Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
권호 표지

Prevalence of Extended Spectrum $\beta-Lactamase-Producing$ Clinical Isolates of Escher­ichia coli in a University Hospital, Korea

국내 대학병원에서 분리된 Eschepichia coli의 Extended-spectrum $\beta-Lactamase$ (ESBL) 현황

  • Lee Kyenam (Department of Biology and Culture Collection of Antimicrobial Resistant Microbes, Seoul Women's University) ;
  • Kim Woo-Joo (Department of Infectious Diseases, Kuro Hospital, Korea University) ;
  • Lee Yeonhee (Department of Biology and Culture Collection of Antimicrobial Resistant Microbes, Seoul Women's University)
  • 이계남 (서울여자대학교 생물학과 항생제내성균주은행) ;
  • 김우주 (고려대학교 의과대학 감염내과) ;
  • 이연희 (서울여자대학교 생물학과 항생제내성균주은행)
  • Published : 2004.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

Recently, the rapid increase and global spread of extended-spectrum $\beta-lactamase$ producing clinical isolates has become a serious problem. The incidence of extended-spectrum $\beta-lactamase$ producing clinical isolates of Escherichia coli in Korea and susceptibility to antimicrobial agents were investigated. Total 233 isolates of E. coli were obtained from urine from hospitalized patients in Guro hospital, Korea University in 2001. One hun­dred and eighty four isolates $(78.9\%)$ were resistant to ampicillin, 80 isolates $(34.3\%)$ were resistant to ceph­alothin, 93 isolates $(39.9\%)$ were resistant to gentamicin, and 64 isolates $(27.5\%)$ were resistant to norfloxacin. Among 233 isolates, 17 isolates $(7.3\%)$ were positive as determined by the double disk synergy test. When min­imal inhibitory concentrations were assayed with additional 6 antimicrobial agents, 13 isolates $(76.5\%)$ were multi-drug resistant to at least four different class antimicrobial agents. Extended-spectrum $\beta-lactamase$ were characterized with isoelectric focusing gel electrophoresis and DNA sequencing. They were TEM-1 in 5 iso­lates, TEM-15 in 1 isolate, TEM-20 in 1 isolate, TEM-52 in 4 isolates, TEM-1 and AmpC in 2 isolates, TEM-1 and OXA-30 in 1 isolate, TEM-1 and OXA-33 in 1 isolate, TEM-1, CTX-M-3, and AmpC in 1 isolate, but SHV was not detected. Antimicrobial resistance genes were transferred to animal isolate of E. coli (CCARM No. 1203) by the filter mating method. Extended spectrum $\beta-lactamase$ producers studied in the current study have low correlation to each other as determined by random amplified polymorphic DNA and pulsed field gel elec­trophoresis. This is a contradictory result from the general hypothesis that extended-spectrum $\beta-lactamase$ pro­ducers in one hospital is a result from a clonal spread.

최근 extended-spectrum $\beta-lactamase$ 생산 임상 균주의 급속한 증가와 확산은 매우 심각한 문제를 야기하고 있다. 이에 국내에서 extended-spectrum $\beta-lactamase$ 생산 임상 균주의 발생율을 파악하고자 국내 한 대학병원에 입원한 환자로부터 대장균을 분리하여 항생제 감수성 검사를 실시하였다. 충 233균주 중 184균주 $(78.9\%)$가 ampicillin에 대해 내성을 나타냈으며, 80균주$(34.3\%)$가 cephalothin에, 93균주$(39.9\%)$가 gentamicin에, 64균주$(27.5\%)$가 norfloxacin에 대해 내성을 나타내었다. 이중 17균주$(7.3\%)$가 double disk synergy test에 의해 양성반응을 나타낸 것으로 확인되었고, 이들에 대해서 6가지 항생제에 대한 최소 억제 농도를 추가적으로 시험한 결과, 13 균주가 4가지 이상의 다른 계열의 항생제에 대한 다중 약제 내성인 것으로 나타났다. Isoelectric focusing gel electrophoresis에 의한 pI값과 DNA 염기서열을 분석한 결과 5균주가 TEM-1,1균주가 TEM-15,1 균주가 TEM-20,4균주가 TEM-52,2균주가 TEM-1과 AmpC, 1균주가 TEM-1과 OXA-30,1 균주가 TEM-1과 OXA-33,1 균주가 TEM-1, CTX-M-3, AmpC, 그리 고 1 균주가 AmpC를 생산하는 것으로 나타났으며, SHV를 생산하는 균주는 없었다. 이들의 항생제내성 유전자가 전달되는지 확인한 결과 동물로부터 분리된 대장균 (CCARM No.1203)에 extended-spectrum $\beta-lactamase$생산 유전자가 전달되는 것을 확인하였다. Random amplified polymorphic DNA와 pulsed field gel electrophoresis분석을 사용하여 genomic DNA에 대한 유전형을 분석한 결과 균주간의 유전적 연관성은 매우 낮은 것으로 나타나 한 병원에서 발견되는 균주는 clonal spread에 의한 것이라는 일반적인 보고와 다른 결과를 얻었다.

