Abstract
The Mast® D68C test is a phenotypical test that allows the detection of extended-spectrum β-lactamase (ESBL) production, even in AmpC-producing Enterobacteriaceae. We assessed its detection accuracy against a large collection of 106 Enterobacteriaceae isolates producing a wide diversity of well-characterized β-lactamases (53 ESBL producers, 25 Amp. producers, seven AmpC and ESBL producers, five carbapenemase producers, three carbapenemase and ESBL producers, one AmpC, carbapenemase, and ESBL producer, three TEM-1 producers, three SHV-1 producers, three OXA-1 producers, and one hyperOXY producer, ATCC 35218, ATCC 25922 [a β-lactamase-negative control strain]). The results were compared with those of the double disk test and the Clinical and Laboratory Standards Institute (CLSI) confirmatory test for the detection of ESBL. The sensitivity was 90.6 % for the synergy test, 87.5 % for the CLSI method, and only 73.1 % for D68C, which, however, reached 92.1 % if the strains for which supplementary investigations were recommended and the complex mutant TEM (CMT)-producing strains were excluded versus 94.1 % and 88.2 % for the other methods. The specificity was 90.2 % for the synergy test and 100 % for the CLSI method and D68C. D68C was also efficient in detecting AmpC-overproducing strains (sensitivity = 97 %, specificity = 95.9 %): among the 74 strains belonging to natural AmpC-producing species, the sensitivity and specificity were 100 and 94.8 %, respectively. The Mast® D68C-test is a promising method that is easy to perform for the detection of current ESBLs and could also be useful for the detection of plasmid-encoded AmpC enzymes (sensitivity = 100 %).
Similar content being viewed by others
References
Bradford PA (2001) Extended-spectrum beta-lactamases in the 21st century: characterization, epidemiology, and detection of this important resistance threat. Clin Microbiol Rev 14(4):933–951, table of contents
Sirot D, Sirot J, Labia R, Morand A, Courvalin P, Darfeuille-Michaud A, Perroux R, Cluzel R (1987) Transferable resistance to third-generation cephalosporins in clinical isolates of Klebsiella pneumoniae: identification of CTX-1, a novel beta-lactamase. J Antimicrob Chemother 20(3):323–334
Paterson DL, Bonomo RA (2005) Extended-spectrum beta-lactamases: a clinical update. Clin Microbiol Rev 18(4):657–686
Bonnet R (2004) Growing group of extended-spectrum beta-lactamases: the CTX-M enzymes. Antimicrob Agents Chemother 48(1):1–14
Naas T, Poirel L, Karim A, Nordmann P (1999) Molecular characterization of In50, a class 1 integron encoding the gene for the extended-spectrum beta-lactamase VEB-1 in Pseudomonas aeruginosa. FEMS Microbiol Lett 176(2):411–419
Nordmann P, Naas T (1994) Sequence analysis of PER-1 extended-spectrum beta-lactamase from Pseudomonas aeruginosa and comparison with class A beta-lactamases. Antimicrob Agents Chemother 38(1):104–114
Poirel L, Le Thomas I, Naas T, Karim A, Nordmann P (2000) Biochemical sequence analyses of GES-1, a novel class A extended-spectrum beta-lactamase, and the class 1 integron In52 from Klebsiella pneumoniae. Antimicrob Agents Chemother 44(3):622–632
Robin F, Delmas J, Schweitzer C, Bonnet R (2008) Evaluation of the Vitek-2 extended-spectrum beta-lactamase test against non-duplicate strains of Enterobacteriaceae producing a broad diversity of well-characterised beta-lactamases. Clin Microbiol Infect 14(2):148–154
Howard SJ, Lass-Flörl C, Cuenca-Estrella M, Gomez-Lopez A, Arendrup MC (2013) Determination of isavuconazole susceptibility of Aspergillus and Candida species by the EUCAST method. Antimicrob Agents Chemother 57(11):5426–5431
European Committee on Antimicrobial Susceptibility Testing (EUCAST) (2013) EUCAST guideline for the detection of resistance mechanisms and specific resistances of clinical and/or epidemiological importance. Available online at: http://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Resistance_mechanisms/EUCAST_detection_of_resistance_mechanisms_v1.0_20131211.pdf
Pérez-Pérez FJ, Hanson ND (2002) Detection of plasmid-mediated AmpC beta-lactamase genes in clinical isolates by using multiplex PCR. J Clin Microbiol 40(6):2153–2162
