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Isolation and Identification of Waterborne Antibiotic-Resistant Bacteria and Molecular Characterization of their Antibiotic Resistance Genes
Published on: March 3, 2023
Genetic characterization and diversity of Streptococcus agalactiae isolates with macrolide resistance
Monika Brzychczy-Włoch1, Tomasz Gosiewski1, Małgorzata Bodaszewska1
1Chair of Microbiology, Jagiellonian University Medical College, Krakow, Poland.
Abstract:
Macrolide resistance in 169 Streptococcus agalactiae [group B streptococcus (GBS)] isolates originating from pregnant carriers was investigated. Using multiplex PCR the presence of genes encoding erythromycin resistance and capsular polysaccharides, as well as surface proteins, was determined. Random amplification of polymorphic DNA (RAPD) and PFGE were used to characterize specific clones among the isolates. In the examined population of women, erythromycin-resistant strains were found in 4.5 % of patients, whereas clindamycin-resistant strains were found in 3 % of patients, which was 16 % of strains resistant to erythromycin and 10 % of strains resistant to clindamycin among GBS isolates, respectively. Among the isolates, the largest percentage was represented by the constitutive macrolide-lincosamide-streptogramin B (cMLS(B)) phenotype (63 %), then the inductive macrolide-lincosamide-streptogramin B (iMLS(B)) phenotype (26 %) and the macrolide resistance (M) phenotype (11 %). The ermB gene was indicated in all isolates with the cMLS(B) phenotype and V serotype, whereas mefA/mefE genes were found in isolates with the M phenotype and Ia serotype. Among resistance isolates, serotype V was predominant (67 %), followed by serotypes II (15 %), Ia (11 %) and III (7 %). The most common surface protein encoding genes were alp3 (70 %), then rib (11 %), epsilon (7.5 %), bca (7.5 %) and alp2 (4 %). A statistically significant relationship between macrolide resistance, serotype V and the alp3 gene was demonstrated. PFGE, in comparison to the RAPD method, gave better genetic discrimination of GBS isolates. A relatively high genetic diversity among investigated strains was shown. In addition, the largest genetic homogeneity was found in serotype V.
Insights
Macrolide resistance in pregnant women
Area of Science:
- Microbiology
- Genetics
- Epidemiology
Background:
- Group B Streptococcus (GBS) is a significant pathogen, particularly in pregnant women.
- Macrolide resistance in GBS is a growing public health concern.
- Understanding resistance mechanisms and genetic diversity is crucial for effective treatment.
Purpose of the Study:
- To investigate macrolide resistance in Streptococcus agalactiae (GBS) isolates from pregnant carriers.
- To determine the prevalence of resistance genes, serotypes, and surface proteins.
- To analyze the genetic diversity and relationships between resistance and specific GBS characteristics.
Main Methods:
- Multiplex PCR was used to detect resistance genes (ermB, mefA/mefE) and genes for capsular polysaccharides and surface proteins.
- Random Amplification of Polymorphic DNA (RAPD) and Pulse Field Gel Electrophoresis (PFGE) were employed for clonal characterization.
- Phenotypic characterization of macrolide resistance (cMLS(B), iMLS(B), M) was performed.
Main Results:
- Macrolide resistance was observed in 4.5% of pregnant carriers, with clindamycin resistance in 3%.
- The constitutive macrolide-lincosamide-streptogramin B (cMLS(B)) phenotype was most common (63%), followed by iMLS(B) (26%) and M (11%).
- Serotype V and the alp3 surface protein gene were significantly associated with macrolide resistance. PFGE provided better genetic discrimination than RAPD.
Conclusions:
- A significant prevalence of macrolide resistance exists in GBS isolates from pregnant women.
- Serotype V and the alp3 gene are key factors associated with macrolide resistance in this population.
- PFGE is a superior method for assessing genetic diversity in GBS isolates, revealing high diversity overall but homogeneity within serotype V.
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