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Rapid detection of Pseudomonas aeruginosa and Acinetobacter baumannii Harboring bla(VIM-2), bla(IMP-1) and bla(OXA-23) genes by using loop-mediated isothermal amplification methods.

 Hye Jin Kim  ;  Hyung Sun Kim  ;  Jae Myun Lee  ;  Sang Sun Yoon  ;  Dongeun Yong 
 Annals of Laboratory Medicine, Vol.36(1) : 15-22, 2016 
Journal Title
 Annals of Laboratory Medicine 
Issue Date
Acinetobacter baumannii/genetics ; Acinetobacter baumannii/isolation & purification* ; Anti-Bacterial Agents/pharmacology* ; Carbapenems/pharmacology* ; Drug Resistance, Bacterial/genetics* ; Genes, Bacterial* ; Nucleic Acid Amplification Techniques ; Pseudomonas aeruginosa/genetics ; Pseudomonas aeruginosa/isolation & purification* ; Sensitivity and Specificity
Acinetobacter baumannii ; Carbapenem resistance ; Loop-mediated isothermal amplification (LAMP) ; Pseudomonas aeruginosa ; bla(IMP-1) ; bla(OXA-23) ; bla(VIM-2)
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 MgSO₄) by testing different concentrations of MgSO₄. 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.
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1. College of Medicine (의과대학) > Dept. of Laboratory Medicine (진단검사의학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Microbiology (미생물학교실) > 1. Journal Papers
Yonsei Authors
용동은(Yong, Dong Eun) ORCID logo https://orcid.org/0000-0002-1225-8477
윤상선(Yoon, Sang Sun) ORCID logo https://orcid.org/0000-0003-2979-365X
이재면(Lee, Jae Myun) ORCID logo https://orcid.org/0000-0002-5273-3113
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