Rapid Detection of Pseudomonas aeruginosa and Acinetobacter baumannii Harboring blaVIM-2, blaIMP-1 and blaOXA-23 Genes by Using Loop-Mediated Isothermal Amplification Methods.
- Author:
Hye Jin KIM
1
;
Hyung Sun KIM
;
Jae Myun LEE
;
Sang Sun YOON
;
Dongeun YONG
Author Information
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords: Loop-mediated isothermal amplification (LAMP); Carbapenem resistance; blaVIM-2; blaIMP-1; blaOXA-23; Pseudomonas aeruginosa; Acinetobacter baumannii
- MeSH: Acinetobacter baumannii/genetics/*isolation & purification; Anti-Bacterial Agents/*pharmacology; Carbapenems/*pharmacology; Drug Resistance, Bacterial/*genetics; *Genes, Bacterial; Nucleic Acid Amplification Techniques; Pseudomonas aeruginosa/genetics/*isolation & purification; Sensitivity and Specificity
- From:Annals of Laboratory Medicine 2016;36(1):15-22
- CountryRepublic of Korea
- Language:English
- 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 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.
