TH45733B - Vaccines and reagents for the diagnosis of pig zircoviruses - Google Patents

Abstract

DC60 (03/02/55) This invention relates to novel strains of porcine circovirus isolated from lung or nerve ganglion samples obtained from farms experiencing post-weaning multisystematic weaning syndrome (PMWS). It relates to purified preparations of these strains, attenuated or inactivated vaccines, live recombinant vaccines, plasmid vaccines and commonly used vaccine subunits, as well as reagents and methods for diagnosis. It also relates to DNA fragments that can be used for the production of subunits in characterized vectors under in vitro testing or in sequences that will be inserted into viruses or plasmids in characterized vectors under in vitro testing. This invention relates to novel strains of porcine circovirus isolated from lung or nerve ganglion samples obtained from farms experiencing post-weaning multisystematic weaning syndrome (PMWS). It relates to purified preparations of these strains, attenuated or inactivated vaccines, live recombinant vaccines, plasmid vaccines and commonly used vaccine subunits. As well as reagents and methods for diagnosis, it also involves DNA fragments that can be used to produce subunits in characterizing vectors under in vitro conditions or in sequences that will be embedded into viruses or plasmids in characterizing vectors under in vivo conditions.

Claims (41)

1. Vaccines prepared from antigen preparations collected from in vitro cell cultures infected with selected isolated porcine circovirus type II from those deposited in the ECACC under reference sections V97100219, V97100218, V97100217, V98011608, V98011609, or clear fractions or culture extracts containing porcine circovirus type II, collected from such isolated in vitro cell cultures infected with porcine circovirus type II as antigen. 2. Purified porcine circovirus preparations selected from a pool of preparations deposited with ECACC under the following references: Access No. V97100219, Access No. V97100218, Access No. V97100217, Access No. V98011608, Access No. V98011609. 3. Preparation of porcine circoviruses isolated from cells in in vitro. These cells were infected with porcine circoviruses that could be isolated from physiological or tissue samples, particularly from lesions in pigs suffering from PMWS syndrome. 4. Preparation of porcine circovirus pursuant to claim 3, produced and isolated from porcine kidney cell lines. 5. Preparation according to claim 4, produced and isolated from undefiled PK/15 cells using PCV. 6. An extract or aqueous fraction of cell cultures collected from in vitro cell cultures of cells infected with the circovirus described in Reputation 1. 7. Preparation of antigens collected from in vitro cultured cells infected with circovirus according to claim 1. 8. A vaccine containing an antigen preparation pursuant to claim No. 7, which includes porcine circovirus as the antigen. 9. The vaccine under claim 8 is characterized by the presence of a fully attenuated, live viral antigen in a veterinary-accepted carrier or diluent, and may be supplemented with a veterinary-accepted adjuvant, as well as with a freeze-drying stabilizer. 10. The vaccines under claim 9 exhibit an inactivated antigen and are supplemented with a veterinarily acceptable carrier or diluent, and may be further supplemented with a veterinarily acceptable adjuvant. 11. The vaccine under claim 8 is characterized by the inclusion of antigens of several porcine circoviruses. 12. The vaccine under claim 8 is characterized by the inclusion of at least one additional valency corresponding to another swine pathogen. 13. A DNA fragment containing a sequence selected from a group composed of sequences defined by the reference segments of SEQ ID NO:1 SEQ ID NO:2 SEQ ID NO:3 SEQ ID NO:4 SEQ ID NO:6 14. DNA fragments containing ORFs selected from a group containing ORFs 1 through 13. 15. The DNA fragment under claim 14 exhibits a characteristic where it has an ORF selected from a group comprising ORFs 4, 7, 10, and 13. 16. A polypeptide encoded by a DNA fragment according to claim 13. 17. A polypeptide encoded by a DNA fragment according to claim 14. 18. The expression of a vector in an in vitro environment involves the insertion of a DNA sequence or fragment under patent 13 into its genome so that it can express itself in an in vitro. 19. The expression of a vector in an in vitro environment consisting of the insertion of a DNA sequence or fragment under claim 14 into its genome so that it can express its traits in an in vitro environment. 20. The vector characterized under claim 18 is characterized by its selection from E. coli and Badulovirus. 21. Vector-generated polypeptides exhibit characteristics pursuant to claim 18. 22. Vector-generated polypeptides exhibit characteristics pursuant to claim 19. 23. The vaccine subunit shall consist of at least one polypeptide as per claim 21 in a veterinary acceptable modifier or carrier and shall be supplemented with a veterinary acceptable adjuvant. 24. A subunit vaccine containing at least one polypeptide as per claim 21 in a diluent or carrier acceptable from a veterinary perspective, and may be optionally supplemented with an adjuvant acceptable from a veterinary perspective. 25. The bodily expression of a vector consisting of the insertion of a DNA fragment under claim 13 into its genome so that it can exhibit bodily expression. 26. The in vivo expression of a vector consisting of a DNA fragment under claim 14 inserted into its genome to enable in vivo expression. 27. A characterization vector pursuant to claim 25 is selected from a live virus capable of replication in pigs without causing pathogenicity to this animal and a plasmid. 28. A characterization vector pursuant to claim 26 is characterized by selection from a live virus capable of replication in pigs without causing disease in this animal and a plasmid. 29. Characterization vectors under Recourse 27 are represented by a virus vector selected from swine herpesviruses, such as Aujeszky's disease virus, swine adenovirus, specific pod viruses, vaginia, avipoxvirus, canary pod virus, or swine pod virus. 30. Characterization vectors under Receipt 28 are represented by a virus vector selected from swine herpesviruses, such as Aujeszky's disease virus, swine adenovirus, poxviruses, particularly vaceniavirus, avipoxvirus, canarypoxvirus, and swinepoxvirus. 31. Live attenuated vaccines or plasmids exhibiting characteristics as defined by Patent 25 in a veterinaryly accepted carrier or dilutor. 32. Live attenuated vaccines or plasmids exhibiting characteristics as defined in Patent 26 in a veterinaryly accepted carrier or dilutor. 33. A probe or primer containing all or part of the sequence as described in Claim 13. 34. A probe or primer containing all or part of the sequence as described in Claim 14. 35. Polyclonal or monoclonal antibodies prepared from circoviruses according to Relieve 1, or from polypeptides encoded by DNA fragments containing sequences selected from the group containing SEQ ID NOS. 1,2,3,4, and 6, or from polypeptides characterized by vectors containing sequences selected from the group containing SEQ ID NOS. 1,2,3,4, and 6, or from polypeptides characterized by vectors containing DNA including ORFs selected from the group containing ORFs 1 through 13. 36. Method for detecting porcine circovirus in physiological fluid or tissue samples from pigs. Testing is performed to detect the presence of antigens by identifying the antigens themselves or antibodies produced against these antigens. 37. A vaccine consisting of a clear cell culture fraction or an extract as per claim 6 containing porcine circovirus as an antigen. 38. A vaccine under any of the claims of claims 23, 24, 31, or 32 is characterized by the inclusion of antigens of several porcine circoviruses. 39. A vaccine under any of the claims of Claim 23, 24, 31, or 32 is characterized by the inclusion of at least one valency corresponding to another swine pathogen. 40. Vaccines under claims 11, 12, 38, or 39 are characterized by the inclusion of at least one selected valency from a group comprising PRRS, Mycoplasma hyopneumoniae, E. coli, Atrophic Rhinitis, pseudorabies, swine cholera, and swine influenza. 41. The vaccine under claim 40 is characterized by the inclusion of at least one valency selected from the group comprising PRRS, Mycoplasma hyopneumoniae (Clause 41, page 4, figure 7).