Study Of Recombinant Canine Interferon Alpha Activity Deteciton And Treatment Of Canine Parvovirus Disease

Interferon (IFN) is a kind of excellent antiviral, antitumor and immunomodulatory cytokines,since it has been found, interferon has been widely used in the treatment of human diseases and healthcare. Recombinant canine interferon alpha is containing the recombinant canine interferon alpha generecombinant plasmid was transfected into E.coli, and the high density fermentation, has a better curativeeffect on canine viral disease. In the study of recombinant canine interferon alpha new veterinary drugs,biological activity is the key index for evaluating the effect of good products, only the good activity ofproduct has practical application value in clinical. This paper mainly studies the method for detection ofbiological activity of recombinant canine interferon alpha, and the artificial reproduction of canineparvovirus with different doses of recombinant canine interferon alpha for pharmacodynamic studies,specific studies are as follows:1．During the detecting methods of biological activity of recombinant canine interferon alpha study,MDCK cells were cultured by routine passage method, through the cell concentration optimizationselection test, concentration of each ml containing30~400,000cells, incubator at5%CO2,37℃staticculture for24hours, the recombinant canine interferon alpha single cell culture inoculated with differentconcentrations in the good, incubator at5%CO2,37℃cultured with the VSV virus attack after24hours, and then in37℃and5%CO2incubator culture, end the poison attack when the virus controlhole is more than75%cell lesions. They were stained with crystal violet staining solution,determination the absorbance at the wavelength of570nm, through the four parameter regression dataprocessing method to calculate the sample activity. Research results show that, the test method fordetermining the crystal violet staining was determined at the wavelength of570nm absorbance value,compared with the conventional cell disease method, it is more simple and less operative error. Theprecision andrepeatability experiments show the method has high precision and good reproducibility.2．In order to verify the effect of recombinant canine interferon alpha on viral diseases, we willvaccinate55~70days old beagle dogs with canine parvovirus, which are unvaccinated canineparvovirus vaccine. The obvious clinical symptoms with recombinant canine interferon alpha after thecytopathic effect assay with high biological activity for treatment. Research results show that, with theinfusion, antibacterial, hemostatic, vomiting, fever and other auxiliary measures, recombinant canineinterferon alpha disease effective rate is up to100%according to400,000units per kg of body weighton artificial reproduction of canine parvovirus. Indicated through the treatment of artificial reproductionof canine parvovirus disease results with different doses of recombinant canine interferon alpha,recombinant canine interferon alpha treatment results and dosage were positively correlated, with therecombinant canine interferon alpha dose (expressed by the cell titer) increased, mortality rate increased.Through the three batches of recombinant canine interferon alpha repeat treatment test of artificialreproduction of canine parvovirus disease shows recombinant canine interferon alpha treatment effect isstable for canine parvovirus disease.Through the research, it is determined at the wavelength of570nm absorbance value by cell lesionscombined with crystal violet staining inhibition and dealed with by four parameters regression data processing method, this determination method of biological activity of recombinant canine interferonalpha is of high precision and good repeatability. According to400000units per dose of kg body weight,and with rehydration, antibacterial, hemostatic, vomiting, fever and other auxiliary measures, theartificial reproduction of canine parvovirus disease treatment effect is indeed and stable curative effect.