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Quadruple vaccine for poultry and preparation method and application thereof

A technology for poultry and vaccines, which is applied in the field of quadruple vaccines for poultry and its preparation, can solve the problems of aggravating stress and disease susceptibility, increasing manpower and material resources, and reducing the production performance of chicken flocks, and achieves good antigenicity and safety. The effect of high sex and long duration of immune protection

Pending Publication Date: 2020-05-15
WUHAN KEQIAN BIOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Most of the current commercialized vaccines are traditional vaccines, and the strains selected were isolated earlier, which are quite different from the current epidemic strains in terms of genes and antigenicity (especially RNA viruses that are highly mutated). Due to the existence of immune deviation, even if chicken farms are repeatedly immunized, it is difficult to control the development of the epidemic. At the same time, it also aggravates the susceptibility to stress and disease, reduces the production performance of chicken flocks, increases the input of manpower and material resources, and further economic losses. increase

Method used

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  • Quadruple vaccine for poultry and preparation method and application thereof
  • Quadruple vaccine for poultry and preparation method and application thereof
  • Quadruple vaccine for poultry and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] This embodiment provides the preparation method of chicken Newcastle disease, avian influenza, infectious bursal disease and avian adenovirus disease quadruple vaccine, figure 1 Be the flowchart of this preparation method, concrete steps comprise:

[0038] (1) Sources of poisonous (bacteria) species used in production

[0039] This product is produced with chicken Newcastle disease virus La Sota strain, H9 subtype avian influenza virus HB01 strain, recombinant Escherichia coli Rosetta (DE3)-rVP2 strain expressing infectious bursal disease virus VP2 protein, expressing avian adenovirus (group I, 4 Type) Fiber2 protein recombinant Escherichia coli Rosetta (DE3)-rFiber2 strains were identified, kept and supplied by Wuhan Keqian Biotechnology Co., Ltd.

[0040] Among them, the preservation number of the H9 subtype avian influenza virus HB01 strain is CCTCC NO: V202017, and it was preserved in the Chinese Type Culture Collection Center of Wuhan University on January 07, 202...

Embodiment 2

[0074] In this embodiment, the quadruple vaccine prepared in Example 1 is tested, and the specific steps are as follows:

[0075] 1 character

[0076] 1.1 Appearance Milky white homogeneous emulsion.

[0077] 1.2 Dosage form It is water-in-oil type. Take a clean straw, suck a small amount of vaccine and drop it in cold water. Except for the first drop that diffuses in the form of cloud, each subsequent drop should not spread.

[0078] 1.3 Stability Take 10ml of the vaccine and put it into a centrifuge tube, centrifuge at 3000r / min for 15min, the water precipitated at the bottom of the tube should not be more than 0.5ml.

[0079] 1.4 Viscosity is measured according to the appendix of the current "Chinese Veterinary Pharmacopoeia", and it should meet the regulations.

[0080] 2 Loading volume inspection: Measure according to the appendix of the current "Chinese Veterinary Pharmacopoeia", and it should meet the regulations.

[0081] 3 Sterility test According to the current "...

Embodiment 3

[0091] 1. Sequencing and sequence analysis of H9 subtype avian influenza virus HB01 strain

[0092] According to the genome sequence of the avian influenza virus in the GeneBank database, primers were designed to amplify the HA and NA gene fragments of the HB01 strain. The primer sequences are as follows:

[0093] HA-F: 5'-GGGAGCAAAAGCAGGGG-3' (SEQ ID NO.1);

[0094] HA-R: 5'-GGAGTAGAAACAAGGGTGTTTT-3' (SEQ ID NO.2);

[0095] NA-F: 5'-GGGAGCAAAAGCAGGAGT-3' (SEQ ID NO.3);

[0096] NA-R: 5'-GGAGTAGAAACAAGGAGTTTTTT-3' (SEQ ID NO. 4).

[0097] After sequencing the HA and NA gene fragments, the genetic evolution information of the HA and NA genes were analyzed by MEGA6 software.

[0098] Comparing the HA gene sequence of the HB01 strain with the nucleotide sequences of 16 representative H9 subtype avian influenza virus reference strains registered on GenBank, it can be seen that the homology between the HB01 strain and the early epidemic strains is low, and it is similar to The ...

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Abstract

The present invention relates to a quadruple vaccine for poultry and a preparation method and an application thereof. The quadruple vaccine for the poultry is for chicken Newcastle disease, H9 subtypeavian influenza, infectious bursal disease and avian adenovirus disease. Inactivated chicken Newcastle disease virus La Sota strain, inactivated H9 subtype avian influenza virus HB01 strain, infectious bursal disease virus capsid protein VP2 and avian adenovirus structural protein Fiber2 are mixed to prepare an oil-water emulsion. The provided quadruple vaccine for the poultry has high safety, good stability and long duration of immune protection, can provide long-lasting maternal antibodies to the offspring young chicken after immunization of hens, and is extremely low in endotoxin content and small in immune side reactions. In addition, the four antigens have good compatibility, competition and expression inhibition between the different antigens do not exist, and the four components inthe quadruple vaccine for the poultry can all reach the same immune effect as respective monovalent inactivated vaccines.

Description

technical field [0001] The invention relates to the field of veterinary biological products, in particular to a quadruple vaccine for poultry and its preparation method and application. Background technique [0002] In the chicken breeding industry, avian influenza (subtype H9) and chicken Newcastle disease have always been at a relatively high incidence rate, especially chicks and laying hens are at a stage of weak resistance and are very prone to infection. Once it occurs, the farm will be given Causing huge economic losses, vaccine prevention is one of the key measures for farms to prevent and control the occurrence of avian influenza (H9 subtype) and Newcastle disease, and it is also the method generally adopted by farms. [0003] At present, the gene of infectious bursal disease virus is constantly mutating, cases of infection with super-virulent strains continue to appear, and there are more and more cases of clinical immune failure. There are large differences in gen...

Claims

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Application Information

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IPC IPC(8): A61K39/17A61K39/145A61K39/235A61K39/12A61P31/14A61P31/16A61P31/20
CPCA61K39/12A61P31/14A61P31/16A61P31/20A61K2039/552A61K2039/5252A61K2039/70C12N2760/18134C12N2760/16134C12N2710/10234C12N2720/10034Y02A50/30
Inventor 徐高原张丽华明凡周明光白红英高尚姚蓉苏秀婵王鑫程泰烺
Owner WUHAN KEQIAN BIOLOGY CO LTD
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