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Ethambutol resistance testing by mutation detection.

R Johnson1, A M Jordaan, L Pretorius

  • 1Medical Research Council Centre for Molecular and Cellular Biology Medical Biochemistry and Faculty of Health Sciences, Stellenbosch University, Tygerberg, South Africa.

The International Journal of Tuberculosis and Lung Disease : the Official Journal of the International Union Against Tuberculosis and Lung Disease
|February 10, 2006
PubMed
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Chromosomal mutations in the embB gene are key indicators of ethambutol resistance in Mycobacterium tuberculosis. Many cases of EMB resistance were missed by standard laboratory tests, highlighting the need for improved diagnostic methods.

Area of Science:

  • Microbiology
  • Genetics
  • Infectious Diseases

Background:

  • Tuberculosis (TB) remains a significant global health challenge.
  • Drug resistance in Mycobacterium tuberculosis complicates treatment regimens.
  • Ethambutol (EMB) is a crucial first-line drug for TB treatment.

Purpose of the Study:

  • To identify specific chromosomal mutations conferring ethambutol resistance in Mycobacterium tuberculosis.
  • To evaluate the diagnostic accuracy of genotypic methods for EMB resistance detection.

Main Methods:

  • Analysis of embB gene mutations in 235 drug-resistant and 117 drug-susceptible M. tuberculosis isolates.
  • Comparison of genotypic findings with phenotypic EMB susceptibility testing (agar diffusion and BACTEC).
  • Utilized a novel one-step amplification refractory mutation system polymerase chain reaction (ARMS-PCR) for genotype detection.

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Main Results:

  • Mutations at embB codon 306 were identified in 20% of resistant isolates, significantly higher than in susceptible ones.
  • Phenotypic testing revealed that 91.4% of EMB resistance cases were missed by routine laboratory procedures.
  • EMB resistance was strongly associated with multidrug resistance (MDR), co-occurring with isoniazid and rifampicin resistance in 87.2% of cases.
  • The ARMS-PCR method demonstrated effectiveness in detecting EMB-resistant genotypes.

Conclusions:

  • Current diagnostic methods for EMB resistance in M. tuberculosis may miss a substantial number of resistant cases.
  • Improved diagnostic strategies, including genotypic analysis, are essential for accurate patient management.
  • Accurate EMB resistance testing is critical for guiding standardized treatment of multidrug-resistant TB (MDR-TB).