Evaluation of EDTA-based Three Methods to Detect IMP-1 and VIM-2 Type Metallo-${\beta}$-Lactamase-Producing Clinical Isolates of Imipenem Resistant Acinetobacter and Pseudomonas spp.

  • Hong, Seung-Bok (Department of Clinical Laboratory Science, Juseong University) ;
  • Shin, Kyung-A (Department of Laboratory Medicine Bundang Jesaeng Hospital) ;
  • Hwang, Seock-Yeon (Department of Biomedical Laboratory Science, Daejeon University)
  • Received : 2011.02.28
  • Accepted : 2011.06.02
  • Published : 2011.06.30

Abstract

We compared three EDTA-based phenotypic screening methods for detecting IMP-1 and VIM-2 type metallo-${\beta}$- lactamase (MBL)-producing isolates of Acinetobacter and Pseudomonas spp., EDTA-double disk synergy test (EDTADDST), Etest MBL, and imipenem (IPM)-EDTA disk test. A total of 183 isolates (65 Acinetobacter spp. and 118 Pseudomonas spp. showing IPM resistance), confirmed to MBL genes by PCR, were used. The criteria for MBL production were (i) presence of a synergistic zone between IPM and EDTA disks in EDTA-DDST, (ii) reduction of IPM minimal inhibitory concentration by ${\geq}$ 3 twofold dilutions in the presence of EDTA in the Etest MBL, and (iii) ${\geq}$ 7 mm increase in the inhibition zone around the IPM plus EDTA disks compared with a sole IPM disk in the IPM-EDTA disk test. In this study using 87 MBL-producing and 96 MBL-nonproducing isolates, the sensitivities/specificities of EDTA-DDST, Etest MBL and IPM-EDTA disk tests were 94.3/78.1%, 89.7/91.7%, and 97.7/95.8%, respectively. When the threshold for the increase of the inhibition zone around the IPM plus EDTA disk over a sole IPM disk was altered to ${\geq}$ 5 mm and ${\geq}$ 8 mm for Acinetobacter spp. and Pseudomonas spp., respectively, the sensitivity and specificity of the test were 98.9% and 96.9%, respectively. Of the three EDTA-based phenotypic tests, the IMP-EDTA disk test was superior for detection of MBL-producing isolates.

