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Rapid and Specific Detection of Acinetobacter baumannii Infections Using a Recombinase Polymerase Amplification/Cas12a-based System
Published on: April 25, 2025
Whole-Genome Sequences of Five Acinetobacter baumannii Strains From a Child With Leukemia M2
Jetsi Mancilla-Rojano1,2, Semiramis Castro-Jaimes3, Sara A Ochoa1
1Laboratorio de Investigación en Bacteriología Intestinal, Hospital Infantil de México Federico Gómez, Mexico City, Mexico.
Abstract:
Acinetobacter baumannii is an opportunistic pathogen and is one of the primary etiological agents of healthcare-associated infections (HAIs). A. baumannii infections are difficult to treat due to the intrinsic and acquired antibiotic resistance of strains of this bacterium, which frequently limits therapeutic options. In this study, five A. baumannii strains (810CP, 433H, 434H, 483H, and A-2), all of which were isolated from a child with leukemia M2, were characterized through antibiotic susceptibility profiling, the detection of genes encoding carbapenem hydrolyzing oxacillinases, pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), adherence and invasion assays toward the A549 cell line, and the whole-genome sequence (WGS). The five strains showed Multidrug resistant (MDR) profiles and amplification of the bla OXA-23 gene, belonging to ST758 and grouped into two PFGE clusters. WGS of 810CP revealed the presence of a circular chromosome and two small plasmids, pAba810CPa and pAba810CPb. Both plasmids carried genes encoding the Sp1TA system, although resistance genes were not identified. A gene-by-gene comparison analysis was performed among the A. baumannii strains isolated in this study and others A. baumannii ST758 strains (HIMFG and INCan), showing that 86% of genes were present in all analyzed strains. Interestingly, the 433H, 434H, and 483H strains varied by 8-10 single-nucleotide variants (SNVs), while the A2 and 810CP strains varied by 46 SNVs. Subsequently, an analysis using BacWGSTdb showed that all of our strains had the same resistance genes and were ST758. However, some variations were observed in relation to virulence genes, mainly in the 810CP strain. The genes involved in the synthesis of hepta-acylated lipooligosaccharides, the pgaABCD locus encoding poly-β-1-6-N-acetylglucosamine, the ompA gene, Csu pili, bap, the two-component system bfms/bfmR, a member of the phospholipase D family, and two iron-uptake systems were identified in our A. baumannii strains genome. The five A. baumannii strains isolated from the child were genetically different and showed important characteristics that promote survival in a hospital environment. The elucidation of their genomic sequences provides important information for understanding their epidemiology, antibiotic resistance, and putative virulence factors.
Insights
This study characterized five multidrug-resistant Acinetobacter baumannii strains from a leukemia patient. Whole-genome sequencing revealed genetic differences and potential virulence factors, aiding understanding of Acinetobacter baumannii epidemiology and resistance.
Area of Science:
- Microbiology
- Genomics
- Infectious Diseases
Background:
- Acinetobacter baumannii is a significant cause of healthcare-associated infections.
- Antibiotic resistance in A. baumannii complicates treatment options.
- This study focuses on strains isolated from a pediatric leukemia patient.
Purpose of the Study:
- To comprehensively characterize five A. baumannii strains.
- To investigate antibiotic resistance profiles and genetic relatedness.
- To identify potential virulence factors through whole-genome sequencing.
Main Methods:
- Antibiotic susceptibility profiling
- Detection of carbapenemase genes (blaOXA-23)
- Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST)
- Whole-genome sequencing (WGS) of strain 810CP
- Adherence and invasion assays
Main Results:
- All five strains exhibited multidrug-resistant (MDR) profiles and carried the blaOXA-23 gene.
- Strains belonged to sequence type 758 (ST758) and formed two PFGE clusters.
- WGS revealed genetic variations among strains, with notable differences in virulence genes, particularly in strain 810CP.
- Genes associated with lipooligosaccharide synthesis, biofilm formation (pgaABCD), outer membrane proteins (ompA), pili (Csu), and iron uptake were identified.
Conclusions:
- The five A. baumannii strains, despite sharing ST758 and resistance genes, displayed genetic diversity.
- These strains possess characteristics conducive to survival in healthcare settings.
- Genomic insights are crucial for understanding A. baumannii epidemiology, antibiotic resistance, and virulence.
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