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J Clin Microbiol. 2015 Feb;53(2):498-503. doi: 10.1128/JCM.01611-14. Epub 2014 Nov 26.

High-throughput mycobacterial interspersed repetitive-unit-variable-number tandem-repeat genotyping for Mycobacterium tuberculosis epidemiological studies.

Author information

1
Laboratoire des Pathogènes Emergents (LPE), Fondation Mérieux-Centre International de Recherche en Infectiologie (CIRI), Lyon, France marie.gauthier@fondation-merieux.org.
2
Laboratoire des Pathogènes Emergents (LPE), Fondation Mérieux-Centre International de Recherche en Infectiologie (CIRI), Lyon, France.
3
National Center for Tuberculosis and Lung Disease (NCTLD), Tbilisi, Georgia.
4
Centre d'Infectiologie Christophe Mérieux du Laos (CICML), Vientiane, Lao People's Democratic Republic.
5
Les Centres GHESKIO, Port-au-Prince, Haiti.

Abstract

The emergence of drug-resistant forms of tuberculosis (TB) represents a major public health concern. Understanding the transmission routes of the disease is a key factor for its control and for the implementation of efficient interventions. Mycobacterial interspersed repetitive-unit-variable-number tandem-repeat (MIRU-VNTR) marker typing is a well-described method for lineage identification and transmission tracking. However, the conventional manual genotyping technique is cumbersome and time-consuming and entails many risks for errors, thus hindering its implementation and dissemination. We describe here a new approach using the QIAxcel system, an automated high-throughput capillary electrophoresis system that also carries out allele calling. This automated method was assessed on 1,824 amplicons from 82 TB isolates and tested with sets of markers of 15 or 24 loci. Overall allele-calling concordance between the methods from 140 to 1,317 bp was 98.9%. DNA concentrations and repeatability and reproducibility performances showed no biases in allele calling. Furthermore, turnaround time using this automated system was reduced by 81% compared to the conventional manual agarose gel method. In sum, this new automated method facilitates MIRU-VNTR genotyping and provides reliable results. Therefore, it is well suited for field genotyping. The implementation of this method will help to achieve accurate and cost-effective epidemiological studies, especially in countries with a high prevalence of TB, where the high number of strains complicates the surveillance of circulating lineages and requires efficient interventions to be carried out in an urgent manner.

PMID:
25428144
PMCID:
PMC4298502
DOI:
10.1128/JCM.01611-14
[Indexed for MEDLINE]
Free PMC Article

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