Resumen
The present invention relates to the use of a new isolate of Neospora caninum for the development of diagnostic tests and for the manufacture of products for the treatment and prevention of the infection caused by Neospora. The use of a new isolate of Neospora caninum (filed in the Culture Collection of Algae and Protozoa , with access number CCAP 2051/1), and the extracts obtained therefrom, in the preparation of diagnostic tests and in the manufacture of products for the treatment and prevention of the infection caused by Neospora, is based on the ease with which the new isolate converts from the tachyzoite to the bradyzoite stage under in vitro conditions. Additionally and with the same applications, the polypeptides and the polynucleotides obtained from the new isolate and the immunoreactive antibodies developed against Neospora antigens are used.
Reivindicaciones
1. Parasite isolated from bovine nervous tissue, of the species Neospora caninum and called Nc-Spain 7, deposited in the Culture Collection of Algae and Protozoa with access number CCAP 2051 /1. 2. Use of biologically pure cell cultures infected by the isolate Nc-Spain 7 of claim 1 for obtaining tachyzoites, bradyzoites, polypeptides, antigens, antibodies, nucleic acids and other compounds of use in the preparation of products for the diagnosis, treatment and prevention of neosporosis. 3. Use of biologically pure cell cultures infected by the isolate Nc-Spain 7, according to claim 2, characterized by being cultures of vervet monkey kidney cells. 4. Use of the isolate Nc-Spain 7 of claim 1 in the diagnosis of the infection caused by Neospora. 5. Use of the isolate Nc-Spain 7 according to claim 4, characterized in that the diagnosis is performed by means of immunological techniques. 6. Use of the isolate Nc-Spain 7 according to claim 5, characterized in that the immunological technique is based on the use of antigens from the isolate Nc-Spain 7 in any of its stages of the biological cycle. 7. Use of the isolate Nc-Spain 7 according to claim 6, characterized in that the antigen of the isolate Nc-Spain 7 is formed by the isolate itself in any of its stages of the biological cycle, extracts and/or cell lysates obtained therefrom, antigenic polypeptides obtained from the isolate Nc-Spain 7 or produced by means of recombinant DNA techniques from partially or completely purified polynucleotides obtained from the isolate itself, or mixtures thereof. 8. Use of the isolate Nc-Spain 7 according to claim 7, characterized in that the immunological technique used is an immunoenzymatic technique, an immunofluorescence technique, an immunoblot technique, an agglutination technique or any other method based on the detection of antibodies specific for N. caninum using antigens from the isolate of the invention in any of the stages of the biological cycle. 9. Use of the isolate Nc-Spain 7 according to claim 5, characterized in that the immunological technique is based on the use of antibodies that specifically react against antigens from the isolate Nc-Spain 7 in any of the stages of the biological cycle. 10. Use of the isolate Nc-Spain 7 according to claim 9, characterized in that the immunological technique used is an immunoenzymatic technique, an immunofluorescence technique, an immunoblot technique, an agglutination technique or any other method based on the detection of antigens specific for N. caninum using polyclonal or monoclonal antibodies developed against antigens of the isolate of the invention in any of the stages of the biological cycle. 11. Use of the isolate Nc-Spain 7 according to claims 9 and 10, characterized in that the antibodies are monoclonal. 12. Use of the isolate Nc-Spain 7 according to claim 4, characterized in that the diagnosis is performed by means of nucleic acid detection techniques. 13. Use of the isolate Nc-Spain 7 according to claim 12, characterized in that the nucleic acid detection techniques are based on the specific hybridization and include the application of PCR, in situ hybridization and in situ PCR. 14. Use of the isolate Nc-Spain 7 according to claim 13, characterized in that the nucleic acid detection techniques are applied to the DNA of the parasite extracted from a biological sample. 15. Use of the isolate Nc-Spain 7 of claim 1 in the preparation of a vaccine for the treatment and/or prevention of the infection caused by Neospora in animals. 