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Mass-spectrometry analysis of genetic markers of S. pneumoniae resistance to β-lactam antibiotics

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Abstract

Despite the growing level of resistance to Streptococcus pneumoniae infections, β-lactam antibiotics remain the drugs of choice treating these infections. The resistance of S. pneumoniae to these preparations is mediated by modifications of penicillin-binding proteins (PBPs), which are the targets of antibiotics action. The new approach to detecting mutations in the PBP 1A, 2B, and 2X genes based on the minise-quencing reaction followed by matrix-assisted laser desorption/ionization time of flight (MALDI-ToF) mass spectrometry has been developed in the present study. The evaluation of the prevalence of these mutations in clinical S. pneumoniae isolates (n = 194) with different levels of susceptibility to beta-lactam antibiotics has been performed. In summary, 24 different combinations of mutations (genotypes) have been detected in PBPs. All penicillin-susceptible isolates (n = 49, MIC ≤ 0.06 μg/ml) were characterized by the absence of mutations in all analyzed loci. In PBPs, mutations were detected in 133 (91.7%) out of 145 S. pneumoniae isolates with reduced susceptibility to penicillin (MIC > 0.06 μg/ml), which indicates the high diagnostic sensitivity of this approach. Isolates with MIC → 4 μg/ml (n = 20) possessed multiple mutations in all analyzed genes, which confirms the cumulative effects of the formation of penicillin resistance. At the same time, no association between the presence of mutations in PBP genes and decreased susceptibility to cefotaxime was shown, which makes it possible to suggest significant differences in molecular mechanisms of penicillins and cephalosporins resistance. The suggested method of S. pneumoniae genotyping is appropriate for the individual screening of the susceptibility of isolates to penicillin and the molecular monitoring of the resistance determinants in population.

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Authors and Affiliations

  1. National Research Center for Medical Products Quality Control, Moscow, Russia

    T. A. Savinova

  2. Research Institute of Physicochemical Medicine, Moscow, Russia

    E. N. Il’ina

  3. Research Institute of Childhood Infections, St. Petersburg, Russia

    S. V. Sidorenko

Authors
  1. T. A. Savinova
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  2. E. N. Il’ina
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  3. S. V. Sidorenko
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Additional information

Original Russian Text © T.A. Savinova, E.N. Il’ina, S.V. Sidorenko, 2010, published in Molekulyarnaya Genetika, Mikrobiologiya i Virusologiya, 2010, No. 3, pp. 16–25.

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Savinova, T.A., Il’ina, E.N. & Sidorenko, S.V. Mass-spectrometry analysis of genetic markers of S. pneumoniae resistance to β-lactam antibiotics. Mol. Genet. Microbiol. Virol. 25, 106–117 (2010). https://doi.org/10.3103/S0891416810030043

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  • DOI: https://doi.org/10.3103/S0891416810030043

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