Genotypic Investigation of Multidrug-Resistant Pseudomonas aeruginosa from Clinical Isolates in Korea, 2010

2010년도 국내 임상에서 분리한 다제내성 녹농균의 유전자형 조사

  • Received : 2012.11.02
  • Accepted : 2012.12.14
  • Published : 2012.12.31


Pseudomonas aeruginosa is an opportunistic Gram-negative bacterium that causes serious infection, particularly in immunocompromised patients. Also, P. aeruginosa possessing carbapenem-resistant metallo-${\beta}$-lactamases (MBL) has been reported with increasing frequency in Korea. We therefore analyzed the level of multidrug-resistant clinical P. aeruginosa isolated from a secondary hospital in Korea in 2010. A total of 92 isolates of P. aeruginosa were collected from Sahmyook Medical Center in 2010. Susceptibility to antimicrobial agents was determined by analysis of the minimum inhibitory concentration test; the inhibitor-potentiated disk diffusion (IPD) test was performed for MBL detection. RAPD-PCR was used for genotyping to rapidly characterize P. aeruginosa strains isolated from clinical patients. The percentages of non-susceptible isolates were as follows: 40.2% to ceftazidime, 58.7% to meropenem, 56.5% to gentamicin, 46.7% to tobramycin, 62.0% to ciprofloxacin and 97.8% to chloramphenicol. The 29 multidrug-resistant strains were screened by the IPD test: of the 21 PCR-positive isolates, 19 were IPM-1 producers and 2 were VIM-2 producers. Among the 19 IMP-1-producing P. aeruginosa isolates, 16 isolates showed similar patterns, and three different banding patterns were observed. The proportion of IMP-1-producing multidrug-resistant P. aeruginosa from clinical isolates steadily increased in this secondary hospital in Korea in 2010. This study provides information about the antimicrobial-resistant patterns and genotype of multidrug-resistant P. aeruginosa isolated from clinical isolates in Korea, 2010.


Pseudomonas aeruginosa;antimicrobial resistance;metallo-${\beta}$-lactamases (MBL);multidrug-resistant


Supported by : Korea Small and Medium Business Administration, Sahmyook University


