Novel goose-star virus strain and application thereof
An astrovirus, a new type of technology, applied in the direction of viruses, antiviral agents, virus antigen components, etc., to achieve good commercial development prospects, effective immune protection, and good safety.
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Embodiment 1
[0012] Example 1 Isolation and Identification of GD 0122 Strain
[0013] 1. Epidemiological investigation Since October 2017, goslings in Guangdong, Sichuan, Hubei and other regions have had an infectious disease with high mortality and obvious symptoms of gout. The disease mainly occurs in goslings aged 5 to 15 days, and all breeds of geese are susceptible. Autopsies were performed on dead goslings. The main pathological changes are as follows: thickened pericardium, attached or coated with white crystals, commonly known as " Villi heart", the liver is covered with urate, the ureter becomes thicker, the metabolism is blocked, the kidney is red and swollen or pale and enlarged, the kidney is mottled, the contents of the end of the rectum are accumulated with white loose feces, the gallbladder and spleen are enlarged, the intestinal wall is thinned, severe In some cases, it can be seen that the joints are red and swollen, and there are urate deposits in the joint cavity. Sever...
Embodiment 2
[0035] Preparation of Virus Seeds of GD 0122 Strain
[0036]Dilute the GD 0122 virus strain 10 times, inoculate healthy goose embryos aged 12 to 13 days, 0.2ml / embryo, and culture them in an incubator with 60% humidity and 37°C for 168 hours, and discard goose embryos that died within 24 hours. The goose embryos that died at ~168 hours were taken out immediately, and stored at 4°C. At 168 hours, all live embryos were taken out, stored at 4°C for 12-24 hours, and the allantoic fluid of goose embryos was collected. According to this method, the goose embryo allantoic fluid harvested above was continued for 5 generations in 12-13 day-old goose embryos, which were respectively marked as D2-D6 generations. The virus content of each passage was determined. Will test for sterility and virus content ≥10 5.0 TCID 50 The virus liquid is mixed, quantitatively divided, and stored in a freeze-dried manner.
Embodiment 3
[0038] Preparation of Antigen of GD 0122 Strain
[0039] 1. Take GD 0122 virus seed for inoculation, dilute it 10 times with sterilized PBS (0.1mol / L, pH value 7.2), inoculate healthy goose embryos aged 12 to 13 days, 0.2ml / embryo, seal the pinhole after inoculation, Place at 36-37°C to continue incubation without turning the embryos.
[0040] 2. Incubation and observation After the goose embryos were inoculated, the embryos were photographed twice a day, and the goose embryos that died 24 hours ago were discarded. Until 168 hours, regardless of death or not, all of them were taken out, the air chamber was upright, and cooled at 2-8°C for 12-24 hours.
[0041] 3. Harvesting Take out the cooled goose embryos, sterilize the air chamber with tincture of iodine, then aseptically peel off the egg shells in the air chamber, remove the egg membrane, cut the chorioallantoic membrane and amniotic membrane (to prevent the yolk from breaking), absorb Embryo fluid. Check each goose emb...
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