Optimization of the microscopic observation drug susceptibility assay for four first-line drugs using Mycobacterium tuberculosis reference strains and clinical isolates

J Microbiol Methods. 2014 Jun:101:44-8. doi: 10.1016/j.mimet.2014.03.014. Epub 2014 Apr 6.

Abstract

Objective: The aim of this study is to determine the appropriate cut-off value and turnaround time of the microscopic observation drug susceptibility assay (MODS) for isoniazid (INH), rifampicin (RMP), streptomycin (STR), and ethambutol (EMB).

Design: A total of 39 Mycobacterium tuberculosis strains with confirmed drug susceptibility (reference strains) were tested with a range of drug concentrations to determine the optimal cut-off values for INH, RMP, STR, and EMB by MODS. Standard drug susceptibility testing (DST) results were evaluated relative to the Löwenstein-Jensen (L-J) proportion method. Following which, the performance of MODS was evaluated again using 36 sputum samples from patients with tuberculosis (TB) using the cut-off values determined in the aforementioned process.

Results: With 39 reference strains, DST identified the following cut-off values: 0.8μg/ml INH (sensitivity, 96.0%; specificity, 92.9%), 2.0μg/ml RMP (sensitivity, 100%; specificity, 95.5%), 4.0μg/ml STR (sensitivity, 90.5%; specificity, 93.8%), and 4.0μg/ml EMB (sensitivity, 100%; specificity, 91.7%). When these cut-off values were used to analyze the 36 clinical isolates, the sensitivity and specificity of MODS were 100% and 93.1% for INH, 100% and 93.8% for RMP, 87.5% and 96.4% for STR, and 100% and 88.2% for EMB, respectively. The turnaround time for these clinical specimens was 9.0days by MODS (95% CI: 5.3-12.7), compared with 11.7days (95% CI: 9.5-13.9) for smear negative specimens.

Conclusion: Our study identified the optimal cut-off values of the four first-line drugs for MODS based on a wide concentration range. With the optimal cut-off values determined in this study, MODS showed high discriminatory efficiency for DST. This study also demonstrated that MODS is useful for rapid diagnosis of drug-resistant TB even for a smear negative specimen, despite the fact that it generally uses smear positive specimens as direct DST.

Keywords: Drug susceptibility testing; MODS; Smear negative; Turnaround time.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antitubercular Agents / pharmacology*
  • Ethambutol / pharmacology
  • Humans
  • Isoniazid / pharmacology
  • Microbial Sensitivity Tests / methods*
  • Mycobacterium tuberculosis / drug effects*
  • Rifampin / pharmacology
  • Sensitivity and Specificity
  • Sputum / microbiology
  • Streptomycin / pharmacology
  • Time Factors
  • Tuberculosis, Pulmonary / microbiology*

Substances

  • Antitubercular Agents
  • Ethambutol
  • Isoniazid
  • Rifampin
  • Streptomycin