Cloning and nucleotide sequence of Mycobacterium tuberculosis gyrA and gyrB genes and detection of quinolone resistance mutations

H. E. Takiff, L. Salazar, C. Guerrero, W. Philipp, Mun Huang Wai Mun Huang, B. Kreiswirth, S. T. Cole, W. R. Jacobs, A. Telenti

Research output: Contribution to journalArticle

Abstract

The emergence of multidrug-resistant strains of Mycobacterium tuberculosis has resulted in increased interest in the fluoroquinolones (FQs) as antituberculosis agents. To investigate the frequency and mechanisms of FQ resistance in M. tuberculosis, we cloned and sequenced the wild-type gyrA and gyrB genes, which encode the A and B subunits of the DNA gyrase, respectively; DNA gyrase is the main target of the FQs. On the basis of the sequence information, we performed DNA amplification for sequencing and single-strand conformation polymorphism analysis to examine the presumed quinolone resistance regions of gyrA and gyrB from reference strains (n = 4) and clinical isolates (n = 55). Mutations in codons of gyrA analogous to those described in other FQ-resistant bacteria were identified in all isolates (n = 14) for which the ciprofloxacin MIC was >2 μg/ml. In addition, we selected ciprofloxacin-resistant mutants of Mycobacterium bovis BCG and M. tuberculosis Erdman and H37ra. Spontaneously resistant mutants developed at a frequency of 1 in 107 to 108 at ciprofloxacin concentrations of 2 μg/ml, but no primary resistant colonies were selected at higher ciprofloxacin concentrations. Replating of those first-step mutants selected for mutants with high levels of resistance which harbored gyrA mutations similar to those found among clinical FQ-resistant isolates. The gyrA and gyrB sequence information will facilitate analysis of the mechanisms of resistance to drugs which target the gyrase and the implementation of rapid strategies for the estimation of FQ susceptibility in clinical M. tuberculosis isolates.

Original languageEnglish (US)
Pages (from-to)773-780
Number of pages8
JournalAntimicrobial Agents and Chemotherapy
Volume38
Issue number4
DOIs
StatePublished - Jan 1 1994
Externally publishedYes

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Molecular Sequence Data
Bacterial Genes
Type II DNA Topoisomerase
Quinolones
Fluoroquinolones
Molecular Cloning
Ciprofloxacin
Microbial Drug Resistance
Mycobacterium bovis
Mycobacterium tuberculosis
Organism Cloning
Tuberculosis
DNA Gyrase
Polymerase Chain Reaction
Mutation
Genes
DNA Sequence Analysis
Drug Resistance
Codon
Bacteria

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)
  • Infectious Diseases

Cite this

Takiff, H. E., Salazar, L., Guerrero, C., Philipp, W., Wai Mun Huang, M. H., Kreiswirth, B., ... Telenti, A. (1994). Cloning and nucleotide sequence of Mycobacterium tuberculosis gyrA and gyrB genes and detection of quinolone resistance mutations. Antimicrobial Agents and Chemotherapy, 38(4), 773-780. https://doi.org/10.1128/AAC.38.4.773

Cloning and nucleotide sequence of Mycobacterium tuberculosis gyrA and gyrB genes and detection of quinolone resistance mutations. / Takiff, H. E.; Salazar, L.; Guerrero, C.; Philipp, W.; Wai Mun Huang, Mun Huang; Kreiswirth, B.; Cole, S. T.; Jacobs, W. R.; Telenti, A.

In: Antimicrobial Agents and Chemotherapy, Vol. 38, No. 4, 01.01.1994, p. 773-780.

Research output: Contribution to journalArticle

Takiff, HE, Salazar, L, Guerrero, C, Philipp, W, Wai Mun Huang, MH, Kreiswirth, B, Cole, ST, Jacobs, WR & Telenti, A 1994, 'Cloning and nucleotide sequence of Mycobacterium tuberculosis gyrA and gyrB genes and detection of quinolone resistance mutations', Antimicrobial Agents and Chemotherapy, vol. 38, no. 4, pp. 773-780. https://doi.org/10.1128/AAC.38.4.773
Takiff, H. E. ; Salazar, L. ; Guerrero, C. ; Philipp, W. ; Wai Mun Huang, Mun Huang ; Kreiswirth, B. ; Cole, S. T. ; Jacobs, W. R. ; Telenti, A. / Cloning and nucleotide sequence of Mycobacterium tuberculosis gyrA and gyrB genes and detection of quinolone resistance mutations. In: Antimicrobial Agents and Chemotherapy. 1994 ; Vol. 38, No. 4. pp. 773-780.
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