Pyrosequencing for rapid detection of Mycobacterium tuberculosis second-line drugs and ethambutol resistance

Diagn Microbiol Infect Dis. 2015 Nov;83(3):263-9. doi: 10.1016/j.diagmicrobio.2015.07.004. Epub 2015 Jul 11.

Abstract

The aim of this work was to study the diagnostic accuracy of pyrosequencing to detect resistance to fluoroquinolones, kanamycin, amikacin, capreomycin, and ethambutol (EMB) in Mycobacterium tuberculosis clinical strains. One hundred four clinical isolates previously characterized by BACTEC 460TB/MGIT 960 were included. Specific mutations were targeted in gyrA, rrs, eis promoter, and embB. When there was a discordant result between BACTEC and pyrosequencing, Genotype MTBDRsl (Hain Lifescience, Nehren, Germany) was performed. Sensitivity and specificity of pyrosequencing were 70.6% and 100%, respectively, for fluoroquinolones; 93.3% and 81.7%, respectively, for kanamycin; 94.1% and 95.9%, respectively, for amikacin; 90.0% and 100%, respectively, for capreomycin; and 64.8% and 87.8%, respectively, for EMB. This study shows that pyrosequencing may be a useful tool for making early decisions regarding second-line drugs and EMB resistance. However, for a correct management of patients with suspected extensively drug-resistant tuberculosis, susceptibility results obtained by molecular methods should be confirmed by a phenotypic method.

Keywords: Ethambutol; Extensively drug-resistant tuberculosis; Fluoroquinolones; Injectable drugs; Molecular diagnostic testing; Tuberculosis.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antitubercular Agents / pharmacology*
  • Drug Resistance, Bacterial*
  • Genotyping Techniques / methods*
  • Humans
  • Microbial Sensitivity Tests / methods*
  • Mycobacterium tuberculosis / drug effects
  • Mycobacterium tuberculosis / genetics*
  • Sensitivity and Specificity
  • Sequence Analysis, DNA / methods*
  • Time Factors

Substances

  • Antitubercular Agents