ORCID
- Upton, Mathew: 0000-0003-4287-6396
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.
DOI
10.1128/JCM.01664-13
Publication Date
2013-12-01
Publication Title
J Clin Microbiol
Volume
51
Issue
12
Organisational Unit
School of Biomedical Sciences
Keywords
DNA, Bacterial, Escherichia coli, Humans, International Cooperation, Klebsiella, Molecular Typing, Polymerase Chain Reaction, Reproducibility of Results
First Page
3944
Last Page
3949
Recommended Citation
Voets, G. M., Leverstein-van, H., Kolbe-Busch, S., van, d., Church, D., Kaase, M., Grisold, A., Upton, M., Cloutman-Green, E., Cantón, R., Friedrich, A., Fluit, A., & Group, D. (2013) 'International multicenter evaluation of the DiversiLab bacterial typing system for Escherichia coli and Klebsiella spp.', J Clin Microbiol, 51(12), pp. 3944-3949. Available at: https://doi.org/10.1128/JCM.01664-13
