Quick diagnosis of tuberculosis in cattle is critical to decide whether or not to quarantine or depopulate a herd. Currently, decisions are based on culture, which takes between four and eight weeks to accomplish. Therefore, the purpose of this study was to evaluate relative sensitivity and specificity of two PCR's (MPB70 and IS6110) and spoligotyping as quick diagnostic tests for cattle tuberculosis in fresh tissue. Fifty tissue samples with TB-suspicious lesions from a herd with 25 % prevalence of the disease and fifty tissue samples with no lesions from a TB-free herd were used in the study. Samples were split into two sets and each set was blind analyzed in two different laboratories under the same protocol. The three tests were used to diagnose tuberculosis in all samples using macerated tissue just before culturing. Relative sensitivity and specificity of all tests were estimated using presence/absence of lesion, histopathology and culture results as gold standards. MPB70 nested PCR showed consistently higher sensitivity (85 to 91 %) and specificity (77 to 86 %) in one of the laboratories with all gold standards; however, inter-laboratory differences occurred, in one of the laboratories sensitivity was from 89 to 91 % and specificity from 57 to 63 %. IS6110 nested PCR and spoligotyping behaved poorly. Relative sensitivity and specificity for MPB70 nested PCR suggest that this test could be useful as a complementary test for quick diagnosis of bovine TB in fresh tissue.

El diagnóstico rápido de tuberculosis en ganado es crítico para decidir si un hato debe someterse a cuarentena de manera definitiva o despoblarse. El objetivo del presente trabajo fue evaluar la sensibilidad y la especificidad relativa de dos PCR's (MPB70 y IS6110) y spoligotyping como pruebas rápidas de diagnóstico de tuberculosis bovina en tejido fresco. Se usaron 50 muestras de tejido con lesión, provenientes de un hato con 25 % de prevalencia de la enfermedad y 50 muestras de tejido sin lesión, provenientes de un hato libre de la enfermedad. Las muestras se dividieron en dos partes y cada parte analizada en ciego por dos laboratorios diferentes con un mismo protocolo. Las tres pruebas fueron utilizadas para el diagnóstico en tejido macerado tomado inmediatamente antes del cultivo. La sensibilidad y la especificidad relativa de las pruebas diagnósticas se determinaron usando el resultado de presencia/ ausencia de lesiones, histopatología y cultivo como "pruebas de oro." La PCR MPB70 demostró de manera consistente mayor sensibilidad (85 a 91 %) y especificidad (77 a 86 %) en uno de los laboratorios con todas las pruebas de oro. Sin embargo, se observaron diferencias inter-laboratorio: en el segundo laboratorio la sensibilidad fue de 89 a 91 %, pero la especificidad fue baja (57 a 63 %). La PCR IS6110 anidada y spoligotyping tuvieron un pobre comportamiento. La sensibilidad y la especificidad relativa para la PCR MPB70 sugiere que esta prueba puede ser de utilidad como prueba complementaria para el diagnóstico rápido.

OBJECTIVE: The purpose of the study was to determine the role of bovine TB in cases of human TB. MATERIAL AND METHODS: Two-hundred and fifty-five samples from symptomatic patients were included in the study. All samples were cultured in Stonebrink and Lowënstein-Jensen media and analyzed using a nested PCRMPB70. The molecular analysis was performed by spoligotyping. RESULTS: From 255 samples, 74 were PCR-positive and 20 were culture-positive. From 94 samples positive to PCR or to isolation, 66 (70%) showed a spoligotype compatible with M. tuberculosis, and 13 (13.8%) with M. bovis. Four fingerprints of M. bovis from humans were identical to the fingerprints of M. bovis from cattle in the same region. CONCLUSIONS: Our study shows that M. bovis plays an important role in the epidemiology of TB in humans and that TB in cattle represents a risk to public health.

