceutical Preparations
Sequence Analysis
Sensitivity and Specificity

All Science Journal Classification (ASJC) codes

  • Microbiology (medical)
  • Infectious Diseases

Cite this

@article{c6e88c3294354cde8f589490c1a033a2,
title = "Evaluation of the Quantamatrix Multiplexed Assay Platform system for simultaneous detection of Mycobacterium tuberculosis and the rifampicin resistance gene using cultured mycobacteria",
abstract = "Background The differentiation of Mycobacterium tuberculosis complex (MTBC) from non-tuberculous mycobacteria (NTM) is of primary importance for infection control and the selection of anti-tuberculosis drugs. Up to date data on rifampicin (RIF)-resistant tuberculosis (TB) is essential for the early management of multidrug-resistant TB. The aim of this study was to evaluate the usefulness of a newly developed multiplexed, bead-based bioassay (Quantamatrix Multiplexed Assay Platform, QMAP) for the rapid differentiation of 23 Mycobacterium species including MTBC and RIF-resistant strains. Methods A total of 314 clinical Mycobacterium isolates cultured from respiratory specimens were used in this study. Results The sensitivity and specificity of the QMAP system for Mycobacterium species were 100{\%} (95{\%} CI 99.15–100{\%}, p < 0.0001) and 97.8{\%} (95{\%} CI 91.86–99.87{\%}, p < 0.0001), respectively. The results of conventional drug susceptibility testing and the QMAP Dual-ID assay were completely concordant for all clinical isolates (100{\%}, 95{\%} CI 98.56–100{\%}). Out of 223 M. tuberculosis (MTB) isolates, 196 were pan-susceptible and 27 were resistant to RIF according to QMAP results. All of the mutations in the RIF resistance-determining region detected by the QMAP system were confirmed by rpoB sequence analysis and a REBA MTB-Rifa reverse blot hybridization assay. The majority of the mutations (n = 26, 96.3{\%}), including those missing wild-type probe signals, were located in three codons (529–534, 524–529, and 514–520), and 17 (65.4{\%}) of these mutations were detected by three mutation probes (531TTG, 526TAC, and 516GTC). Conclusions The entire QMAP system assay takes about 3 h to complete, while results from the culture-based conventional method can take up to 48–72 h. Although improvements to the QMAP system are needed for direct respiratory specimens, it may be useful for rapid screening, not only to identify and accurately discriminate MTBC from NTM, but also to identify RIF-resistant MTB strains in positive culture samples.",
author = "Wang, {Hye young} and Young Uh and Seoyong Kim and Shim, {Tae sun} and Hyeyoung Lee",
year = "2017",
month = "8",
day = "1",
doi = "10.1016/j.ijid.2017.06.008",
language = "English",
volume = "61",
pages = "107--113",
journal = "International Journal of Infectious Diseases",
issn = "1201-9712",
publisher = "Elsevier",

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TY - JOUR

T1 - Evaluation of the Quantamatrix Multiplexed Assay Platform system for simultaneous detection of Mycobacterium tuberculosis and the rifampicin resistance gene using cultured mycobacteria

AU - Wang, Hye young

AU - Uh, Young

AU - Kim, Seoyong

AU - Shim, Tae sun

AU - Lee, Hyeyoung

PY - 2017/8/1

Y1 - 2017/8/1

N2 - Background The differentiation of Mycobacterium tuberculosis complex (MTBC) from non-tuberculous mycobacteria (NTM) is of primary importance for infection control and the selection of anti-tuberculosis drugs. Up to date data on rifampicin (RIF)-resistant tuberculosis (TB) is essential for the early management of multidrug-resistant TB. The aim of this study was to evaluate the usefulness of a newly developed multiplexed, bead-based bioassay (Quantamatrix Multiplexed Assay Platform, QMAP) for the rapid differentiation of 23 Mycobacterium species including MTBC and RIF-resistant strains. Methods A total of 314 clinical Mycobacterium isolates cultured from respiratory specimens were used in this study. Results The sensitivity and specificity of the QMAP system for Mycobacterium species were 100% (95% CI 99.15–100%, p < 0.0001) and 97.8% (95% CI 91.86–99.87%, p < 0.0001), respectively. The results of conventional drug susceptibility testing and the QMAP Dual-ID assay were completely concordant for all clinical isolates (100%, 95% CI 98.56–100%). Out of 223 M. tuberculosis (MTB) isolates, 196 were pan-susceptible and 27 were resistant to RIF according to QMAP results. All of the mutations in the RIF resistance-determining region detected by the QMAP system were confirmed by rpoB sequence analysis and a REBA MTB-Rifa reverse blot hybridization assay. The majority of the mutations (n = 26, 96.3%), including those missing wild-type probe signals, were located in three codons (529–534, 524–529, and 514–520), and 17 (65.4%) of these mutations were detected by three mutation probes (531TTG, 526TAC, and 516GTC). Conclusions The entire QMAP system assay takes about 3 h to complete, while results from the culture-based conventional method can take up to 48–72 h. Although improvements to the QMAP system are needed for direct respiratory specimens, it may be useful for rapid screening, not only to identify and accurately discriminate MTBC from NTM, but also to identify RIF-resistant MTB strains in positive culture samples.

AB - Background The differentiation of Mycobacterium tuberculosis complex (MTBC) from non-tuberculous mycobacteria (NTM) is of primary importance for infection control and the selection of anti-tuberculosis drugs. Up to date data on rifampicin (RIF)-resistant tuberculosis (TB) is essential for the early management of multidrug-resistant TB. The aim of this study was to evaluate the usefulness of a newly developed multiplexed, bead-based bioassay (Quantamatrix Multiplexed Assay Platform, QMAP) for the rapid differentiation of 23 Mycobacterium species including MTBC and RIF-resistant strains. Methods A total of 314 clinical Mycobacterium isolates cultured from respiratory specimens were used in this study. Results The sensitivity and specificity of the QMAP system for Mycobacterium species were 100% (95% CI 99.15–100%, p < 0.0001) and 97.8% (95% CI 91.86–99.87%, p < 0.0001), respectively. The results of conventional drug susceptibility testing and the QMAP Dual-ID assay were completely concordant for all clinical isolates (100%, 95% CI 98.56–100%). Out of 223 M. tuberculosis (MTB) isolates, 196 were pan-susceptible and 27 were resistant to RIF according to QMAP results. All of the mutations in the RIF resistance-determining region detected by the QMAP system were confirmed by rpoB sequence analysis and a REBA MTB-Rifa reverse blot hybridization assay. The majority of the mutations (n = 26, 96.3%), including those missing wild-type probe signals, were located in three codons (529–534, 524–529, and 514–520), and 17 (65.4%) of these mutations were detected by three mutation probes (531TTG, 526TAC, and 516GTC). Conclusions The entire QMAP system assay takes about 3 h to complete, while results from the culture-based conventional method can take up to 48–72 h. Although improvements to the QMAP system are needed for direct respiratory specimens, it may be useful for rapid screening, not only to identify and accurately discriminate MTBC from NTM, but also to identify RIF-resistant MTB strains in positive culture samples.

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