Strain of duck viral hepatitis virus and application thereof
A duck viral hepatitis and virus technology, applied in the direction of antiviral agent, virus/phage, antiviral immunoglobulin, etc., can solve the problem of inability to provide cross-protection ability, and achieve the effect of avoiding infection and improving immunogenicity
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Embodiment 1
[0032] Example 1 Obtaining and Characteristic Determination of Duck Viral Hepatitis Virus
[0033] 1. Virus Isolation
[0034] The liver tissue of ducklings suspected of being infected with duck viral hepatitis was ground and homogenized to obtain disease material samples. Add 1 / 5 times the volume of normal saline of the disease material sample, penicillin and streptomycin with a final concentration of 200 units, freeze and thaw repeatedly 3 times, and centrifuge at 7000 r / min for 20 min to absorb the supernatant. Filter the supernatant with a microporous membrane, take the filtrate and inoculate 10 10-12-day-old duck embryos through the allantoic cavity, discard the dead embryos within 24 hours; collect the dead embryos within 24-96 hours, collect the liver tissue and grind them into After homogenization, mix with allantoic fluid to form a suspension, freeze and thaw repeatedly 3 times, take the supernatant, confirm sterility by sterility test, and use it as a virus sample f...
Embodiment 2
[0053] Embodiment 2 duck viral hepatitis virus vaccine and application thereof
[0054] 1 Virus culture
[0055] The virus seed (Example 1) was diluted 100 times with normal saline, inoculated with 11-13 day-old duck embryos, 0.2 mL per embryo, cultured at 37°C, and discarded duck embryos that died within 24 hours. The incubation time was 96 hours, and the allantoic fluid and embryo body of dead duck embryos were collected. The embryo body was removed from the gallbladder, homogenized, mixed with allantoic fluid, centrifuged at 5000 r / min for 10 min, and the supernatant was collected as the DHV-JS strain virus liquid and stored at -20°C.
[0056] 2 Virus content
[0057] The DHV-JS strain virus solution was diluted 10 times with normal saline, and inoculated into the chorioallantoic cavity of 11-13-day-old duck embryos, 0.2 mL per duck embryo, and used as a blank control. Five duck embryos were inoculated for each dilution, sealed with paraffin, and placed in a 37°C incubat...
Embodiment 3
[0086] Example 3 Preparation and Application of Anti-Duck Viral Hepatitis Egg Yolk Antibody
[0087] The duck viral hepatitis virus vaccine (DHV-JS strain) prepared in Example 2 was used to prepare egg yolk antibodies against duck viral hepatitis.
[0088] 1. Immune laying hens
[0089] Healthy laying hens were used, and the duck viral hepatitis virus vaccine was injected into the chest muscle at multiple points, 0.5ml / hen, which was the first immunization. Ten days after the first immunization, each laying hen was immunized with 1.0ml duck viral hepatitis virus vaccine. 20 days after the first immunization, each laying hen was injected with duck viral hepatitis virus vaccine 2.0ml. After three times of immunization, regularly check the neutralization test titer of egg yolk to duck embryo (the detection method refers to the "2000 edition of Chinese Veterinary Pharmacopoeia"), and the neutralization titer reaches 2 11 , start collecting free eggs.
[0090] 2. Preparation of...
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