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Journal of Clinical Microbiology, December 2008, p. 3935-3940, Vol. 46, No. 12
0095-1137/08/$08.00+0     doi:10.1128/JCM.00464-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Evaluation of a Multilocus Variable-Number Tandem-Repeat Analysis Scheme for Typing Human Brucella Isolates in a Region of Brucellosis Endemicity{triangledown} ,{dagger}

Mireille M. Kattar,1,2* Rola F. Jaafar,3 George F. Araj,1,2 Philippe Le Flèche,4,5 Ghassan M. Matar,3 Roland Abi Rached,1,2 Simon Khalife,1,2 and Gilles Vergnaud4,6*

Department of Pathology and Laboratory Medicine,1 Department of Microbiology and Immunology, American University of Beirut, Beirut, Lebanon,3 World Health Organization Regional Collaborating Center on Human Brucellosis, Beirut, Lebanon,2 Université Paris-Sud 11, CNRS, UMR8621, Institut de Génétique et Microbiologie, Orsay 91405, France,4 Department of Analytical Microbiology, Centre d'Etudes du Bouchet, Vert le Petit 91710, France,5 DGA/D4S-Mission pour la Recherche et l'Innovation Scientifique, Bagneux 92220, France6

Received 9 March 2008/ Returned for modification 6 July 2008/ Accepted 30 September 2008

Brucellosis remains an important anthropozoonosis worldwide. Brucella species are genetically homogeneous, and thus, the typing of Brucella species for epidemiological purposes by conventional molecular typing methods has remained elusive. Although many methods could segregate isolates into the phylogenetically recognized taxa, limited within-species genetic diversity has been identified. Recently, multilocus variable-number tandem-repeat analysis (MLVA) was found to have a high degree of resolution when it was applied to collections of Brucella isolates from geographically widespread locations, and an assay comprising 16 such loci (MLVA-16) was proposed. This scheme includes eight minisatellite loci (panel 1) and eight microsatellites (panel 2, which is subdivided into panels 2A and 2B). The utility of MLVA-16 for the subtyping of human Brucella isolates from geographically restricted regions needs to be further evaluated, and genotyping databases with worldwide coverage must be progressively established. In the present study, MLVA-16 was applied to the typing of 42 human Brucella isolates obtained from 41 patients recovered from 2002 to 2006 at a tertiary-care center in Lebanon. All isolates were identified as Brucella melitensis by MLVA-16 and were found to be closely related to B. melitensis isolates from neighboring countries in the Middle East when their genotypes were queried against those in the web-based Brucella2007 MLVA database (http://mlva.u-psud.fr/). Panel 2B, which comprised the most variable loci, displayed a very high discriminatory power, while panels 1 and 2A showed limited diversity. The most frequent genotype comprised seven isolates obtained over 7 weeks in 2002, demonstrating an outbreak from a common source. Two isolates obtained from one patient 5 months apart comprised another genotype, indicating relapsing disease. These findings confirm that MLVA-16 has a good discriminatory power for species determination, typing of B. melitensis isolates, and inferring their geographical origin. Abbreviated panel 2B could be used as a short-term epidemiological tool in a small region of endemicity.

* Corresponding author. Mailing address for Mireille M. Kattar: Department of Pathology and Laboratory Medicine, American University of Beirut Medical Center, Cairo Street, Beirut, Lebanon. Phone: 961-1-374374, ext. 5175. Fax: 961-1-370845. E-mail: mireillekattar{at}gmail.com. Mailing address for Gilles Vergnaud: Université Paris-Sud 11, CNRS, UMR8621, Institut de Génétique et Microbiologie, Orsay 91405, France. Phone: 33-1-69156208. Fax: 33-1-69156678. E-mail: gilles.vergnaud{at}u-psud.fr

{triangledown} Published ahead of print on 15 October 2008.

{dagger} Supplemental material for this article may be found at http://jcm.asm.org/.

Journal of Clinical Microbiology, December 2008, p. 3935-3940, Vol. 46, No. 12
0095-1137/08/$08.00+0     doi:10.1128/JCM.00464-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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