Spoligotyping and variable number tandem repeat analysis of Mycobacterium bovis isolates from cattle in Brazil.
We performed spoligotyping and 12-mycobacterial interspersed repetitive unit-variable number tandem repeats (MIRU-VNTRs) typing to characterise Mycobacterium bovis isolates collected from tissue samples of bovines with lesions suggestive for tuberculosis during slaughter inspection procedures in abattoirs in Brazil. High-quality genotypes were obtained with both procedures for 61 isolates that were obtained from 185 bovine tissue samples and all of these isolates were identified as M. bovis by conventional identification procedures. On the basis of the spoligotyping, 53 isolates were grouped into nine clusters and the remaining eight isolates were unique types, resulting in 17 spoligotypes. The majority of the Brazilian M. bovis isolates displayed spoligotype patterns that have been previously observed in strains isolated from cattle in other countries. MIRU-VNTR typing produced 16 distinct genotypes, with 53 isolates forming eight of the groups, and individual isolates with unique VNTR profiles forming the remaining eight groups. The allelic diversity of each VNTR locus was calculated and only two of the 12-MIRU-VNTR loci presented scores with either a moderate (0.4, MIRU16) or high (0.6, MIRU26) discriminatory index (h). Both typing methods produced similar discriminatory indexes (spoligotyping h = 0.85; MIRU-VNTR h = 0.86) and the combination of the two methods increased the h value to 0.94, resulting in 29 distinct patterns. These results confirm that spoligotyping and VNTR analysis are valuable tools for studying the molecular epidemiology of M. bovis infections in Brazil.
Instituto René Rachou-Fiocruz, Belo Horizonte, MG, Brasil.
Patrícia Martins
PM
Parreiras
Giovanna Ivo
GI
Andrade
Telma de Figueiredo do
Tde F
Nascimento
Maraníbia Cardoso
MC
Oelemann
Harrison Magdinier
HM
Gomes
Andrea Padilha de
AP
Alencar
Ronnie Antunes de
RA
Assis
Pedro Moacyr Pinto Coelho
PM
Mota
Márcia Aparecida da Silva
MA
Pereira
Francisco Carlos Faria
FC
Lobato
Andrey Pereira
AP
Lage
Philip Noel
PN
Suffys
D000483:Alleles; D000818:Animals; D015373:Bacterial Typing Techniques / Q000379:methods*; D002417:Cattle; D004269:DNA, Bacterial / Q000235:genetics; D014644:Genetic Variation / Q000235:genetics*; D005838:Genotype; D009163:Mycobacterium bovis / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D020080:Tandem Repeat Sequences / Q000235:genetics*
D004269:DNA, Bacterial