Keywords

References

  1. Arakawa Y, Shibata N, Shibayama K, Kurokawa H, Yagi T, Fujiwara H, Goto M. Convenient test for screening metallo-$\beta$-lactamase-producing gram-negative bacteria by thiol compounds. J Clin Microbiol. 2000. 38: 40-43.
  2. Bush K, Jacoby JA, Medeiros AA. A functional classification scheme for $\beta$-lactamases and its correlation with molecular structure. Antimicrob Agents Chemother. 1995. 39: 1211-1233. https://doi.org/10.1128/AAC.39.6.1211
  3. Castanheira M, Toleman MA, Jones RN, Schmidt FJ, Walsh TR. Molecular characterization of a $\beta$-lactamase gene, $bla_{GIM-1}$, encoding a new subclass of metallo-$\beta$-lactamase. Antimicrob Agents Chemother. 2004. 48: 4654-4661. https://doi.org/10.1128/AAC.48.12.4654-4661.2004
  4. Chu YW, Cheung TKM, Ngan JYW, Kam KM. EDTA susceptibility leading to false detection of metallo-$\beta$-lactamase in Pseudomonas aeruginosa by Etest and an imipenem-EDTA disk method. Intern J Antimicrob Agents. 2005. 26: 338-341. https://doi.org/10.1016/j.ijantimicag.2005.07.001
  5. Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing. Sixteenth informational supplement. CLSI document M100-S9, Wayne, Pa. 2006.
  6. Hanson ND, Hossain A, Buck L, Moland ES, Thomson KS. First occurrence of a Pseudomonas aeruginosa isolate in the United States producing an IMP metallo-$\beta$-lactamase, IMP-18. Antimicrob Agents Chemother. 2006. 50: 2272-2273. https://doi.org/10.1128/AAC.01440-05
  7. Kim IS, Oh WI, Song JH, Lee NY. Screening and identification of metallo-$\beta$-lactamase gene in clinical isolates of imipenemresistant Pseudomonas aeruginosa. Korean J Lab Med. 2004. 24: 177-182.
  8. Kimura S, Ishii Y, Yamaguchi K. Evaluation of dipicolinic acid for detection of IMP- or VIM-type metallo-$\beta$-lactamase producing Pseudomonas aeruginosa clinical isolates. Diagn Microbiol Infect Dis. 2005. 53: 241-244. https://doi.org/10.1016/j.diagmicrobio.2005.05.017
  9. Lauretti L, Riccio ML, Mazzriol A, Cornaglia G, Amicosante G, Fontana R, Rossolini GM. Cloning and characterization of $bla_{VIM}$, a new integron-borne metallo-$\beta$-lactamase gene from a Pseudomonas aeruginosa clinical isolate. Antimicrob Agents Chemother. 1999. 43: 1584-1590.
  10. Lee K, Chong Y, Shin HB, Kim YA, Yong D, Yum JH. Modified Hodge and EDTA-disk synergy tests to screen metallo-$\beta$- lactamase-producing strains of Pseudomonas and Acinetobacter species. Clin Microbiol Infect. 2001. 7: 88-91. https://doi.org/10.1046/j.1469-0691.2001.00204.x
  11. Lee K, Lim YS, Yong D, Yum JH, Chong Y. Evaluation of the Hodge test and the imipenem-EDTA double-disk synergy test for differentiation metallo-$\beta$-lactamase-producing isolates of Pseudomonas spp. and Acinetobacter spp. J Clin Microbiol. 2003a. 41: 4623-4629. https://doi.org/10.1128/JCM.41.10.4623-4629.2003
  12. Lee K, Lee WG, Uh Y, Ha GY, Cho J, Chong Y. VIM-and IMP-type metallo-$\beta$-lactamase producing Pseudomonas spp. And Acinetobacter spp. in Korean Hospitals. Emerg Infect Dis. 2003b. 9: 868-871. https://doi.org/10.3201/eid09070.030012
  13. Lee K, Yum JH, Yong D, Lee HM, Kim HD, Docquier JD, Rossolini GM, Chong Y. Novel acquired metallo-$\beta$-lactamase gene, $bla_{SIM-1}$, in a class 1 integron from Acinetobacter baumannii clinical isolates from Korea. Antimicrob Agents Chemother. 2005a. 49: 4485-4491. https://doi.org/10.1128/AAC.49.11.4485-4491.2005
  14. Lee K, Yong D, Yum JH, Lim YS, Bolmstrom A, Qwarnstrom A, Karlsson A, Chong Y. Evaluation of Etest MBL for detection of $bla_{IMP-1}$ and $bla_{VIM-2}$ Allele-positive clinical isolates Pseudomonas spp. and Acinetobacter spp.. J Clin Microbiol. 2005b. 43: 942-944. https://doi.org/10.1128/JCM.43.2.942-944.2005
  15. Lee K, Kim MN, Choi TY, Cho SE, Lee S, Whang DH, Yong D, Chong Y, Woodford N, Livermore DM, the KONSAR Group. Wide dissemination of OXA-type carbapenemase in clinical Acinetobacter spp. isolates from South Korea. Int J Antimicrob Agents. 2009. 33: 520-524. https://doi.org/10.1016/j.ijantimicag.2008.10.009
  16. Marchiaro P, Mussi MA, Ballerini V, Pasteran F, Viale AM, Vila AJ, Limansky AS. Sensitive EDTA-based microbiological assays for detection of metallo-$\beta$-lactamases in nonfermentative gram-negative bacteria. J Clin Microbiol. 2005. 43: 5648-5652. https://doi.org/10.1128/JCM.43.11.5648-5652.2005
  17. Pitout JD, Chow BL, Gregson DB, Laupland KB, Elsayed S, Church DL. Molecular epidemiology of metallo-$\beta$-lactamaseproducing Pseudomonas aeruginosa in the Calgary Health Region; emergence of VIM-2 producing isolates. J Clin Microbiol. 2007. 45: 294-298. https://doi.org/10.1128/JCM.01694-06
  18. Poirel L, Naas T, Nicolas D, Collect L, Bellais S, Cavallo JD, Nordmann P. Characterization of VIM-2, a carbapenemhydrolyzing metallo-$\beta$-lactamase, and its plasmid- and integron-borne gene from a Pseudomonas aeruginosa clinical isolates in France. Antimicrob Agents Chemother. 2000. 44: 891-897. https://doi.org/10.1128/AAC.44.4.891-897.2000
  19. Riccio ML, Franceschini N, Boschi L, Caravelli B, Cornaglia G, Fontana R, Amicosante G, Rossolini GM. 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. 2000. 44: 1229-1235. https://doi.org/10.1128/AAC.44.5.1229-1235.2000
  20. Shin KS, Han K, Lee J, Hong SB, Son BR, Youn SJ, Kim J, Shin HS. Imipenem-resistant Achromobacter xylosoxidans carrying $bla_{VIM-2}$-containing class 1 integron. Diagn Microbiol Infect Dis. 2005. 53: 215-220. https://doi.org/10.1016/j.diagmicrobio.2005.06.018
  21. Toleman MA, Simm AM, Murphy TA, Gales AC, Biedenbach DJ, Jones RN, Walsh TR. Molecular characterization of SPM-1, a novel metallo-$\beta$-lactamase isolated in Latin America: report from the SENTRY antimicrobial programme. J Antimicrob Chemother. 2002. 50: 673-679. https://doi.org/10.1093/jac/dkf210
  22. Yong D, Lee K, Yum JH, Shin HB, Rossolini GM, Chong Y. Imipenem-EDTA disk method for differentiation of metallo-$\beta$-lactamase-producing clinical isolates of Pseudomonas spp. and Acinetobacter spp. J Clin Microbiol. 2002. 40: 3798-3801. https://doi.org/10.1128/JCM.40.10.3798-3801.2002
  23. Watanabe M, Iyobe S, Inoue M, Mitsuhashi S. Transferable imipenem resistance in Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1991. 35: 147-1451. https://doi.org/10.1128/AAC.35.1.147