Evaluation and selection of tandem repeat loci for a Brucella MLVA typing assay

• DOI: 10.1186/1471-2180-6-9
• Publication Date: 2006-02-10T07:21:46Z
• Authors: Philippe Le Flèche; Isabelle Jacques; Maggy Grayon; Sascha Al Dahouk; Patrick Bouchon; France Denoeud; Karsten Nöckler; Heinrich Neubauer; Laurence A Guillo...; Gilles Vergnaud
• Keywords: Biovar; Minisatellite

Abstract Background The classification of Brucella into species and biovars relies on phenotypic characteristics sometimes raises difficulties in the interpretation results due to an absence standardization typing reagents. In addition, resolution this biotyping is moderate requires manipulation living agent. More efficient DNA-based methods are needed, work explores suitability multiple locus variable number tandem repeats analysis (MLVA) for both identification. Results Eighty repeat loci predicted be polymorphic by genome sequence three available sequences were tested polymorphism genotyping 21 strains (18 reference representing six 'classical' all as well 3 marine mammal currently recognized members two new species). MLVA data efficiently cluster expected according their biovar. For practical use, a subset 15 preserving clustering was selected applied 236 isolates. Using MLVA-15 assay, clusters generated correspond classical scheme spp. markers have been divided groups, one comprising 8 user-friendly minisatellite with good identification capability (panel 1) another complementary group 7 microsatellite higher discriminatory power 2). Conclusion assay can large collections automated or manual procedures, proposed complement, even substitute, methods. This facilitated fact that based non-infectious material (DNA) whereas procedure itself produced queried dedicated web service site.