16. Use of the isolate Nc-Spain 7 according to claim 15, characterized in that the manufacture of the vaccine is performed from the isolate Nc-Spain 7 in any of the stages of the biological cycle. 17. Use of the isolate Nc-Spain 7 according to claim 16, characterized in that the manufacture of the vaccine is performed from the attenuated live, inactivated and/or fixed isolate Nc-Spain 7 in any of the stages of the biological cycle. 18. Use of the isolate Nc-Spain 7 according to claim 16, characterized in that the manufacture of the vaccine is performed from a cell extract and/or lysate of the isolate Nc-Spain 7 in any of the stages of the biological cycle. 19. Use of the isolate Nc-Spain 7 according to claim 16, characterized in that the manufacture of the vaccine is performed by means of the use of antigenic proteins of the isolate Nc-Spain 7 in any of the stages of the biological cycle. 20. Use of the isolate Nc-Spain 7 according to claim 19, characterized in that the antigenic proteins are recombinant proteins. 21. Use of the isolate Nc-Spain 7 according to claims 19 and 20, characterized in that the proteins are partially or completely purified. 22. Use of the isolate Nc-Spain 7 according to claim 15, characterized in that the manufacture of the vaccine is performed by means of the introduction of the gene encoding the immunogen in the host cells of the animals to be vaccinated. 23. Use of the isolate Nc-Spain 7 according to claim 22, characterized in that the gene encoding the immunogen is introduced in the host cell by means of its cloning into a plasmid vector. 24. Use of the isolate Nc-Spain 7 according to claim 22, characterized in that the gene encoding the immunogen is introduced in the host cell by means of its cloning into a virus. 25. Use of the isolate Nc-Spain 7 according to claims 15 to 24, characterized in that the vaccine is prepared as an injectable in liquid formulations or in solid form for its subsequent reconstitution. 26. Use of the isolate Nc-Spain 7 according to claims 15 to 24, characterized in that the vaccine is prepared as an oral formulation. 27. Use of the isolate Nc-Spain 7 according to claims 25 and 26, comprising an adjuvant and/or one or several cytokines and/or immunostimulators. 28. Use of the isolate Nc-Spain 7 of claim 1 in the preparation of a pharmaceutical product for the treatment of the infection and/or disease caused by N. caninum by means of passive immunotherapy, characterized in that it contains antibodies that specifically react against antigens of the isolate Nc-Spain 7. 29. Parasite isolated from bovine nervous tissue, of the species Neospora caninum, described in claim 1, characterized in that is genetically modified. 30. Use of biologically pure cell cultures infected by the genetically modified isolate of claim 29 for obtaining tachyzoites, bradyzoites, polypeptides, antigens, antibodies, nucleic acids and other compounds of use in the preparation of products for the diagnosis, treatment and prevention of neosporosis. 31. Use of the genetically modified isolate of claim 29 in the diagnosis of the infection caused by Neospora. 32. Use of the genetically modified isolate according to claim 31, characterized in that the diagnosis is performed by means of immunological techniques based on the use of antigens from the genetically modified isolate in any of its stages of the biological cycle and/or in the use of antibodies that specifically react with antigens of this genetically modified isolate. 33. Use of the genetically modified isolate of claim 29 in the manufacture of a vaccine for the treatment and/or prevention of the infection caused by Neospora in animals. 34. Use of the genetically modified isolate according to claim 33, characterized in that the manufacture of the vaccine is performed from this genetically modified isolate in any of its stages of the biological cycle as an attenuated live, inactivated and/or fixed genetically modified isolate, or from a cell extract and/or lysate obtained from the genetically modified isolate. 35. Use of the genetically modified isolate of claim 29 in the manufacture of a pharmaceutical product for the treatment of the infection and/or disease caused by N. caninum by means of passive immunotherapy, characterized in that it contains antibodies that specifically react against antigens of the isolate Nc-Spain 7.
