Metallo-beta-lactamase Detection: Comparative Evaluation of Double-disk Synergy Versus Combined Disk Tests for IMP-, GIM-, SIM-, SPM-, or VIM-producing Isolates

Overview

Journal J Clin Microbiol

Specialty Microbiology

Date 2008 Mar 7

PMID 18322055

Citations 38

Authors

Renata C Picao

Soraya S Andrade

Adriana Gianinni Nicoletti

Eloiza H Campana

Gabriela C Moraes

Rodrigo E Mendes

Ana C Gales

Affiliations

Soon will be listed here.

Abstract

The emergence of metallo-beta-lactamase (MBL)-producing isolates is a challenge to routine microbiology laboratories, since there are no standardized methods for detecting such isolates. The aim of this study was to evaluate the accuracy of different phenotypic methods to detect MBL production among Pseudomonas spp., Acinetobacter spp., and enterobacterial isolates, including GIM, IMP, SIM, SPM, and VIM variants. A total of 46 genetically unrelated Pseudomonas aeruginosa, Pseudomonas putida, Acinetobacter sp., and enterobacterial strains producing distinct MBLs were tested. Nineteen strains were included as negative controls. The inhibition of bacterial growth and beta-lactam hydrolysis caused by MBL inhibitors (IMBL) also were evaluated. The isolates were tested for MBL production by both a double-disk synergy test (DDST) and a combined disk assay (CD) using imipenem and ceftazidime as substrates in combination with distinct IMBL. One hundred percent sensitivity and specificity were achieved by DDST using 2-mercaptopropionic acid in combination with ceftazidime and imipenem for the detection of MBL production among P. aeruginosa and Acinetobacter species isolates, respectively. The CD test showed the same results for detecting MBL-producing enterobacteria by combining imipenem and EDTA, with a 5.0-mm-breakpoint increase in the size of the inhibition zone. Our results indicate that both phenotypic methods to detect MBL-producing isolates should be based on the genera to be tested, regardless of the enzyme produced by such isolates, as well as on the local prevalence of MBL producers.

Citing Articles

Phenotypic identification of Metallo-ß- lactamase resistance Gram negative bacteria from a clinical specimen in Sidama, Ethiopia.

Alemayehu T, Abera W, Ali M, Jimma B, Ayalew H, Habte L PLoS One. 2025; 20(1):e0313431.

PMID: 39775289 PMC: 11709301. DOI: 10.1371/journal.pone.0313431.

Genotypic characterization and clonal relatedness of metallo-β-lactamase-producing non-fermentative gram negative bacteria in the first 5 years of their circulation in Paraguay (2011-2015).

Franco R, de Oliveira Santos I, Mora M, Lopez P, Alvarez V, Arce F Braz J Microbiol. 2022; 54(1):179-190.

PMID: 36564646 PMC: 9943808. DOI: 10.1007/s42770-022-00888-x.

Novel Strains Co-Harbouring Metallo β-Lactamase and Isolated from Immunocompromised Paediatric Patients.

Chen H, Mai H, Lopes B, Wen F, Patil S Infect Drug Resist. 2022; 15:2929-2936.

PMID: 35706928 PMC: 9189156. DOI: 10.2147/IDR.S368566.

Detection of metallo-beta lactamases and oxacillinase genes in carbapenem-resistant strains isolated in Morocco.

Massik A, Hibaoui L, Arhoune B, Yahyaoui G, Oumokhtar B, Mahmoud M Pan Afr Med J. 2022; 40:210.

PMID: 35136473 PMC: 8783299. DOI: 10.11604/pamj.2021.40.210.28663.

Virulence characterization and clonal analysis of uropathogenic Escherichia coli metallo-beta-lactamase-producing isolates.

Zangane Matin F, Rezatofighi S, Roayaei Ardakani M, Akhoond M, Mahmoodi F Ann Clin Microbiol Antimicrob. 2021; 20(1):50.

PMID: 34344363 PMC: 8336094. DOI: 10.1186/s12941-021-00457-4.

References

Marchiaro P, Mussi M, Ballerini V, Pasteran F, Viale A, Vila A . Sensitive EDTA-based microbiological assays for detection of metallo-{beta}-lactamases in nonfermentative gram-negative bacteria. J Clin Microbiol. 2005; 43(11):5648-52. PMC: 1287843. DOI: 10.1128/JCM.43.11.5648-5652.2005. View

Sader H, Castanheira M, Mendes R, Toleman M, Walsh T, Jones R . Dissemination and diversity of metallo-beta-lactamases in Latin America: report from the SENTRY Antimicrobial Surveillance Program. Int J Antimicrob Agents. 2004; 25(1):57-61. DOI: 10.1016/j.ijantimicag.2004.08.013. View

Petropoulou D, Tzanetou K, Syriopoulou V, Daikos G, Ganteris G, Malamou-Lada E . Evaluation of imipenem/imipenem+EDTA disk method for detection of metallo-beta-lactamase-producing Klebsiella pneumoniae isolated from blood cultures. Microb Drug Resist. 2006; 12(1):39-43. DOI: 10.1089/mdr.2006.12.39. View

Gales A, Biedenbach D, Winokur P, Hacek D, Pfaller M, Jones R . Carbapenem-resistant Serratia marcescens isolates producing Bush group 2f beta-lactamase (SME-1) in the United States: results from the MYSTIC Programme. Diagn Microbiol Infect Dis. 2001; 39(2):125-7. DOI: 10.1016/s0732-8893(00)00222-4. View

