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A pig csfv-pcv double dna vaccine and preparation method thereof

A technology of CSFV-PCV and DNA vaccine, which is applied in the field of porcine CSFV-PCV double DNA vaccine and its preparation, can solve the problems of mixed cross-infection of CSFV and PCV, complicated preparation process, increased cost, etc., and achieves a simple and convenient immunization method Fast, simple preparation process, and the effect of low-temperature preservation of the structure

Active Publication Date: 2022-07-12
XIDIAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, a serious situation is that the mixed infection of CSFV and PCV has brought new difficulties to the prevention and control of the disease in recent years.
On the one hand, PCV infection can destroy the immune system of pigs, and has a certain degree of inhibition on the humoral immune response of CSFV vaccine, resulting in a low level of immune response induced by CSFV vaccine, resulting in the failure of swine fever vaccine immunity
On the other hand, CSFV vaccines currently on the market are mainly live attenuated vaccines, while PCV vaccines are mainly inactivated whole virus vaccines and recombinant vaccines expressing Cap protein of baculovirus. If live CSFV vaccines and / or PCV vaccines are combined with The mixed use of other vaccines can easily lead to virus mutation, which poses a great safety hazard; in addition, the combination of different whole virus vaccines will interfere with the possible immune response and affect the function of each vaccine; in addition, separate the combined vaccines separately. Packaging, mixed or divided injections before use, not only increases the cost, but also increases the complexity of immunity, and cannot meet the needs of market development
More importantly, the CSFV and PCV-2 monovalent vaccines on the market cannot solve the problem of mixed cross-infection of CSFV and PCV in recent years
A kind of classical swine fever-porcine circular double vaccine reported in the current research is prepared by mixing E2 and Cap protein antigens to form a double vaccine. The protein needs to be expressed, extracted and purified, and the activity of the protein expressed in vitro is low. The immune response of the body cannot be induced well, and the preparation process is complicated and the cost is high

Method used

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  • A pig csfv-pcv double dna vaccine and preparation method thereof
  • A pig csfv-pcv double dna vaccine and preparation method thereof
  • A pig csfv-pcv double dna vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0101] Embodiment 2 Immune effect verification

[0102] (1) Inoculation and immunization of mice and blood collection to separate serum

[0103] Forty-two healthy BALB / c mice were randomly divided into 7 groups with 6 mice in each group.

[0104] The mice were inoculated by subcutaneous immunization. The immunization dose in the same group was the same as the immunization method and site. The total amount of the holding vehicle was unchanged, and the volume was 200 μL / mice. On day 0 after immunization, that is, within 6 hours after primary immunization, blood was collected by tail docking, and serum was separated. Second exemption after 7d. After 14 days, the mice were freed for three times, and the blood was collected from the tail to separate the serum. After 21d, the four-immunization booster was performed, and the blood was collected from the tail at 28d to separate serum.

[0105] The serum separation steps are as follows: after taking the blood sample, the blood is n...

Embodiment 3

[0127] Embodiment 3 Security Detection

[0128] In the steps of inoculation immunization of mice and blood collection to separate serum in Example 2, the body weight of each mouse was measured before immunization, and the body weight was weighed at 0d after immunization, that is, within 6 hours after primary immunization. Second exemption after 7d. After 14 days, three exemptions were carried out, and the body weight was weighed. After 21d, four immune boosters were performed, and the body weight was weighed on 28d. To monitor the effect of the vaccine on the body weight of mice, test the results (see appendix Figure 10 ) showed that the vaccine had no effect on the body weight of the mice and did not affect the growth of the mice.

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Abstract

The present invention discloses a porcine CSFV-PCV duplex DNA vaccine, comprising an expression vector into which a gene E2 encoding CSFV structural protein E2, a gene Erns encoding CSFV structural protein Erns, and a PCV-2 immune protein Cap are inserted The gene Cap and the gene Rep encoding the PCV-1Rep protein. The vaccine can express E2, Erns, Cap and Rep proteins in vivo by immunizing animals by subcutaneous injection, and induce the body to produce higher levels of antibodies, thereby preventing and controlling the infection and prevalence of CSFV and / or PCV‑2 and Mixed cross-infection avoids the occurrence of other diseases caused by mixed infection of CSFV and PCV‑2; and the vaccine helps to purify CSFV and PCV‑2 strains in pig farms. The invention also discloses the preparation method of the vaccine, the preparation process is simple and the use is safe.

Description

technical field [0001] The invention relates to the technical field of vaccines, in particular to a swine CSFV-PCV duplex DNA vaccine against swine fever virus and / or porcine circovirus infection and mixed infection and a preparation method thereof. Background technique [0002] Swine fever virus (Classical swine fever virus, CSFV) can cause acute, febrile, high-contact infectious diseases in pigs, mainly characterized by high temperature and microvascular degeneration, causing systemic hemorrhage, necrosis, and infarction. Existing, causing huge economic losses to the global pig breeding industry. Porcine Circovirus (PCV) can cause various diseases such as porcine multisystem wasting syndrome, porcine respiratory syndrome, porcine skin and nephrotic syndrome. The prevention and control of these two swine viral diseases is the current focus of swine disease prevention and control in the pig breeding industry. [0003] Vaccination is an important means of preventing CSFV an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/187A61K39/12A61P31/14A61P31/20C12N15/62C12N15/85C12N5/10
CPCA61K39/12A61P31/14A61P31/20C07K14/005C12N15/85A61K2039/70A61K2039/552A61K2039/54C12N2770/24334C12N2710/24022C12N2750/10022C12N2750/10034C07K2319/00
Inventor 宁蓬勃郭抗抗王忠良杜付玉张伟洁何骏游鑫刘敏龚春梅张象涵夏玉琼
Owner XIDIAN UNIV
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