A kind of anti-type O foot-and-mouth disease vaccine composition and its preparation and application
A vaccine composition and the technology of the composition, which are applied in the directions of biochemical equipment and methods, medical preparations containing active ingredients, microorganisms, etc., can solve the problem that vaccines contain highly toxic pathogenic substances, pollution, viruses are not completely killed or fully Attenuation and other issues
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Embodiment 1
[0075] Protein expression of foot-and-mouth disease with sequences SEQ ID NO.1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23
[0076] 1. Preparation of foot-and-mouth disease gene fragments used as templates
[0077] The O-type foot-and-mouth disease virus I VP4, VP2, VP3, and VP1 genes shown in the sequences SEQ ID NO.1, 3, 5, and 7, and the O-type foot-and-mouth disease virus II VP4 shown in the sequences SEQ ID NO.9, 11, 13, and 15 respectively , VP2, VP3, VP1 genes, the full-length O-type foot-and-mouth disease virus III VP4, VP2, VP3, VP1 genes shown in the sequences SEQ ID NO.17, 19, 21, 23 were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. The full lengths of the synthesized gene fragments are 255bp, 654bp, 660bp, 639bp, 255bp, 654bp, 660bp, 639bp, 255bp, 654bp, 660bp, 639bp, respectively. The foot-and-mouth disease gene template of the present invention is prepared on the basis of the artificially synthesized foot-and-mouth disease gene fragment.
[0078] 2. Co...
Embodiment 2
[0094] Preparation of vaccine composition against O-type foot-and-mouth disease
[0095] Get the O-type foot-and-mouth disease virus I virus-like particles prepared in Example 1, the O-type foot-and-mouth disease virus II virus-like particles and the O-type foot-and-mouth disease virus III virus-like particles, and slowly add them to the adjuvant, and the process of adding is constantly emulsified at 800 rpm Stir with a machine for 12 minutes, mix evenly, and store at 4°C, which is the vaccine composition against type O foot-and-mouth disease. See Table 3 for specific ratios. The adjuvants suitable for the present invention may be adjuvants known to those skilled in the art. In this example, the adjuvant ISA 206 (Seppic, France) was selected.
[0096] Table 3 anti-O-type foot-and-mouth disease vaccine composition composition ratio
[0097] Vaccine 1 Vaccine 2 Vaccine 3 Vaccine 4 Vaccine 5 Vaccine 6 Vaccine 7 Ⅰ(μg / ml) 100 0 0 100 50 100 15...
Embodiment 3
[0099] Immunogenicity Test of Monovalent Vaccine Against Type O Foot-and-Mouth Disease
[0100] 1. Immunization procedure
[0101] 20 6-month-old healthy cattle with negative antibodies were screened by ELISA kit for foot-and-mouth disease O-type antibody, and were randomly divided into 4 groups with 5 cattle in each group. Groups 1-3 are vaccine 1, vaccine 2, and vaccine 3 immunization groups prepared in Example 2 of the present invention, respectively, and group 4 is a PBS control group. The way of immunization in the immunization group was intramuscular injection of 1 ml in the neck, and the control group was immunized with the same amount of PBS. Blood was collected from each cow before vaccine immunization and every week after immunization until 21 days after immunization.
[0102] 2. Detection of antibody level
[0103] The collected serum was tested for related antibodies by using the FMD O-type antibody ELISA detection kit. The results showed that the antibody leve...
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