Keywords

References

  1. Bonnet, R., C. Chanal, E. Ageron, D. Sirot, C. De Champs, P. Grimont, and J. Sirot. 2002. Inducible AmpC $\beta$-lactamase of a new member Enterobacteriaceae. Antimicrob. Agents Chemother. 46, 3316-3319
  2. Bradford, P.A. 2001. Extended-spectrum $\beta$-lactamases in the 21st century. Clin. Microbiol. Rev. 114, 933-951
  3. Brinas, L., M. Zarazaga, Y. Senz, F. Ruiz-Larrea , and C. Torres. 2002. $\beta$-Lactamases in ampicillin-resistant Escherichia coli isolates from foods, humans, and healthy animals. Antimicrob. Agents Chemother. 246, 3156-3163
  4. Bush, K., G.A. Jacoby, and A.A. Medeiros. 1995. A functional classification scheme for $\beta$-lactamases and its correlation with molecular structures. Antimicrob. Agents Chemother. 39, 1211- 1233
  5. De Champs, C., D. Sirot, C. Chanal, R. Bonnet, J. Sirot, and the French Study Group. 2000. A 1998 Survey of extended-spectrum $\beta$-lactamases in Enterobacteriaceae in France. Antimicrob. Agents Chemother. 44, 3177-3179
  6. Emery, C.L. and L.A. Weymouth. 1997. Detection and clinical significance of extended-spectrum $\beta$-lactamases in a tertiary-care medical center. J. Clin. Microbiol. 35, 2061-2067
  7. Gniadkowski, M. and W. Hryniewicz. 1998. Outbreak of ceftazidime- resistant Klebsiella pneumoniae in a pediatric hospital in warsaw, Poland: Clonal spread of the TEM-47 extended spectrum $\beta$-lactamase (ESBL)-producing strain and transfer of a plasmid carrying the SHV-5-like ESBL- encoding gene. Antimicrob. Agents Chemother. 42, 3079-3085
  8. Gniadkowski, M., I. Schneider, A. Palucha, R. Jungwirth, B. Mikiewicz, and A. Bauernfeind. 1998. Cefotaxime-resistant Enterobacteriaceae isolates from a hospital in Warsaw, Poland: identification of a new CTX-M-3 cefotaxime-hydrolyzing beta-lactamase that is closely related to the CTX-M-1/MEN-1 enzyme. Antimicrob. Agents Chemother. 42, 827-832
  9. Jeong, S. H., I. K. Bae, S. G. Sohn, G. H. Kang, G. J. Jeon, Y. H. Kim, B. C. Jeong, and S. H. Lee. 2004. Molecular characterization of extended-spectrum beta-lactamases produced by clinical isolates of Klebsiella pneumoniae and Escherichia coli from a Korean nationwide survey. J Clin. Microbiol. 42, 2902-2906
  10. Kim, Y.K., H. Pai, H.J. Lee, S.E. Park, E.H. Choi, J. Kim, J.H. Kim, and E.C. Kim. 2002. Bloodstream infections by extended-spectrum $\beta$-lactamases-producing Escherichia coli and Klebsiella pneumoniae in children: epidemiology and clinical outcome. Antimicrob. Agents Chemother. 46, 1481-1491
  11. Lee, S.H., J.Y. Kim, S.H. Shin, Y.J. An, Y.W. Choi, Y.C. Jung, H.I. Jung, E.S. Sohn, S.H. Jeong, and K.J. Lee. 2003. Dissemination of SHV-12 and characterization of new AmpC-type $\beta$-lactamase genes among clinical isolates of Enterobacter species in Korea. J. Clin. Microbiol. 41, 2477-2482
  12. Livermore, D.M. 1995. $\beta$-Lctamases in laboratory and clinical resistance. Clin. Microbiol. Rev. 8, 557-584
  13. National Committee for Clinical Laboratory Standards. 2000. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically: Approved Standard-Fifth Edition. NCCLS. 20, 7-10
