Annals of Laboratory Medicine

Korean Society for Laboratory Medicine (대한진단검사의학회)
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Rapid Detection of Pseudomonas aeruginosa and Acinetobacter baumannii Harboring blaVIM-2, blaIMP-1 and blaOXA-23 Genes by Using Loop-Mediated Isothermal Amplification Methods

  • Kim, Hye Jin (Department of Microbiology and Immunology, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine) ;
  • Kim, Hyung Sun (Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine) ;
  • Lee, Jae Myun (Department of Microbiology and Immunology, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine) ;
  • Yoon, Sang Sun (Department of Microbiology and Immunology, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine) ;
  • Yong, Dongeun (Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine)
  • Received : 2015.05.21
  • Accepted : 2015.10.13
  • Published : 2016.01.01
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Abstract

Background: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) and Acinetobacter baumannii (CRAB) are the leading causes of nosocomial infections. A rapid and sensitive test to detect CRPA and CRAB is required for appropriate antibiotic treatment. We optimized a loop-mediated isothermal amplification (LAMP) assay to detect the presence of $bla_{VIM-2}$, $bla_{IMP-1}$, and $bla_{OXA-23}$, which are critical components for carbapenem resistance. Methods: Two sets of primers, inner and outer primers, were manually designed as previously described. The LAMP buffer was optimized (at 2mM $MgSO_4$) by testing different concentrations of $MgSO_4$. The optimal reaction temperature and incubation time were determined by using a gradient thermocycler. Then, the optimized $bla_{VIM-2}$, $bla_{IMP-1}$, and $bla_{OXA-23}$ LAMP reactions were evaluated by using 120 P. aeruginosa and 99 A. baumannii clinical isolates. Results: Only one strain of the 100 CRPA isolates harbored $bla_{IMP-1}$, whereas none of them harbored $bla_{VIM-2}$. These results indicate that the acquisition of $bla_{VIM-2}$ or $bla_{IMP-1}$ may not play a major role in carbapenem resistance in Korea. Fifty two strains of the 75 CRAB isolates contained $bla_{OXA-23}$, but none contained $bla_{VIM-2}$ and $bla_{IMP-1}$ alleles. Conclusions: Our results demonstrate the usefulness of LAMP for the diagnosis of CRPA and CRAB.

Keywords

Acknowledgement

Supported by : Ministry for Health, Welfare, and Family Affairs

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