title: International multicenter evaluation of the DiversiLab bacterial typing system for Escherichia coli and Klebsiella spp.
abstract: Successful multidrug-resistant clones are increasing in prevalence globally, which makes the ability to identify these clones urgent. However, adequate, easy-to-perform, and reproducible typing methods are lacking. We investigated whether DiversiLab (DL), an automated repetitive-sequence-based PCR bacterial typing system (bioMérieux), is suitable for comparing isolates analyzed at different geographic centers. A total of 39 Escherichia coli and 39 Klebsiella species isolates previously typed by the coordinating center were analyzed. Pulsed-field gel electrophoresis (PFGE) confirmed the presence of one cluster of 6 isolates, three clusters of 3 isolates, and three clusters of 2 isolates for each set of isolates. DL analysis was performed in 11 centers in six different countries using the same protocol. The DL profiles of 425 E. coli and 422 Klebsiella spp. were obtained. The DL system showed a lower discriminatory power for E. coli than did PFGE. The local DL data showed a low concordance, as indicated by the adjusted Rand and Wallace coefficients (0.132 to 0.740 and 0.070 to 1.0 [E. coli] and 0.091 to 0.864 and 0.056 to 1.0 [Klebsiella spp.], respectively). The central analysis showed a significantly improved concordance (0.473 to 1.0 and 0.290 to 1.0 [E. coli] and 0.513 to 0.965 and 0.425 to 1.0 [Klebsiella spp.], respectively). The misclassifications of profiles for individual isolates were mainly due to inconsistent amplification, which was most likely due to variations in the quality and amounts of the isolated DNA used for amplification. Despite local variations, the DL system has the potential to indicate the occurrence of clonal outbreaks in an international setting, provided there is strict adherence to standardized, reproducible DNA isolation methods and analysis protocols, all supported by a central database for profile comparisons.
authors.affiliation: Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, the Netherlands.
authors.forename: Guido M
authors.initials: GM
authors.lastname: Voets
authors.forename: Maurine A
authors.initials: MA
authors.lastname: Leverstein-van Hall
authors.forename: Susanne
authors.initials: S
authors.lastname: Kolbe-Busch
authors.forename: Adri
authors.initials: A
authors.lastname: van der Zanden
authors.forename: Deirdre
authors.initials: D
authors.lastname: Church
authors.forename: Martin
authors.initials: M
authors.lastname: Kaase
authors.forename: Andrea
authors.initials: A
authors.lastname: Grisold
authors.forename: Mathew
authors.initials: M
authors.lastname: Upton
authors.forename: Elaine
authors.initials: E
authors.lastname: Cloutman-Green
authors.forename: Rafael
authors.initials: R
authors.lastname: Cantón
authors.forename: Alexander W
authors.initials: AW
authors.lastname: Friedrich
authors.forename: Ad C
authors.initials: AC
authors.lastname: Fluit
mesh_terms: D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000145:classification* / Q000235:genetics; D006801:Humans; D007391:International Cooperation; D007709:Klebsiella / Q000145:classification* / Q000235:genetics; D058889:Molecular Typing / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results
chemical_list: D004269:DNA, Bacterial
references.citation: Clin Microbiol Infect. 2014 Jan;20(1):34-7
references.pmid: 23464880
references.citation: J Clin Microbiol. 2001 Nov;39(11):4190-2
references.pmid: 11682558
references.citation: Antimicrob Agents Chemother. 2010 Sep;54(9):3564-8
references.pmid: 20547788
references.citation: J Clin Microbiol. 1994 Feb;32(2):407-15
references.pmid: 7908673
references.citation: Nucleic Acids Res. 1991 Dec 25;19(24):6823-31
references.pmid: 1762913
references.citation: J Clin Microbiol. 2010 Nov;48(11):3979-89
references.pmid: 20861340
references.citation: Euro Surveill. 2013 Jan 24;18(4):20380
references.pmid: 23369389
references.citation: Clin Microbiol Infect. 2010 Mar;16(3):232-7
references.pmid: 19416293
references.citation: J Antimicrob Chemother. 2009 Aug;64(2):348-52
references.pmid: 19525514
references.citation: J Microbiol Methods. 2010 Nov;83(2):224-30
references.pmid: 20849889
references.citation: PLoS One. 2012;7(3):e34294
references.pmid: 22457832
references.citation: Antimicrob Agents Chemother. 2009 Feb;53(2):818-20
references.pmid: 19029323
references.citation: PLoS One. 2012;7(12):e52102
references.pmid: 23284886
references.citation: J Clin Microbiol. 2009 Apr;47(4):1212-5
references.pmid: 19204095
references.citation: Trends Microbiol. 2002;10(10 Suppl):S15-21
references.pmid: 12377563
references.citation: J Antimicrob Chemother. 2011 Jul;66(7):1510-3
references.pmid: 21543359
references.citation: J Clin Microbiol. 2009 Aug;47(8):2452-7
references.pmid: 19553588
references.citation: Clin Infect Dis. 2011 Jul 1;53(1):49-56
references.pmid: 21653302
references.citation: Clin Microbiol Infect. 2007 Oct;13 Suppl 3:1-46
references.pmid: 17716294
references.citation: J Microbiol Methods. 2011 Jan;84(1):74-80
references.pmid: 21062634
references.citation: J Clin Microbiol. 1988 Nov;26(11):2465-6
references.pmid: 3069867
references.citation: J Microbiol Methods. 2009 Apr;77(1):130-3
references.pmid: 19318062
references.citation: J Clin Microbiol. 2005 Mar;43(3):1187-92
references.pmid: 15750081