Detection of embB codon 306 mutations in ethambutol resistant Mycobacterium tuberculosis directly from sputum samples: A low-cost, rapid approach

H. Rinder, K. T. Mieskes, E. Tortoli, E. Richter, M. Casal, M. Vaquero, E. Cambau, K. Feldmann, T. Löscher

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Substitutions of codon 306 in the gene embB are the most common mutations found in ethambutol resistant Mycobacterium tuberculosis. The characterization of these mutations has been hampered by the need for prior cultivation of the mycobacteria, or the need for DNA sequencing, or both. Here, we describe a simple and culture-independent technique to detect embB codon 306 mutations directly from sputum samples, requiring little more than a PCR machine and a simple agarose minigel. There is no need for labelled probes or DNA sequencing. In a preliminary test of feasibility, interpretable results were obtained from 21 of 24 selected sputum samples, 12 of which were determined to contain ethambutol resistant M. tuberculosis after culture. All of six samples with embB codon 306 mutations were correctly identified. Although an exact validation of this technique is beyond the scope of this technical report, we conclude from well-known embB codon 306 mutation prevalence figures that approximately one half of EMB resistant cases could already be predicted within 2 working days, with little equipment or hands-on time needed, instead of weeks required for conventional resistance testing.

Original languageEnglish
Pages (from-to)37-42
Number of pages6
JournalMolecular and Cellular Probes
Volume15
Issue number1
DOIs
Publication statusPublished - 2001

Fingerprint

Ethambutol
Sputum
Mycobacterium tuberculosis
Codon
Costs and Cost Analysis
Mutation
DNA Sequence Analysis
Culture Techniques
Mycobacterium
Sepharose
Equipment and Supplies
Polymerase Chain Reaction
Genes

Keywords

  • embB
  • Ethambutol
  • Mycobacterium tuberculosis
  • Resistance

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

Detection of embB codon 306 mutations in ethambutol resistant Mycobacterium tuberculosis directly from sputum samples : A low-cost, rapid approach. / Rinder, H.; Mieskes, K. T.; Tortoli, E.; Richter, E.; Casal, M.; Vaquero, M.; Cambau, E.; Feldmann, K.; Löscher, T.

In: Molecular and Cellular Probes, Vol. 15, No. 1, 2001, p. 37-42.

Research output: Contribution to journalArticle

Rinder, H, Mieskes, KT, Tortoli, E, Richter, E, Casal, M, Vaquero, M, Cambau, E, Feldmann, K & Löscher, T 2001, 'Detection of embB codon 306 mutations in ethambutol resistant Mycobacterium tuberculosis directly from sputum samples: A low-cost, rapid approach', Molecular and Cellular Probes, vol. 15, no. 1, pp. 37-42. https://doi.org/10.1006/mcpr.2000.0339
Rinder, H. ; Mieskes, K. T. ; Tortoli, E. ; Richter, E. ; Casal, M. ; Vaquero, M. ; Cambau, E. ; Feldmann, K. ; Löscher, T. / Detection of embB codon 306 mutations in ethambutol resistant Mycobacterium tuberculosis directly from sputum samples : A low-cost, rapid approach. In: Molecular and Cellular Probes. 2001 ; Vol. 15, No. 1. pp. 37-42.
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