Development of an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of bovine paratuberculosis
Por:
Covarrubias, AGM, Flores, MAS, Ruiz, CCG, Humara, LDF, Lopez, DC, Aparicio, ED, Andrade, LH, Ochoa, MAB
Publicada:
1 ene 2012
Resumen:
The objective of this study was to standardize and develop an ELISA with protoplasmatic antigen from a Mycobacterium avium paratuberculosis (Map) 3065 strain obtained from sheep, to be used for the diagnosis of bovine paratuberculosis. The antigen was set on plates at 10, 5, 2.5 and 1.25 mu g/100 mu l concentrations and 1:20, 1:40, 1:80 and 1:160 dilutions of positive and negative control sera were tested. The cutoff point was established with a confidence interval of 95% and two standard deviations. Four-hundred and ninety-one bovine sera were used to establish sensitivity and specificity of the ELISA; likewise feces samples were collected for bacteriological isolation and the nested polymerase chain reaction test (PCR) was applied for Map detection. Results of the three tests were analyzed by kappa test to determine association or concordance. Antigen and sera concentration that gave the best difference between controls was 1.25 mu g/100 mu l antigen and 1:160 serum dilution, cutoff
Filiaciones:
Covarrubias, AGM:
Univ Nacl Autonoma Mexico, Fac Med Vet & Zootecnia, Mexico City 04510, DF, Mexico
Ochoa, MAB:
Univ Nacl Autonoma Mexico, Fac Med Vet & Zootecnia, Mexico City 04510, DF, Mexico
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