OBJETIVO: Determinar el papel de la tuberculosis bovina en la tuberculosis humana. MATERIAL Y MÉTODOS: Se analizaron 255 muestras de pacientes sintomáticos, sembradas en medios de Stonebrink y Löwenstein-Jensen y analizadas por PCRMPB70 anidada y luego por spoligotyping. RESULTADOS: De las 255 muestras, 74 fueron positivas a la PCR y 20 al aislamiento: de las primeras, 58 (78%) mostraron espoligotipo de M. tuberculosis y 5 (6.7%) de M. bovis; de las segundas, 8 (47%) revelaron espoligotipo de M. tuberculosis y 8 (47%) de M. bovis. De las 94 muestras positivas al aislamiento o PCR, 66 (70%) correspondieron a M. tuberculosis y 13 (13.8%) a M. bovis. Los patrones moleculares de cuatro muestras de M. bovis de seres humanos fueron idénticos a los de las cepas de M. bovis de ganado. CONCLUSIONES: Se demuestra que M. bovis juega un papel importante en la epidemiología de la tuberculosis humana y representa un riesgo para la salud pública.

OBJECTIVE: The purpose of the study was to determine the role of bovine TB in cases of human TB. MATERIAL AND METHODS: Two-hundred and fifty-five samples from symptomatic patients were included in the study. All samples were cultured in Stonebrink and Lowënstein-Jensen media and analyzed using a nested PCRMPB70. The molecular analysis was performed by spoligotyping. RESULTS: From 255 samples, 74 were PCR-positive and 20 were culture-positive. From 94 samples positive to PCR or to isolation, 66 (70%) showed a spoligotype compatible with M. tuberculosis, and 13 (13.8%) with M. bovis. Four fingerprints of M. bovis from humans were identical to the fingerprints of M. bovis from cattle in the same region. CONCLUSIONS: Our study shows that M. bovis plays an important role in the epidemiology of TB in humans and that TB in cattle represents a risk to public health.

OBJETIVO: Determinar el papel de la tuberculosis bovina en la tuberculosis humana. MATERIAL Y MÉTODOS: Se analizaron 255 muestras de pacientes sintomáticos, sembradas en medios de Stonebrink y Löwenstein-Jensen y analizadas por PCRMPB70 anidada y luego por spoligotyping. RESULTADOS: De las 255 muestras, 74 fueron positivas a la PCR y 20 al aislamiento: de las primeras, 58 (78%) mostraron espoligotipo de M. tuberculosis y 5 (6.7%) de M. bovis; de las segundas, 8 (47%) revelaron espoligotipo de M. tuberculosis y 8 (47%) de M. bovis. De las 94 muestras positivas al aislamiento o PCR, 66 (70%) correspondieron a M. tuberculosis y 13 (13.8%) a M. bovis. Los patrones moleculares de cuatro muestras de M. bovis de seres humanos fueron idénticos a los de las cepas de M. bovis de ganado. CONCLUSIONES: Se demuestra que M. bovis juega un papel importante en la epidemiología de la tuberculosis humana y representa un riesgo para la salud pública.

Bovine tuberculosis, caused by Mycobacterium bovis, is a disease of socio-economic and public health importance and of significance to international trade regulation. Allelic variants of several genes have been implicated in the genetic susceptibility to tuberculosis in some human populations, but little is known in cattle. We surveyed 34 European, 18 Asian, 20 Creole and 23 hybrid bovines for polymorphisms of the bovine solute carrier family 11 a1(Slc11a1) gene, formerly known as natural resistance associated macrophage protein (Nramp1), gene by typing the cattle using two microsatellite loci closely linked to this gene. The microsatellites used were 311-22, located at the 3' untranslated region (3' UTR) of the Slc11a1 gene, and ARO28 situated about 0.6 cM upstream of the same gene Based on allele size in base pairs (bp) we determined five 311-223 locus variants (221, 223, 225, 227 and 229 bp) and 12 ARO28 loci. There was marked diversity and a very high level of heterozygosity in most of the cattle surveyed except the Europeans bovines and especially Holsteins in relation to the 3' UTR microsatellite locus.