02453naa a2200301 a 450000100080000000500110000800800410001910000180006024500820007826000090016030000200016952015550018965000160174465000100176065000240177065000200179470000180181470000250183270000200185770000200187770000200189770000260191770000170194370000220196070000180198270000150200077301360201516633702010-04-07       2009    bl uuuu     u00u1 u    #d1 aFARIAS, T. A.  aELISA based on recombinant MPB70 to detect antibodies to Mycobacterium bovis.  c2009  a1 p.c1 CD-ROM.  aBovine tuberculosis is an important infectious disease, caused by Mycobacterium bovis. The gene that encodes for MPB70 protein is present in the genome of M. bovis and has high specificity for the M. tuberculosis complex, representing a potential antigen for the diagnosis of bovine tuberculosis. The objective of this work was to develop an ELISA using recombinant protein MPB70 of M. bovis in order to detect antibodies against these bacteria in cattle. The gene mpb70 was amplified by PCR and cloned into plasmid pGEM-T Easy. After digestion with the Eco RI enzyme, the gene was subcloned into the expression plasmid pGEX-3X. The expression of the gene was verified by SDS-PAGE and Western blot with anti-glutathione-S-transferase. After standardization of the ELISA, serum samples of positive (53), negative (37) or inconclusive (47) cattle in the tuberculin test were tested. In SDS-PAGE, a recombinant protein of approximately 48 kDa was detected, which corresponds to the estimated molecular weight of GST/MPB70. In Western blot, recombinant MPB70, fused to GST, was recognized by anti-GST antibody, confirming the expression of the gene. For the ELISA with recombinant MPB70, the sensitivity and specificity found were 88.7% and 94.6%, respectively. The use of a serological test such as ELISA with recombinant MPB70, together with the cellular tests, may solve some of the problems found in the control programs of bovine tuberculosis, like inconclusive results and the lack of detection of anergic animals in advanced stages of the disease.  aDiagnostico  aElisa  aMycobacterium Bovis  aSanidade Animal1 aARAUJO, F. R.1 aOSÓRIO, A. L. A. R.1 aJORGE, K. S. G.1 aRAMOS, C. A. N.1 aSOUZA, I. I. F.1 aOLIVEIRA, R. H. M. de1 aAZAMBUJA, A.1 aROSINHA, G. M. S.1 aSOARES, C. O.1 aELISEI, C.  tIn: SIMPÓSIO EMBRAPA LABEX DE SANIDADE ANIMAL, 1., 2009, Campo Grande, MS. Anais... Campo Grande, MS: Embrapa Gado de Corte, 2009.