Miró E, Agüero J, Larrosa MN, Fernández A, Conejo MC, Bou G, González-López JJ, Lara N, Martínez-Martínez L, Oliver A, Aracil B, Oteo J, Pascual A, Rodríguez-Baño J, Zamorano L, Navarro F (2013) Prevalence and molecular epidemiology of acquired AmpC beta-lactamases and carbapenemases in Enterobacteriaceae isolates from 35 hospitals in Spain. Eur J Clin Microbiol Infect Dis 32(2):253–259
Comité de l’Antibiogramme de la Société Française de Microbiologie (2007) Communiqué 2007
Clinical and Laboratory Standards Institute (CLSI) (2014) Performance standards for antimicrobial susceptibility testing; Twenty-fourth informational supplement update. CLSI document M100-S24. CLSI, Wayne, PA
Beceiro A, Maharjan S, Gaulton T, Doumith M, Soares NC, Dhanji H, Warner M, Doyle M, Hickey M, Downie G, Bou G, Livermore DM, Woodford N (2011) False extended-spectrum {beta}-lactamase phenotype in clinical isolates of Escherichia coli associated with increased expression of OXA-1 or TEM-1 penicillinases and loss of porins. J Antimicrob Chemother 66(9):2006–2010
Poirel L, Mammeri H, Nordmann P (2004) TEM-121, a novel complex mutant of TEM-type beta-lactamase from Enterobacter aerogenes. Antimicrob Agents Chemother 48(12):4528–4531
Robin F, Delmas J, Archambaud M, Schweitzer C, Chanal C, Bonnet R (2006) CMT-type beta-lactamase TEM-125, an emerging problem for extended-spectrum beta-lactamase detection. Antimicrob Agents Chemother 50(7):2403–2408
Robin F, Delmas J, Schweitzer C, Tournilhac O, Lesens O, Chanal C, Bonnet R (2007) Evolution of TEM-type enzymes: biochemical and genetic characterization of two new complex mutant TEM enzymes, TEM-151 and TEM-152, from a single patient. Antimicrob Agents Chemother 51(4):1304–1309
Ingram PR, Inglis TJ, Vanzetti TR, Henderson BA, Harnett GB, Murray RJ (2011) Comparison of methods for AmpC beta-lactamase detection in Enterobacteriaceae. J Med Microbiol 60(Pt 6):715–721
Mavroidi A, Tzelepi E, Tsakris A, Miriagou V, Sofianou D, Tzouvelekis LS (2001) An integron-associated beta-lactamase (IBC-2) from Pseudomonas aeruginosa is a variant of the extended-spectrum beta-lactamase IBC-1. J Antimicrob Chemother 48(5):627–630
Giakkoupi P, Tzouvelekis LS, Tsakris A, Loukova V, Sofianou D, Tzelepi E (2000) IBC-1, a novel integron-associated class A beta-lactamase with extended-spectrum properties produced by an Enterobacter cloacae clinical strain. Antimicrob Agents Chemother 44(9):2247–2253
Polsfuss S, Bloemberg GV, Giger J, Meyer V, Böttger EC, Hombach M (2012) Evaluation of a diagnostic flow chart for detection and confirmation of extended spectrum beta-lactamases (ESBL) in Enterobacteriaceae. Clin Microbiol Infect 18(12):1194–1204
Oliver A, Weigel LM, Rasheed JK, McGowan JE Jr, Raney P, Tenover FC (2002) Mechanisms of decreased susceptibility to cefpodoxime in Escherichia coli. Antimicrob Agents Chemother 46(12):3829–3836
Jacoby GA (2009) AmpC beta-lactamases. Clin Microbiol Rev 22(1):161–182, Table of Contents
Peter-Getzlaff S, Polsfuss S, Poledica M, Hombach M, Giger J, Böttger EC, Zbinden R, Bloemberg GV (2011) Detection of AmpC beta-lactamase in Escherichia coli: comparison of three phenotypic confirmation assays and genetic analysis. J Clin Microbiol 49(8):2924–2932
Polsfuss S, Bloemberg GV, Giger J, Meyer V, Böttger EC, Hombach M (2011) Practical approach for reliable detection of AmpC beta-lactamase-producing Enterobacteriaceae. J Clin Microbiol 49(8):2798–2803
Sabia C, Gargiulo R, Sarti M (2012) Evaluation of a double synergy differential test (DSDT) for differential detection of ESBL and AmpC-type beta-lactamases in Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis. New Microbiol 35(2):221–225
Acknowledgments
We thank Laurent Guillouard, Marlene Jan, and Rolande Perroux for the technical assistance. We thank Mast® Diagnostic for providing the D68C tests. This work was supported in part by a grant from the Ministère de l’Éducation Nationale, Ministère de l’Enseignement Supérieur et de la Recherche, Paris, France.
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval
Not required.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Nourrisson, C., Tan, R.N., Hennequin, C. et al. The MAST® D68C test: an interesting tool for detecting extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae. Eur J Clin Microbiol Infect Dis 34, 975–983 (2015). https://doi.org/10.1007/s10096-014-2305-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10096-014-2305-6