  14. Nelson, E.C., and B.G. Elisha. 1999. Molecular basis of AmpC hyperproduction in clinical isolates of Escherichia coli. Antimicrob. Agents Chemother. 43, 957-959
  15. Oliver, A., M. Prez-Vzquez, M. Martnez-Ferrer, F. Baquero, L. De Rafael, and R. Canton. 1999. Ampicillin-sulbactam and amoxicillin- clavulanate susceptibility testing of Escherichia coli isolates with different $\beta$-lactam resistance phenotypes. Antimicrob. Agents Chemother. 43, 862-867
  16. Pai, H., H.J. Lee, E.H. Choi, J. Kim, and G.A. Jacoby. 2001. Evolution of TEM-related extended-spectrum $\beta$-lactamases in Korea. Antimicrob. Agents Chemother. 45, 3651-3653
  17. Pai, H., S. Lyu, J.H. Lee, J. Kim, Y. Kwon, J.W. Kim, and K.W. Choe. 1999. Survey of extended-spectrum $\beta$-lactamases in clinical isolates of Escherichia coli and Klebsiella pneumoniae: prevalence of TEM-52 in Korea. J. Clin. Microbiol. 37, 1758-1763
  18. Perilli, M., E. Dell’Amico, B. Segatore, M.R. De Massis, C. Bianchi, F. Luzzaro, G.M. Rossolini, A. Toniolo, G. Nicoletti, and G. Amicosante. 2002. Molecular characterization of extended-spectrum $\beta$-lactamases produced by nosocomial isolates of Enterobacteriaceae from an Italian nationwide survey. J. Clin. Microbiol. 40, 611-614
  19. Philippon, A., G. Arlet, and G.A. Jacoby. 2002. Plasmid-determined AmpC-type $\beta$-lactamases. Antimicrob. Agents Chemother. 46, 1- 11
  20. Pitout, J.D., M.D. Reisbig, E.C. Venter, D.L. Church, and N.D. Hanson. 2003. Modification of the double-disk test for detection of Enterobacteriaceae producing extended-spectrum and AmpC $\beta$- lactamases. J. Clin. Microbiol. 41, 3933-3935
  21. Poirel, L., C. Heritier, V. Tolun, P. Nordmann. 2004. Emergence of oxacillinase-mediated resistance to imipenem in Klebsiella pneu moniae. Antimicrob. Agents Chemother. 48, 15-22
  22. Siu, L.K., P.L. Lu, J.Y. Chen, F.M. Lin, and S.C. Chang. 2003. High-level expression of AmpC $\beta$-lactamase due to insertion of nucleotides between -10 and -35 promoter sequences in Escherichia coli clinical isolates: Cases not responsive to extended-spectrum -cephalosporin treatment. Antimicrob. Agents Chemother. 47, 2138-2144
  23. Spanu, T., F. Luzzaro, M. Perilli, G. Amicosante, A. Toniolo, G. Fadda, and The Italian ESBL study group. 2002. Occurrence of extended-spectrum $\beta$-lctamases in members of the family Enterobacteriaceae in Italy: Implications for resistance to $\beta$-lactamase and other antimicrobial drugs. Antimicrob. Agents Chemother. 46, 196-202
  24. Sturenburg, E., D. Mack. 2003. Extended-spectrum $\beta$-lactamases: implications for the clinical microbiology laboratory, therapy, and infection control. J Infect. 47, 273-295
  25. Yan, J.J., W.C. Ko, S.H. Tsai, H.M. Wu, Y.T. Jin, and J.J. Wu. 2000. Dissemination of CTX-M-3 and CMY-2 $\beta$-lactamases among clinical isolates of Escherichia coli in Southern Taiwan. J. Clin. Microbiol. 38, 4320-4325
  26. Zhou, X.Y., F. Bordon, D. Sirot, M.D. Kitzis, and L. Gutmann. 1994. Emergence of clinical isolates of Escherichia coli producing TEM-1 derivatives or an OXA-1 beta-lactamase conferring resistance to beta-lactamase inhibitors. Antimicrob. Agents Chemother. 38, 1085-1089