Chu Y, Cheung T, Ngan J, Kam K . EDTA susceptibility leading to false detection of metallo-beta-lactamase in Pseudomonas aeruginosa by Etest and an imipenem-EDTA disk method. Int J Antimicrob Agents. 2005; 26(4):340-1. DOI: 10.1016/j.ijantimicag.2005.07.004. View

Tognim M, Gales A, Penteado A, Silbert S, Sader H . Dissemination of IMP-1 metallo- beta -lactamase-producing Acinetobacter species in a Brazilian teaching hospital. Infect Control Hosp Epidemiol. 2006; 27(7):742-7. DOI: 10.1086/504356. View

Jones R, Deshpande L, Bell J, Turnidge J, Kohno S, Hirakata Y . Evaluation of the contemporary occurrence rates of metallo-beta-lactamases in multidrug-resistant Gram-negative bacilli in Japan: report from the SENTRY Antimicrobial Surveillance Program (1998-2002). Diagn Microbiol Infect Dis. 2004; 49(4):289-94. DOI: 10.1016/j.diagmicrobio.2004.04.007. View

Zavascki A, Barth A, Goncalves A, Moro A, Fernandes J, Martins A . The influence of metallo-beta-lactamase production on mortality in nosocomial Pseudomonas aeruginosa infections. J Antimicrob Chemother. 2006; 58(2):387-92. DOI: 10.1093/jac/dkl239. View

Migliavacca R, Docquier J, Mugnaioli C, Amicosante G, Daturi R, Lee K . Simple microdilution test for detection of metallo-beta-lactamase production in Pseudomonas aeruginosa. J Clin Microbiol. 2002; 40(11):4388-90. PMC: 139638. DOI: 10.1128/JCM.40.11.4388-4390.2002. View

10.

Toleman M, Rolston K, Jones R, Walsh T . blaVIM-7, an evolutionarily distinct metallo-beta-lactamase gene in a Pseudomonas aeruginosa isolate from the United States. Antimicrob Agents Chemother. 2003; 48(1):329-32. PMC: 310168. DOI: 10.1128/AAC.48.1.329-332.2004. View

11.

Siemann S, Clarke A, Viswanatha T, Dmitrienko G . Thiols as classical and slow-binding inhibitors of IMP-1 and other binuclear metallo-beta-lactamases. Biochemistry. 2003; 42(6):1673-83. DOI: 10.1021/bi027072i. View

12.

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(3):241-4. DOI: 10.1016/j.diagmicrobio.2005.05.017. View

13.

Yong D, Lee K, Yum J, Shin H, Rossolini G, 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(10):3798-801. PMC: 130862. DOI: 10.1128/JCM.40.10.3798-3801.2002. View

14.

Zavascki A, Gaspareto P, Martins A, Goncalves A, Barth A . Outbreak of carbapenem-resistant Pseudomonas aeruginosa producing SPM-1 metallo-{beta}-lactamase in a teaching hospital in southern Brazil. J Antimicrob Chemother. 2005; 56(6):1148-51. DOI: 10.1093/jac/dki390. View

15.

Lincopan N, Mcculloch J, Reinert C, Cassettari V, Gales A, Mamizuka E . First isolation of metallo-beta-lactamase-producing multiresistant Klebsiella pneumoniae from a patient in Brazil. J Clin Microbiol. 2005; 43(1):516-9. PMC: 540118. DOI: 10.1128/JCM.43.1.516-519.2005. View

16.

Castanheira M, Toleman M, Jones R, Schmidt F, Walsh T . Molecular characterization of a beta-lactamase gene, blaGIM-1, encoding a new subclass of metallo-beta-lactamase. Antimicrob Agents Chemother. 2004; 48(12):4654-61. PMC: 529189. DOI: 10.1128/AAC.48.12.4654-4661.2004. View

17.

Linden A . Measuring diagnostic and predictive accuracy in disease management: an introduction to receiver operating characteristic (ROC) analysis. J Eval Clin Pract. 2006; 12(2):132-9. DOI: 10.1111/j.1365-2753.2005.00598.x. View

18.

Arakawa Y, Shibata N, Shibayama K, Kurokawa H, Yagi T, Fujiwara H . Convenient test for screening metallo-beta-lactamase-producing gram-negative bacteria by using thiol compounds. J Clin Microbiol. 2000; 38(1):40-3. PMC: 86013. DOI: 10.1128/JCM.38.1.40-43.2000. View

19.

Lee K, Yong D, Yum J, Lim Y, Bolmstrom A, Qwarnstrom A . Evaluation of Etest MBL for detection of blaIMP-1 and blaVIM-2 allele-positive clinical isolates of Pseudomonas spp. and Acinetobacter spp. J Clin Microbiol. 2005; 43(2):942-4. PMC: 548058. DOI: 10.1128/JCM.43.2.942-944.2005. View

20.

Toleman M, Simm A, Murphy T, Gales A, Biedenbach D, Jones R . Molecular characterization of SPM-1, a novel metallo-beta-lactamase isolated in Latin America: report from the SENTRY antimicrobial surveillance programme. J Antimicrob Chemother. 2002; 50(5):673-9. DOI: 10.1093/jac/dkf210. View