The Research About The Virus Preserver Of Hog Cholera Lapinised Virus (C Strain) And Marek'S Disease Turkey Herpes Virus (HVT)

The test applyed 6 virus preserves into PK15 cell lines, chicken embryo fibroblast cells, bovine testicle cells containing virus.Through cells morphology observation, screened a virus preserver called I, and determined its Working Concentration 10%.Through creating PK15 passage lines, Thi veosal and virus preserver relating models each other, it established a good base for improving vaccine production titer .First,we settled PK15 passage lines proliferation curve and PK15 cells passage lines containing virus proliferation curve,found that CSFV did not produce CPE in PK15 cells passage lines and accelerated cells fission proliferation.At about 48 hours in the phase of log phase, PK15 cells passage lines containing virus were 131.5% PK15 cells passage lines without virus.Employed direct immunofluorescence in order to screen models about best cells density, best inoculation dose, best inoculation and harvest way. Through establishing virus proliferation curve about control and experiment(containing virus preserve),we found that it wasnot obviously different to detect virus proliferation by direct immunofluorescence ,however the fluorescence plaque of experiment was lighter and denser than that of control, when in the same titres .Further ,we employed living cells dyeing excluding,found that at 60th hour,living cells number containing virus of experiment were 1.22% that of control,however living cells ratio was with one accord.we also confirmed the above result by employing MTT displays.In vaccine production,virus protein concentration of experiment was 22% higher than that of control.Apllying virus preserver into vaccine production of HCLV bovine testicle cell vaccine.we found virus preserver brought about light lesion when adding 20% and brought about good result when adding 10%,The statistics number of rabbit positive reaction in the experiment was double that in the control .As the result of freezing and thawing again and again showed ,6 times by repeating freezing and thawing in the control was critical point .if it was over 6, the control would not qualify whereas the experiment would qualify exceeding 10 times. Cryopreservation for a long time showed that control would qualify in 3 months whereas experiment would qualify in 5 months.the result of bearing heat showed control would qualify in 8 hours at 37℃, whereas experiment would qualify in 10 hours a! 37℃.The result of the observation by electron microscope and major protective antigen gene E2 and E0 sequencing blast showed that virus preserver did not effect HCLV virus morphology and did not produce gene mutation.The results about immunity antibody levels detecting and innoclating strong CSFV show: virus preserver could stimulate primal neutralized antibdy appearance,and retain strong immunity.