The antibiotic resistance, gelatinase production and biofilm formation among Enterococcus strains &ndash; the correlation analysis using PCR techniques

Paulina Pecyna; Marcelina Jaworska; Monika Rosochowicz; Julia Ostapowicz; Julia Lipowicz; Marianna Karwacka; Paulina Wiacek; Oliwia Szymanowicz; Marzena Gajecka

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The antibiotic resistance, gelatinase production and biofilm formation among Enterococcus strains – the correlation analysis using PCR techniques

Paulina Pecyna

Marcelina Maria Jaworska

^*,

Paulina Wiacek

Oliwia Szymanowicz

Marzena Gajecka

¹ Department of Genetisc and Pharmaceutical Microbiology

Published: 26 April 2021 by MDPI in The 1st International Electronic Conference on Antibiotics session Poster

https://doi.org/10.3390/ECA2021-09642 (registering DOI)

Abstract:

Background: Enterococcus spp. are among the most frequently detected Gram-positive bacteria in the intestinal flora. These bacteria have developed many defense mechanisms that allow to survive in unfavorable conditions. In many cases Enterococcus spp. are resistant to commonly used antibiotics – vancomycin and teicoplanin (VRE - Vancomycin-Resistant Enterococcus) or gentamicin (HLAR -High-Level Aminoglycoside Resistance)and produce enzymes like gelatinase (gelE gene). An observed significant increase in the number of VRE-positive isolates and the ability to form a biofilm influence the spread of multi-drug resistant strains.

Aim: This study aimed to analyze the presence of vanA and vanB genes, which are responsible for the resistance to vancomycin and teicoplanin. Identification of genes responsible for the formation of biofilm (esp, fsrA, fsrB) and the production of the gelatinase enzyme (gelE) were also assumed.

Materials and Methods: Enterococcus bacteria isolated on D-Cocogel medium were collected from 56 hospital patients’rectal swabs. DNA from colonies of each strain was isolated using a heat-shock method in the TE buffer. The PCR reactions were conducted using specific primers to each of the evaluated genes: vanA, vanB, gelE, gelEfsr, esp, fsrA, fsrB.

Results: Vancomycin resistance was confirmed among 93% of tested Enterococcus strains. The vanA and 18% vanB genes were identified in 75% of the strains. The 12% of Enterococcus strains possessed the ability to produce gelatinase, among which 5% and 2% contained gelE gene, and gene gelEfsr, respectively. The remaining 5% of the strains have both genes gelEfsr and gelE. The 34% of tested bacteria strains showed the presence of analyzed genes responsible for biofilm production (gene esp in 58 % Enterococcus biofilm productive strains, and fsrA and fsrB genes among 42% strains). In all (seven) gelatinase production strains there was a parallel presence of genes fsrA and fsrB (responsible for biofilm formation).