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Phenotypic detection of clinical isolates of Haemophilus influenzae with altered penicillin-binding protein 3

  • A. Aguirre-Quiñonero
  • I. C. Pérez del Molino
  • C. García de la Fuente
  • M. C. Sanjuán
  • J. Agüero
  • L. Martínez-Martínez
Original Article

Abstract

The aims of this study were to determine the correlation of mutations in the ftsI gene (coding for PBP3) of Haemophilus influenzae with aminopenicillin resistance and to evaluate the 2017 European Committee for Antibiotic Susceptibility Testing (EUCAST) guidelines for clinical categorization of ampicillin, amoxicillin, and amoxicillin-clavulanate for strains with mutated PBP3 conferring resistance (rPBP3). A panel of 91 H. influenzae isolates was genetically characterized by sequencing of the fstI gene. For all the studied isolates, a screening with benzylpenicillin 1U (BP1) was carried out and minimum inhibitory concentrations (MICs) of ampicillin, amoxicillin, and amoxicillin-clavulanate were tested and interpreted according to EUCAST recommendations. ftsI sequence analysis revealed a total of 14 different amino acid substitutions in PBP3. The substitution patterns most commonly observed were [D350N, M377I, A502V, N526K] among the bla-positive rPBP3 strains (37.5%) and [D350N, A502T, N526K] among the bla-negative rPBP3 strains (24.5%). Screening with BP1 was able to correctly categorize 100% of the bla-negative sPBP3 strains, 100% of the bla-positive strains, and 92% of the bla-negative rPBP3 ones. Only 29% of the bla-negative rPBP3 strains evaluated displayed ampicillin MICs above the current EUCAST resistant breakpoint defined at 1 μg/ml. The PBP3 substitution patterns of the strains evaluated are similar to the ones observed in previous Spanish and European studies. Although the screening with BP1 proved to be adequate in the detection of bla-negative rPBP3 strains, these cannot be reliably identified by current 2018 EUCAST breakpoints for ampicillin.

Keywords

Haemophilus influenzae β-lactam resistance ftsI rPBP3 EUCAST 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

For this type of study, formal consent is not required.

Informed consent

Informed consent was obtained from all individual participants included in the study.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Service of MicrobiologyUniversity Hospital Marqués de Valdecilla-IDIVALSantanderSpain
  2. 2.Service of MicrobiologyUniversity Hospital Marqués de Valdecilla-IFIMAVSantanderSpain
  3. 3.Department of Molecular BiologyUniversity of CantabriaSantanderSpain
  4. 4.Unit of MicrobiologyHospital Universitario Reina SofíaCórdobaSpain
  5. 5.Department of MicrobiologyUniversity of CórdobaCórdobaSpain
  6. 6.Instituto Maimónides de Investigación Biomédica de Córdoba (IMIBIC)CórdobaSpain

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