Ileitis diagnostic assay
Improved immunoassays for the protection of antibodies against Lawsonia intracellularis are provided which permit rapid, easy detection of low concentrations of anti- Lawsonia antibodies in animal-derived specimens. The preferred assay is an ELISA assay employing an antigenic extract of L. intracellularis lipopolysaccharide.
1. An immunoassay for detecting the presence of antibodies against Lawsonia intracellularis in an animal-derived specimen comprising the steps of contacting said specimen with an effective amount of an antigen comprising at least a portion of an isolated lipopolysaccharide of Lawsonia intracellularis , said portion being of sufficient size to form antibody-antigen complexes with Lawsonia intracellularis LPS antibodies and having an endotoxicity of about 3-75 EU/ml, causing the formation of complexes between said antigen and said antibodies, and determining the presence of said complexes, wherein the presence of said complexes indicates the presence of said antibodies.
2. The immunoassay of claim 1 , said specimen selected from the group consisting of animal-derived sera, colostrums, joint fluids, salivas, tissue homogenates, and feces.
3. The immunoassay of claim 1 , said animal selected from the group consisting of pigs, hamsters, blue fox, emus, deer, dogs, guinea pigs, horses, rhesus macaque monkeys, ostriches, rabbits and rats.
4. The immunoassay of claim 1 , said antibodies being selected from the group consisting of IgG, IgA and IgM antibodies.
5. The immunoassay of claim 1 , said immunoassay being an ELISA test.
6. The immunoassay of claim 5 , said antigen being recombinantly derived.
7. The immunoassay of claim 5 , said ELISA test including the steps of immobilizing said antigen on a surface, contacting said specimen with said immobilized antigen, causing a complex between antibodies in said specimen and said antigen, and detecting said complexes.
8. The immunoassay of claim 1 , said lipopolysaccharide having a molecular weight of from about 15-25 kDa.
9. The immunoassay of claim 8 , said molecular weight being from about 18-20 kDa.
10. The immunoassay of claim 1 , said antigen capable of inducing an immune response in said animal upon administration of an effective amount of the antigen to the animal.
11. The immunoassay of claim 1 , said endotoxicity being from about 25-40 EU/ml.
12. The immunoassay of claim 1 , said antigen comprising an isolated lipopolysaccharide extract from the strain deposited as ATCC Accession No. PTA-4927.
13. An antigen for detecting the presence of antibodies against Lawsonia intracellularis , said antigen comprising at least a portion of an isolated Lawsonia intracellularis lipopolysaccharide, said portion being sufficient size to form antibody-antigen complexes with Lawsonia intracellularis LPS antibodies and having an endotoxicity of about 3-75 EU/ml.
14. The antigen of claim 13 , said lipopolysaccharide having a molecular weight of from about 15-25 kDa.
15. The antigen of claim 14 , said molecular weight being from about 18-20 kDa.
16. The antigen of claim 14 , said antigen capable of inducing an immune response in said animal upon administration of an effective amount of the antigen to the animal.
17. The antigen of claim 14 , said lipopolysaccharide being recombinantly derived.
18. The antigen of claim 14 , said endotoxicity being from about 25-40 EU/ml.
19. The antigen of claim 13 , said antigen comprising an isolated lipopolysaccharide extract from the strain deposited as ATCC Accession No. PTA-4927.
20. In an enzyme-linked immunoassay for detecting the presence of antibodies against Lawsonia intracellularis in an animal-derived specimen, the improvement which comprises employing an isolated lipopolysaccharide of Lawsonia intracellularis having an endotoxicity of about 3-75 EU/ml, as an antigen in said immunoassay.
21. The immunoassay of claim 20 , said specimen selected from the group consisting of animal-derived sera, colostrums, joint fluids, salivas, tissue homogenates, and feces.
22. The immunoassay of claim 20 , said animal selected from the group consisting of pigs, hamsters, blue fox, emus, deer, dogs, guinea pigs, horses, rhesus macaque monkeys, ostriches, rabbits and rats.
23. The immunoassay of claim 20 , said antibodies being selected from the group consisting of IgG, IgA and IgM antibodies.
24. The immunoassay of claim 20 , said ELISA test being an indirect-ELISA test.
25. The immunoassay of claim 24 , said ELISA test including the steps of immobilizing said antigen on a surface, contacting said specimen with said immobilized antigen, causing a complex between antibodies in said specimen and said antigen, and detecting said complexes.
26. The immunoassay of claim 20 , said lipopolysaccharide having a molecular weight of from about 15-25 kDa.
27. The immunoassay of claim 26 , said molecular weight being from about 18-20 kDa.
28. The immunoassay of claim 20 , said antigen capable of inducing an immune response in said animal upon administration of an effective amount of the antigen to the animal.
29. The immunoassay of claim 20 , said endotoxicity being from about 25-40 EU/ml.
30. The immunoassay of claim 20 , said antigen comprising an isolated lipopolysaccharide extract from the strain deposited as ATCC Accession No. PTA-4927.