  1. Anna, S.L., Antonio, A.B., Juliana, P., Marcio, V.S., Ivan, S.M., Erico, G.A., Edison, I.M., and Silvia, F.C. 1999. Intravenous colistin as therapy for nosocomial infections caused by multidrug-resistant Pseudomonas aeruginosa and Acinetobacter baumannii. Clin. Infect. Dis. 28, 1008-1011.
  2. Bush, K. 1998. Metallo-${\beta}$-lactamases: a class apart. Clin. Infect. Dis. 27, 48-53.
  3. Chu, Y.-W., Afzal-Shah, M., Houang, E.T., Palepou, M.I., Lyon, D.J., Woodford, N., and Livermore, D. 2001. IMP-4, a novel metallo-${\beta}$ -lactamase from nosocomial Acinetobacter spp. collected in Hong Kong between 1994 and 1998. Antimicrob. Agents Chemother. 45, 710-714.
  4. Clinical and Laboratory Standards Institute (CLSI). 2007. Performance standards for antimicrobial susceptibility testing; seventh informational supplement 27. Suit, USA, 98-114.
  5. Fernandez-Cuenca, F., Martinez-Martinez, L., Conejo, M.C., Ayala, J.A., Perea, E.J., and Pascual, A. 2003. Relationship between ${\beta}$-lactamase production, outer membrane protein and penicillinbinding protein profiles on the activity of carbapenems against clinical isolates of Acinetobacter baumannii. J. Antimicrob. Chemother. 51, 565-574.
  6. Gibb, A.P., Tribuddharat, C., Moore, R.A., Louie, T.J., Krulicki, W., Livermore, D.M., Palepou, M.F., and Woodford, N. 2002. Nosocomial outbreak of carbapenem-resistant Pseudomonas aeruginosa with a new blaIMP allele, $blaI_{MP-7}$. Antimicrob. Agents Chemother. 46, 255- 258.
  7. Iyobe, S., Kusadokoro, H., Ozaki, J., Matsumura, N., Minami, S., Haruta, S., Sawai, T., and O'Hara, K. 2000. Amino acid substitutions in a variant of IMP-1 metallo-${\beta}$-lactamase. Antimicrob. Agents Chemother. 44, 2023-2027.
  8. Jeon, B.C., Jeong, S.H., Bae, I.K., Kwon, S.B., Lee, K., Young, D., Lee, J.H., Song, J.S., and Lee, S.H. 2005. Investigation of a nosocomial outbreak of imipenem-resistant Acinetobacter baumannii producing the OXA-23 beta-lactamase in Korea. J. Clin. Microbiol. 43, 2241- 2245.
  9. Kim, J.H., Jeong, S.H., Kim, B.N., Jeong, T.J., Kim, T.S., Kim, J.C., and Jeon, B.C. 2002. Genotypes of metallo-${\beta}$-lactamase produced by imipenem-resistant Acinetobacter spp. isolates from clinical specimens. Korean J. Infect. Dis. 34, 360-366.
  10. Kim, J.R., Lee, D.K., An, H.M., Kim, M.J., Lee, S.W., Cha, M.K., Lee, K.O., and Ha, N.J. 2011. Antimicrobial activity of commonly used antibiotics and DNA fingerprint analysis of Pseudomonas aeruginosa obtained from clinical isolates and unchlorinated drinking water in Korea, 2010. Arch. Pharm. Res. 34, 1353-1361.
  11. Kim, I.S., Oh, W.I., Song, J.H., and Lee, N.Y. 2004. Screening and identification of metallo-${\beta}$-lactamase gene in clinical isolates of imipenem-resistant Pseudomonas aeruginosa. Korean J. Lab. Med. 24, 177-182.
  12. Koeuth, T., Versalovic, J., and Lupski, J.R. 1995. Differential subsequence conservation of interspersed repetitive Streptococcus pneumoniae BOX elements in diverse bacteria. Genome Res. 5, 408- 418.
  13. Landman, D. and Quale, J. 2002. Pseudomonas aeruginosa and Acinetobacter baumannii: pathogens of the millennium. Ochsner Clinic Reports on Serious Hospital Infection. 14, 1-7.
  14. Lee, K., Ha, G.Y., Shin, B.M., Kim, J.J., Kang, J.O., Jang, S.J., Yong, D., Chong, Y., and Korean Nationwide Surveillance of Antimicrobial Resistance (KONSAR) group. 2004. Metallo-${\beta}$-lactamase-producing Gram-negative bacilli in Korean nationwide surveillance of antimicrobial resistance group hospitals in 2003: continued prevalence of VIM-producing Pseudomonas spp. and increase of IMP producing Acinetobacter spp. Diagn. Microbiol. Infect. Dis. 50, 51-58.
  15. Lee, K., Lim, J.B., Yum, J.H., Yong, D., Chong, Y., Kim, J.M., and Livermore, D.M. 2002. blaVIM-2 Cassette-containing novel integrons in metallo-${\beta}$-lactamase-producing Pseudomonas aeruginosa and Pseudomonas putida isolates disseminated in a Korean hospital. Antimicrob. Agents Chemother. 46, 1053-1058.
  16. Lee, K.W., Park, A.J., Moon, Y.K., Lee, H.J., Cho, J.H., Kang, J.O., Yong, D.G., Chong, Y.S., and KONSAR group. 2009. Metallo-${\beta}$-lactamase-producing Pseudomonas spp. in Korea: High prevalence of isolates with VIM-2 type and rmergence of isolates with IMP-1 type. Yonsei Med. J. 50, 335-339.
  17. Levasque, C., Piche, L., Larose, C., and Roy, P.H. 1995. PCR mapping of integrons reveals several novel combinations of resistance genes. Antimicrob. Agents Chemother. 39, 185-191.
  18. Livermore, D.M. 2002. Multiple mechanisms of antimicrobial resistance in Pseudomonas aeruginosa: our worst nightmare? Clin. Infect. Dis. 34, 634-640.
  19. Livermore, D.M. and Woodford, N. 2000. Carbapenemases: a problem in waiting? Curr. Opin. Microbiol. 3, 489-495.
  20. Oh, E.J., Lee, S., Park, Y.J., Park, J.J., Park, K., Kim, S.I., Kang, M.W., and Kim, B.K. 2003. Prevalence of metallo-beta-lactamase among Pseudomonas aeruginosa and Acinetobacter baumannii in a Korean university hospital and comparison of screening methods for detecting metallo-beta-lactamase. J. Microbiol. Methods 54, 411- 418.
  21. Riccio, M.L., Franceschini, N., Boschi, L., Caravelli, B., Cornaglia, G., Fontana, R., Amicosante, G., and Rossolini, G.M. 2000. Characterization of the metallo-${\beta}$-lactamase determinant of Acinetobacter baumannii AC-54/97 reveals the existence of $bla_{IMP}$ allelic variants carried by gene cassettes of different phylogeny. Antimicrob. Agents Chemother. 44, 1229-1235.
  22. Senda, K., Arakawa, Y., Nakashima, K., Ito, H., Ichiyama, S., Shimokata, K., Kato, N., and Ohta, M. 1996. Multifocal outbreaks of metallobeta- lactamase-producing Pseudomonas aeruginosa resistant to broad-spectrum beta-lactams, including carbapenems. Antimicrob. Agents Chemother. 40, 349-353.
  23. Tam, V.H., Chang, K.T., LaRocco, M.T., Schilling, A.N., McCauley, S.K., Poole, K., and Garey, K.W. 2007. Prevalence, mechanisms, and risk factors of carbapenem resistance in bloodstream isolates of Pseudomonas aeruginosa. Diagn. Microbiol. 58, 309-314.
  24. Walsh, T.R., Toleman, M.A., Poirel, L., and Nordmann, P. 2005. Metallo- ${\beta}$-lactamases: the quiet before the storm? Clin. Microbiol. 18, 306-325.
  25. Yong, D.G. 2009. Metallo-${\beta}$-lactamase producing Gram-negative Bacilli. Korean J. Clin. Microbiol. 12, 103-109.
  26. Yong, D., Lee, K., Yum, J.H., Shin, H.B., Rossolini, G.M., and Chong, Y. 2002. Imipenem-EDTA disk method for differentiation of metallo-beta-lactamase-producing clinical isolates of Pseudomonas spp. and Acinetobacter spp. J. Clin. Microbiol. 40, 3798-3801.
  27. Zhiyong, Z., Xiaoju, L., and Yanyu, G. 2003. Metallo-${\beta}$-lactamases of non-fermenting Gram-negative bacteria. Rev. Med. Microbiol. 14, 79 -93.