Single-nucleotide polymorphism-based differentiation and drug resistance detection in Mycobacterium tuberculosis from isolates or directly from sputum

Catherine Arnold*, L. Westland, G. Mowat, A. Underwood, J. Magee, Saheer Gharbia

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

63 Citations (Scopus)

Abstract

The rapid technique of pyrosequencing was used to examine 123 samples (in the form of DNA extracts and inactivated sputum) of Mycobacterium spp. Of 99 Mycobacterium tuberculosis samples investigated for single-nucleotide polymorphisms (SNPs), 68% of isoniazid-resistant isolates analysed had an AGC → ACC mutation in katG at codon 315, resulting in the Ser → Thr substitution associated previously with isoniazid resistance. Of the rifampicin-resistant isolates, 92% showed SNPs in rpoB at codons 516, 531 or 526. Inactivated sputum samples and DNA extracts could both be analysed by pyrosequencing, and the method was able to differentiate rapidly between the closely related species of the M. tuberculosis complex (M. tuberculosis, Mycobacterium bovis, Mycobacterium africanum, Mycobacterium canetti and Mycobacterium microti), except between M. tuberculosis, M. canetti and one of two M. africanum strains. This low-cost, high-throughput technique could be used as a rapid screen for drug resistance and as a replacement for some of the time-consuming tests used currently for species identification.

Original languageEnglish
Pages (from-to)122-130
Number of pages9
JournalClinical Microbiology and Infection
Volume11
Issue number2
DOIs
Publication statusPublished - Feb 2005

Keywords

  • Antibiotic susceptibility testing
  • Identification
  • Mycobacterium tuberculosis complex
  • Pyrosequencing
  • Resistance

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