Quadruple inactivated vaccine and preparation method thereof
An inactivated vaccine and inactivated technology, applied in biochemical equipment and methods, vaccines, multivalent vaccines, etc., can solve the problems of poultry adenovirus farming losses and other issues
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Embodiment 1 4
[0060] The antigen composition of embodiment 1 quadruple inactivated vaccine
[0061] The antigens of the quadruple inactivated vaccine are inactivated Newcastle disease La Sota virus strain, inactivated H9 subtype avian influenza TL18 strain, inactivated mutant bursal virus VP2 protein and inactivated group I avian adenovirus type 8b Hexon protein; the nucleotide sequence of the mutant strain bursal virus VP2 protein is as shown in SEQ ID NO.1, and the amino acid is as shown in SEQ ID NO.2; the nucleus of the I group 8b type avian adenovirus Hexon protein The nucleotide sequence is shown in SEQ ID NO.3, and the amino acid is shown in SEQ ID NO.4; the content of the chicken Newcastle disease La Sota virus strain is more than or equal to 10 8.5 EID 50 / 0.1mL; the content of the H9 subtype avian influenza TL18 strain ≥10 7.5 EID 50 / 0.1mL; the agar amplification titer of the VP2 protein content of the mutant strain bursal bursal virus ≥ 1:128; the agar amplification titer of ...
Embodiment 2 4
[0065] The preparation of embodiment 2 quadruple inactivated vaccine
[0066] S1. Isolation and identification of mutant bursal virus
[0067] Collect the bursa tissue of the diseased chicken, grind it, add double-antibody-containing PBS to dilute it into a suspension, add an equal volume of chloroform to the suspension, place it in a low temperature environment at 20 rpm for 24 hours, and take the supernatant after centrifugation at 3000 rpm Packed in ampoule vials for later use. The processed samples were inoculated into 10-day-old chicken embryos with the choriourin pathway (CAM), and incubated at 37°C. The chicken embryos that died within 24 hours were discarded, and the chicken embryos that died after 24 hours were placed at 4°C, the allantoic fluid was collected for hemagglutination test, and the embryo bodies and urea membranes were collected at the same time, which were aseptically ground and then packed in ampoules for later use. The total RNA of the culture was ext...
Embodiment 3
[0103] The safety test of embodiment 3 vaccines
[0104] According to the preparation method of the quadruple inactivated vaccine of Example 1, two batches of quadruple inactivated vaccines are prepared, and the batch numbers are respectively NABFs01 and NABFs02, and safety tests are carried out respectively:
[0105] (1) Single-dose one-time vaccination safety test
[0106] Get 10 days old SPF (no specific pathogen) chicken, the injection dosage of described quadruple inactivated vaccine is 0.3mL / , 10 / group, totally three groups, the first group injection batch number is the quadruple inactivated vaccine of NAIBFs01 , the second group was injected with quadruple inactivated vaccine with batch number NAIBFs02, and the third group was injected with 0.3 mL of normal saline in the neck of 10 animals;
[0107] Get the AA white-feathered broiler chicken (broiler for short) of 7 days of age simultaneously, the injection dosage of described quadruple inactivated vaccine is 0.3mL / , 1...
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