Multi-centre evaluation of real-time multiplex PCR for detection of carbapenemase genes OXA-48, VIM, IMP, NDM and KPC
Tóm tắt
Resistance to carbapenem antibiotics is emerging worldwide among Enterobacteriaceae. To prevent hospital transmission due to unnoticed carriage of carbapenemase producing micro-organisms in newly admitted patients, or follow-up of patients in an outbreak setting, a molecular screening method was developed for detection of the most prevalent carbapenemase genes; blaOXA-48, blaVIM, blaIMP, blaNDM and blaKPC. A real-time multiplex PCR assay was evaluated using a collection of 86 Gram negative isolates, including 62 carbapenemase producers. Seven different laboratories carried out this method and used the assay for detection of the carbapenemase genes on a selection of 20 isolates. Both sensitivity and specificity of the multiplex PCR assay was 100%, as established by results on the strain collection and the inter-laboratory comparisons. In this study, we present a multiplex real-time PCR that is a robust, reliable and rapid method for the detection of the most prevalent carbapenemases blaOXA-48, blaVIM, blaIMP, blaNDM and blaKPC, and is suitable for screening of broth cultured rectal swabs and for identification of carbapenemase genes in cultures.
Tài liệu tham khảo
Cuzon G, Ouanich J, Gondret R, Naas T, Nordmann P: Outbreak of OXA-48-positive carbapenem-resistant Klebsiella pneumoniae isolates in France. Antimicrob Agents Chemother. 2011, 55: 2420-2423. 10.1128/AAC.01452-10.
Goren MG, Chmelnitsky I, Carmeli Y, Navon-Venezia S: Plasmid-encoded OXA-48 carbapenemase in Escherichia coli from Israel. J Antimicrob Chemother. 2011, 66 (3): 672-673. 10.1093/jac/dkq467.
Kitchel B, Rasheed JK, Patel JB, Srinivasan A, Navon-Venezia S, Carmeli Y, Brolund A, Giske CG: Molecular Epidemiology of KPC-Producing Klebsiella pneumoniae Isolates in the United States: Clonal Expansion of Multilocus Sequence Type 258. Antimicrob Agents Chemother. 2009, 53 (8): 3365-3370. 10.1128/AAC.00126-09.
Girlich D, Poirel L, Nordmann P: Value of the modified hodge test for detection of emerging carbapenemases in Enterobacteriaceae. J Clin Microbiol. 2012, 50: 477-479. 10.1128/JCM.05247-11.
Giske CG, Gezelius L, Samuelsen Ø, Warner M, Sundsfjord A, Woodford N: A sensitive and specific phenotypic assay for detection of metallo-β-lactamases and KPC in Klebsiella pneumoniae with the use of meropenem disks supplemented with aminophenylboronic acid, dipicolinic acid and cloxacillin. Clin Microbiol Infect. 2011, 17 (4): 552-556. 10.1111/j.1469-0691.2010.03294.x.
Pasteran F, Mendez T, Guerriero L, Rapoport M, Corso A: Sensitive screening tests for suspected class A carbapenemase production in species of Enterobacteriaceae. J Clin Microbiol. 2009, 47: 1631-1639. 10.1128/JCM.00130-09.
Tsakris A, Poulou A, Pournaras S, Voulgari E, Vrioni G, Themeli-Digalaki K, Petropoulou D, Sofianou D: A simple phenotypic method for the differentiation of metallo-beta-lactamases and class A KPC carbapenemases in Enterobacteriaceae clinical isolates. J Antimicrob Chemother. 2010, 65 (8): 664-671.
Cohen Stuart J, Leverstein-Van Hall MA, on behalf of members of the Dutch Working Party on the Detection of Highly Resistant Microorganisms: Guideline for phenotypic screening and confirmation of carbapenemases in Enterobacteriaceae. Int J Antimicrob Agents. 2010, 36: 205-210. 10.1016/j.ijantimicag.2010.05.014.
Van Hannen EJ: Nieuwegein, The Netherlands: St. Antonius Hospital, [email protected]. Personal communication
Voets GM, Fluit AC, Scharringa J, Cohen Stuart J, Leverstein-van Hall MA: A set of multiplex PCRs for genotypic detection of extended-spectrum beta-lactamases, carbapenemases, plasmid-mediated AmpC 1-lactamases and OXA-lactamases. Int J Antimicrobial Agents. 2011, 37: 356-359. 10.1016/j.ijantimicag.2011.01.005.
Chen L, Mediavilla JR, Endimiani A, Rosenthal ME, Zhao Y, Bonomo RA, Kreiswirth BN: Multiplex Real-Time PCR Assay for Detection and Classification of Klebsiella pneumoniae Carbapenemase Gene (blaKPC) Variants. J Clin Microbiol. 2011, 49 (2): 579-585. 10.1128/JCM.01588-10.
Cole JM, Schuetz AN, Hill CE, Nolte FS: Development and Evaluation of a Real-Time PCR Assay for Detection of Klebsiella pneumoniae Carbapenemase Genes. J Clin Microbiol. 2009, 47 (2): 322-326. 10.1128/JCM.01550-08.
Naas T, Ergani A, Carrër A, Nordmann P: Real-Time PCR for Detection of NDM-1 Carbapenemase Genes from Spiked Stool Samples. Antimicrob Agents Chemother. 2011, 55 (9): 4038-4043. 10.1128/AAC.01734-10.
Poirel L, Walsh TR, Cuvillier V, Nordmann P: Multiplex PCR for detection of acquired carbapenemase genes. Diagn Microbiol Infect Dis. 2011, 70: 119-123. 10.1016/j.diagmicrobio.2010.12.002.
Van der Bij AK, Mol M, Van Westreenen M, Goessens WHF, Pitout JDD: The laboratory diagnosis of Pseudomonas aeruginosa that produce metallo- beta -lactamases in a Dutch tertiary care centre. Scand J Infect Dis. 2011, 43 (8): 596-602. 10.3109/00365548.2011.574148.
The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2334/14/27/prepub