MIRU-VNTR typing adds discriminatory value to groups of Mycobacterium bovis and Mycobacterium caprae strains defined by spoligotyping

• Published in: Veterinary microbiology Vol. 143; no. 2; pp. 299 - 306
• Main Authors: Duarte, E.L., Domingos, M., Amado, A., Cunha, M.V., Botelho, A.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 14.07.2010; Amsterdam; New York: Elsevier; Elsevier
• Subjects: accuracy; Animals; Animals, Wild; Bacterial Typing Techniques - methods; Bacteriology; Biological and medical sciences; Bovine tuberculosis; Cattle; Cluster Analysis; Deer; disease diagnosis; disease reservoirs; disease transmission; Fundamental and applied biological sciences. Psychology; genetic variation; Genotype; loci; Microbiology; minisatellite repeats; MIRU-VNTR typing; Miscellaneous; molecular epidemiology; molecular sequence data; Mycobacterial Interspersed Repetitive Units-Variable Number Tandem Repeats; Mycobacterium - classification; Mycobacterium - genetics; Mycobacterium bovis; Mycobacterium caprae; new methods; pathogen identification; pathotypes; Phylogeny; sequence analysis; Spoligotyping; strains; Sus scrofa; Swine; validity; wildlife; wildlife diseases; Portugal; Spoligotyping; MIRU-VNTR typing; Mycobacterium bovis; Mycobacterium caprae; Bovine tuberculosis; Typing; Bovine; Genotype; Strain; Vertebrata; Mammalia; Mycobacteriales; Minisatellite DNA; Mycobacteriaceae; Bacteria; Actinomycetes; Artiodactyla; Veterinary; Ungulata
• ISSN: 0378-1135; 1873-2542
• DOI: 10.1016/j.vetmic.2009.11.027

The value of Mycobacterial Interspersed Repetitive Units-Variable Number Tandem Repeats (MIRU-VNTR) as a genotyping technique for Mycobacterium bovis and Mycobacterium caprae, has been confirmed in different countries and epidemiological scenarios. However, a standardized panel of loci has not yet been adopted for these species, since allelic diversity of each locus differs among countries. To determine the most discriminatory loci, a panel of 181 M. bovis and M. caprae strains representing 12 spoligotypes was created. The panel included strains from the three predominant spoligotypes previously isolated in Portugal; strains from spoligotyping group SB0140, prevalent in the British Isles but also detected in Portugal; strains from spoligotypes common to cattle and wildlife species and strains from the M. caprae spoligotyping group SB0157. MIRU-VNTR analysis of these strains, targeting 8 selected loci, produced 87 different profiles ( h = 0.99), being VNTR3232, QUB11a, ETR-B and ETR-A the most discriminatory loci ( h = 0.96). A single M. bovis spoligotyping group could be differentiated – up to 44 MIRU-VNTR profiles. These results emphasize the high genotype diversity of Portuguese isolates compared with other countries. MIRU-VNTR typing was superior to spoligotyping for identifying multi-genotype infected herds and the combination of the two genotyping methods by a hierarchical approach confirmed the genetic relatedness of M. bovis isolates between cattle and wildlife.