Oral communication in Workshop "VNTR/MIRU and DVR-Spoligotyping for Mycobacterium bovis typing"

March 25^th, 2009

Rodriguez-Campos S., Castellanos E., de Juan L., Bezos J., Gallardo F., Moya N., Alvarez J., Alvarez N., Alende T., Gutierrez A., Lozano F., Mateos A. and Romero B.

Among Spanish Mycobacterium (M.) bovis isolates the most prevalent spoligotype pattern is SB0121, which contains all M. bovis spacers except number 21. This spoligotype accounts for 27.9% of the M. bovis isolates and has been found throughout the country over the last 16 years. Furthermore, this spoligotype, which represents almost 30% of the cattle isolates, is also detected in goats and wildlife (red deer, fallow deer and wild boar). Other Mediterranean countries such as France, Portugal and Italy also described the isolation of SB0121, as well as reports from South Africa and South America. In contrast, no SB0121 isolates were reported from the British Isles. In order to determine whether the high prevalence of this spoligotype can be due to incapacity of the DVR-spoligotyping to further discriminate these strains, we analysed a representative panel of SB0121 isolates (n=100) by MIRU-VNTR typing. A group of six loci was used to subtype the panel: ETRA (VNTR2165), ETRB (VNTR2461), ETRD (VNTR580), QUB11b
(VNTR2163b), QUB3232 (VNTR3232, short fragment) and QUB26 (VNTR4052). Five of the loci were included in the Multilocus Variable Number Tandem Repeat Genotyping of M. bovis Ring Trial VENoMYC 2006, and QUB26 was chosen because by our experience it provides a better discrimination than QUB11a (VNTR2163a). A differentiation into 55 profiles was achieved, with a discriminatory power (D) of 0.97. The three most discriminative loci in decreasing order were: VNTR3232, ETRA and ETRB. This set of loci resulted highly discriminative for the Spanish M. bovis SB0121 spoligotype and therefore indicates the lack of discriminatory power of DVR- spoligotyping in this particular case

	Servicio de Micobacterias (MYC). Centro de Vigilancia Sanitaria Veterinaria (VISAVET). Universidad Complutense (UCM).
	Departamento de Sanidad Animal. Facultad de Veterinaria. Universidad Complutense (UCM).