Patent ID: 7303891

Claim:
A method for detecting antibodies against Lawsonia intracellularis in a swine serum sample comprising the steps of: a) adding about 0.1 ml/well of monoclonal antibody 110:9 produced by hybridoma cell line 110:9 deposited with the ECACC under the Accession No. 04092204 to each well of a standard ninety-six (96) well micro-titer plate wherein said monoclonal antibody has been diluted with coating buffer; b) incubating said micro-titer plate for eighteen (18) hours at about 4° C.; c) washing said incubated micro-titer plate three (3) times with phosphate buffered saline (PBS); d) adding about 0.1 ml/well of Lawsonia intracellularis antigen to each well of the washed micro-titer plate wherein said antigen has been diluted with buffer; e) incubating said micro-titer plate for 1 to 2 hours at 37° C.; f) washing three (3) times with wash buffer and/or directly adding 0.25 ml of blocking buffer to each well of the micro-titer plate; g) incubating said micro-titer plate for one (1) to two (2) hours at 37° C.; h) washing said micro-titer plate three (3) times with wash buffer; i) adding 0.1 ml of the swine serum to be tested to each well of the washed micro-titer plate wherein the swine serum has been diluted in blocking buffer; Alternatively, adding the swine serum to be tested to the first wells of the micro-titer plate and making serial 1:2 dilutions across the plate; j) incubating the resulting micro-titer plate for one (1) hour at 37° C.; k) washing said micro-titer plate three (3) to five (5) times with wash buffer; l) diluting a conjugated anti- L. intracellularis specific antibody in blocking buffer and adding 0.1 ml to each well of said micro-titer plate; m) incubating said micro-titer plate for one (1) hour at 37° C.; n) washing said micro-titer plate three (3) to five (5) times with wash buffer; o) adding substrate to each well of said washed micro-titer plate; p) measuring the absorbance of light in the wells of said micro-titer plate with a spectrophotometer.