Vaccine diagnostics
View Patent ↗The present invention relates to improved diagnostic methods and kits for differentiating between (a) animals administered a chimeric pestivirus, and (b) animals infected with a wild-type bovine viral diarrhea virus (BVDV) or immunized with a conventional BVDV vaccine.
1. A method of detecting the presence of an antibody that specifically binds to a bovine viral diarrhea virus (BVDV) E rns protein, said method comprising the steps of:
a) incubating a serum sample from an animal with pronghorn pestivirus E rns protein or a fragment thereof to which antibodies that cross-react with BVDV E rns bind, wherein said sample is incubated either prior to or concurrently with step b);
b) adding said sample from a) to an assay plate comprising bound BVDV E rns protein; and
c) detecting in said sample the presence of said antibody, wherein the detecting comprises adding a labeled BVDV E rns -specific monoclonal antibody to the assay plate, wherein the labeled antibody will bind to the bound BVDV E rns protein.
2. The method of claim 1 , wherein said antibody to be detected specifically binds to at least one epitope present in a BVDV E rns protein, but not present in a pronghorn pestivirus E rns protein.
3. The method of claim 1 , wherein the presence of said antibody indicates that the animal has either been infected with wild-type BVDV or immunized with a conventional BVDV vaccine.
4. The method of claim 1 , wherein the absence of said antibody indicates that the animal has not been 1) infected with wild-type BVDV, or 2) immunized with a conventional BVDV vaccine, or has been immunized with a chimeric pestivirus expressing a pronghorn pestivirus E rns protein.
5. The method of claim 1 , wherein the animal is an animal that is susceptible to BVDV infections.
6. The method of claim 5 , wherein the animal is a bovine, ovine, caprine, or porcine species.
7. The method of claim 6 , wherein the animal is a bovine.
8. A diagnostic kit useful for carrying out the method of claim 1 , wherein said kit comprises components that facilitate the detection of the presence in the serum sample from an animal of an antibody to at least one BVDV E rns epitope not present in a pronghorn pestivirus E rns protein, wherein said components comprise pronghorn pestivirus E rns protein or a fragment thereof to which antibodies that cross-react with BVDV E rns bind, an assay plate comprising bound BVDV E rns protein, and a labeled BVDV E rns -specific monoclonal antibody.
9. The diagnostic kit of claim 8 , wherein said labeled BVDV E rns -specific monoclonal antibody is an antibody that specifically binds to an epitope present in wild-type BVDV or a conventional BVDV vaccine, but which epitope is not present in a pronghorn pestivirus E rns protein.
10. The diagnostic kit of claim 8 , wherein said kit comprises reagents for carrying out an assay selected from the group consisting of: an enzyme-linked immunosorbent assay (ELISA), a lateral flow assay, Western blotting, PCR, radioimmunoassay, solid phase radioimmunoassay, electrochemiluminescent assay, immunoblotting, immunoprecipitation, and immunostaining.
11. The method of claim 1 , further comprising testing for the presence of (i) a BVDV protein that is not E rns or (ii) an antibody that specifically binds a BVDV protein that specifically binds a BVDV protein that is not E rns .
12. The method of claim 11 , wherein the sample is tested for the presence of an antibody that specifically binds to a BVDV protein, further wherein said protein is selected from the group consisting of p80 and E2.