VISAVET Health Surveillance Centre is the institution responsible for the database where national data regarding animal mycobacteriosis are registered since 1996 to the present days.
The database has an additional viewer for visualization of the geographic distribution of mycobacterial isolations according to searching criteria introduced by the user (such as year of isolation, host species from which the bacteria was cultured or spoligotypes involved).
Access to this database is available for Veterinary Services and Laboratories involved in the National Bovine Tuberculosis Eradication Program using the Veterinary Health Alert Network (RASVE) webpage of the Ministry of Agriculture, food and Environmental Affairs
The Direct Variable Repeat Spacer Oligonucleotide Typing technique or DVR-spoligotyping is based on polymorphism of the chromosomal DR locus, which consists of a variable number of direct repeats (DR) interspersed with nonrepetitive spacers. The spacers are amplified by polymerase chain reaction (PCR) and detected by hybridization of the biotin-labelled PCR product with a membrane containing oligonucleotides derived from spacer sequences that have previously been bound to a membrane. At the end of the protocol a profile is obtained characterized by the presence or absence of spacers. This technique is specific for bacterial species included in the M. tuberculosis complex (M. tuberculosis, M. bovis, M. caprae, M. africanum, M. pinnipedii and M. microti). Nowadays, the DVR-spoligotyping technique is the method of choice for epidemiological studies (wildlife and domestic transmission, animal movement, isolates specific of a geographic region, outbreaks, etc.)
References:
Aranaz A, Liébana E, Mateos A, Dominguez L, Vidal D, Domingo M, Gonzolez O, Rodriguez-Ferri EF, Bunschoten AE, Van Embden JD, Cousins D. Spacer oligonucleotide typing of Mycobacterium bovis strains from cattle and other animals: a tool for studying epidemiology of tuberculosis. J Clin. Microbiol. 34(11):2734-40. 1996.
Kamerbeek J., Schouls L, Kolk A, van Agterveld M, van Soolingen D, Kuijper S, Bunschoten A, Molhuizen H, Shaw R, Goyal M, van Embden J. Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology. J Clin. Microbiol. 35(4):907-14. 1997.
Mycobacterial Interspersed Repetitive Units -Variable Number Tandem Repeats (MIRU-VNTR) genotyping is a PCR-based technique that amplifies a selection of MIRU-VNTR loci. These loci consist of elements found as tandem repeats (from 40 to 120 base pair) and dispersed in intergenic regions in the genome of the members of the Mycobacterium tuberculosis complex . The number of repeated sequences of each locus is estimated by analysing the amplicon size after electrophoresis migration. The VNTR 3232, QUB 11a, QUB 11b, ETR-A, ETR-B and MIRU 26 loci, among others, have been proven highly discriminatory for M. bovis isolates. However, MIRU-VNTR analysis is not yet standardised in M. bovis and allelic diversity can vary among countries.
This method has become an important technique, since it allows high-throughput, discriminatory and reproducible analysis of the Mycobacterium tuberculosis complex strains. MIRU-VNTR has been proposed as an additional tool to DVR-spoligotyping for the genotyping of M. tuberculosis complex strains.
Currently the Health Surveillance Centre uses this technique in the epidemiological study of specific outbreaks.
References:
Frothingham R, Meeker-O'Connell WA. Genetic
diversity in the Mycobacterium tuberculosis complex based on variable numbers of tandem DNA repeats. Microbiology. 1998 May;144
(5):1189-96.
Aranaz A, Romero B, Montero N, Alvarez J, Bezos J, de Juan L, Mateos A, Dominguez L. Spoligotyping profile change caused by deletion of a
direct variable repeat in a Mycobacterium tuberculosis isogenic laboratory strain. J Clin Microbiol. 2004 Nov;42(11):5388-91.
Prodinger WM, Brandstätter A, Naumann L, Pacciarini M, Kubica T, Boschiroli ML, Aranaz A, Nagy G, Cvetnic Z, Ocepek M, Skrypnyk A,
Erler W, Niemann S, Pavlik I, Moser I. Characterization of
Mycobacterium caprae isolates from Europe by mycobacterial interspersed repetitive unit genotyping. J Clin Microbiol. 2005
Oct;43(10):4984-92.
Romero B, Aranaz A, de Juan L, Alvarez J, Bezos J, Mateos A, Gomez-Mampaso E, Dominguez L. Molecular epidemiology of multidrug-resistant
Mycobacterium bovis isolates with the same spoligotyping profile as isolates from animals. J Clin Microbiol. 2006 Sep;44(9):3405
-8.
Romero B, Aranaz A, Sandoval A, Alvarez J, de Juan L, Bezos J, Sanchez C, Galka M, Fernandez P, Mateos A, Dominguez L. Persistence and molecular
evolution of Mycobacterium bovis population from cattle and wildlife in Doñana National Park revealed by genotype variation. Vet
Microbiol. 2008 Nov 25;132(1-2):87-95
