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- Inverse PCR for subtyping of Acinetobacter baumannii carrying ISAba1
- Shukho Kim , Yun-Ju Park , Jungmin Kim
- J. Microbiol. 2016;54(5):376-380. Published online April 20, 2016
- DOI: https://doi.org/10.1007/s12275-016-6038-3
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- 3 Crossref
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Abstract
- Acinetobacter baumannii has been prevalent in nosocomial infections, often causing outbreaks in intensive care units. ISAba1 is an insertion sequence that has been identified only in A. baumannii and its copy number varies among strains. It has been reported that ISAba1 provides a promoter for blaOXA-51-like, blaOXA-23-like, and blaampC, which are associated with the resistance of A. baumannii to carbapenems and cephalosporins. The main purpose of this study was to develop a novel inverse PCR method capable of typing A. baumannii strains. The method involves three major steps: cutting of genomic DNA with a restriction enzyme, ligation, and PCR. In the first step, bacterial genomic DNA was digested with DpnI. In the second step, the digested genomic DNAs were ligated to form intramolecular circular DNAs. In the last step, the ligated circular DNAs were amplified by PCR with primers specific for ISAba1 and the amplified PCR products were electrophoresed. Twenty-two clinical isolates of A. baumannii were used for the evaluation of the inverse PCR (iPCR) typing method. Dendrogram analysis revealed two major clusters, similar to pulsed-field gel electrophoresis (PFGE) results. Three ISAba1-associated genes – blaampC, blaOXA-66-like, and csuD – were amplified and detected in the clinical isolates. This novel iPCR typing method is comparable to PFGE in its ability to discriminate A. baumannii strains, and is a promising molecular epidemiological tool for investigating A. baumannii carrying ISAba1.
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Citations
Citations to this article as recorded by
- DNA sonication inverse PCR for genome scale analysis of uncharacterized flanking sequences
David E. Alquezar‐Planas, Ulrike Löber, Pin Cui, Claudia Quedenau, Wei Chen, Alex D. Greenwood, Susan Johnston
Methods in Ecology and Evolution.2021; 12(1): 182. CrossRef -
Update on the Epidemiological Typing Methods for
Acinetobacter Baumannii
Rayane Rafei, Marwan Osman, Fouad Dabboussi, Monzer Hamze
Future Microbiology.2019; 14(12): 1065. CrossRef - Identification and characterization of a novel cold-tolerant extracellular protease from Planococcus sp. CGMCC 8088
Kun Chen, Qingshan Mo, Huan Liu, Feiyan Yuan, Haonan Chai, Fuping Lu, Huitu Zhang
Extremophiles.2018; 22(3): 473. CrossRef
- DNA sonication inverse PCR for genome scale analysis of uncharacterized flanking sequences
Research Support, Non-U.S. Gov't
- Cloning and Characterization of a Na+/H+ Antiporter Gene of the Moderately Halophilic Bacterium Halobacillus aidingensis AD-6T
- Ya Jie Zou , Li Fu Yang , Lei Wang , Su Sheng Yang
- J. Microbiol. 2008;46(4):415-421. Published online August 31, 2008
- DOI: https://doi.org/10.1007/s12275-008-0009-2
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- 14 Scopus
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Abstract
- A gene encoding a Na+/H+ antiporter was obtained from the genome of Halobacillus aidingensis AD-6T, which was sequenced and designated as nhaH. The deduced amino acid sequence of the gene was 91% identical to the NhaH of H. dabanensis, and shared 54% identity with the NhaG of Bacillus subtilis. The cloned gene enable the Escherichia coli KNabc cell, which lack all of the major Na+/H+ antiporters, to grow in medium containing 0.2 M NaCl or 10 mM LiCl. The nhaH gene was predicted to encode a 43.5 kDa protein (403 amino acid residues) with 11 putative transmembrane regions. Everted membrane vesicles prepared from E. coli KNabc cells carrying NhaH exhibited Na+/H+ as well as Li+/H+ antiporter activity, which was pH-dependent with the highest activity at pH 8.0, and no K+ /H+ antiporter activity was detected. The deletion of hydrophilic C-terminal amino acid residues showed that the short C-terminal tail was vital for Na+/H+ antiporter activity.
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