Yonsei Med J.  2015 Jul;56(4):928-934. 10.3349/ymj.2015.56.4.928.

In Vivo Selection of Pan-Drug Resistant Acinetobacter baumannii during Antibiotic Treatment

• Kim Y
• Bae IK
• Jeong SH
• Yong D
• Lee K

• KMID: 2366331
• DOI: http://doi.org/10.3349/ymj.2015.56.4.928

Abstract

PURPOSE
Colistin resistance in Acinetobacter baumannii (A. baumannii) is mediated by a complete loss of lipopolysaccharide production via mutations in lpxA, lpxC, and lpxD gene or lipid A modifications via mutations in the pmrA and pmrB genes. However, the exact mechanism of therapy-induced colistin resistance in A. baumannii is not well understood.
MATERIALS AND METHODS
We investigated the genotypic and phenotypic changes that underlie pan-drug resistance mechanisms by determining differences between the alterations in extensively drug-resistant (XDR) A. baumannii (AB001 and AB002) isolates and a pan-drug resistant (PDR) counterpart (AB003) recovered from one patient before and after antibiotic treatment, respectively.
RESULTS
All three clinical isolates shared an identical sequence type (ST138), belonging to the global epidemic clone, clonal complex 92, and all produced OXA-23 carbapenemase. The PDR AB003 showed two genetic differences, acquisition of armA gene and an amino acid substitution (Glu229Asp) in pmrB gene, relative to XDR isolates. No mutations were detected in the pmrA, pmrC, lpxA, lpxC, or lpxD genes in all three isolates. In matrix-assisted laser desorption ionization-time of flight analysis, the three isolates commonly showed two major peaks at 1728 m/z and 1912 m/z, but peaks at 2034 m/z, 2157 m/z, 2261 m/z, and 2384 m/z were detected only in the PDR A. baumannii AB003 isolate.
CONCLUSION
Our results show that changes in lipid A structure via a mutation in the pmrB gene and acquisition of armA gene might confer resistance to colistin and aminoglycosides to XDR A. baumannii strains, resulting in appearance of a PDR A. baumannii strain of ST138.

Keyword

pmrB gene; armA gene; colistin; lipid A; Acinetobacter baumannii

MeSH Terms

Acinetobacter Infections/*drug therapy/microbiology
Acinetobacter baumannii/*drug effects/*genetics/isolation & purification
Aged
Anti-Bacterial Agents/*pharmacology/therapeutic use
Bacterial Proteins/*genetics
Colistin/*pharmacology/therapeutic use
*Drug Resistance, Bacterial
Electrophoresis, Gel, Pulsed-Field
Genotype
Humans
Male
Microbial Sensitivity Tests
Molecular Typing
Mutation
Polymerase Chain Reaction
Transcription Factors
beta-Lactamases
Anti-Bacterial Agents
Bacterial Proteins
Colistin
Transcription Factors
beta-Lactamases

Figure

• 

  Fig. 1 SmaI-macrorestiction patterns of two colistin-susceptible isolates (AB001 and AB002) and a colistin-resistant isolate (AB003). All three isolates showed 100% of similarity.

• 

  Fig. 2 Mass spectrometry of lipid A extracted from the colistin-susceptible isolates (A) AB001 and (B) AB002 and their colistin-resistant counterpart (C) AB003 with Glu229Asp in pmrB gene. In MALDI-TOF analysis, AB001, AB002, and AB003 isolates all showed two major peaks at 1728 m/z and 1912 m/z, but peaks at 2034 m/z, 2157 m/z, 2261 m/z, and 2384 m/z were detected only in the PDR A. baumannii AB003 isolate. MALDI-TOF, matrix-assisted laser desorption ionization-time of flight; PDR, pan-drug resistant.

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