Newcastle disease virus vaccine strain for gene VII type genetic rescue

A Newcastle disease virus and gene technology, applied in the field of veterinary vaccine strains, can solve the problems of short duration of immunity, residual virulence, and impact on immunogenicity, and achieve a good protective effect

Pending Publication Date: 2022-03-11
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, various problems are often exposed during the use of isolated natural attenuated strains, including a large stress response aft...

Method used

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  • Newcastle disease virus vaccine strain for gene VII type genetic rescue
  • Newcastle disease virus vaccine strain for gene VII type genetic rescue
  • Newcastle disease virus vaccine strain for gene VII type genetic rescue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The construction of embodiment 1 gene VII type Newcastle disease virus attenuated strain

[0026] Through reverse genetics technology, four recombinant viruses with NDV-Ⅶ strain as the parent strain were respectively constructed, in which the HN protein gene underwent different combinations of point mutations, including E347K+Q353R+Y304H+D342N, E347K, E347K+ Q353R and Y304H+D342N. According to the immune effect of the above gene recombinant strains, a recombinant strain of Newcastle disease virus with gene type Ⅵ was constructed.

[0027] The specific steps are described below.

[0028] (1) Construction of Newcastle disease virus cDNA infectious clone

[0029] Primers were designed according to the whole genome sequence (the nucleotide sequence of the genome is SEQ ID NO: 1) of the NDV-Ⅶ strain with the preservation number CCTCC NO: V201968, and the whole genome was divided into 5 segments, which were cloned in one step using Red / ET homologous recombination technolo...

Embodiment 2

[0046] The measurement of embodiment 2 chick embryo mean lethal time (MDT), intracerebral pathogenicity index (ICPI) and vein pathogenicity index (IVPI)

[0047] The determination of the virulence and pathogenicity of Newcastle disease virus can be divided according to the determination results of MDT, ICPI and IVPI. According to the standard of OIE, the above three indices were measured for the unmodified parental strain NDV-Ⅶ, and the modified strains NDV-Ⅶb, NDV-Ⅶc, NDV-Ⅶd and NDV-Ⅶe. The results are shown in Table 3 below.

[0048] Table 3: Determination table of pathogenicity index

[0049]

[0050]

[0051] The results showed that, except for YB17-Ⅵ strain, which was characterized by moderate virulence, the other strains were attenuated strains, indicating that the mutation or replacement of HN gene did not affect the pathogenicity of the strains.

Embodiment 3

[0052] Preparation and safety test of embodiment 3 gene type VII NDV live vaccine

[0053] NDV-Ⅶ strain, NDV-Ⅶb strain, NDV-Ⅶc strain, NDV-Ⅶd strain and NDV-Ⅶe strain were respectively prepared into live vaccines and carried out safety test. Add 5% sucrose skim milk 1:1 to the diluted allantoic fluid, the virus content after quantitative dilution is 10 8.0 EID50 / 0.1mL, mix thoroughly and freeze-dry to prepare live vaccine. The prepared live vaccine was tested for sterility according to the appendix of the 2010 edition of "Chinese Veterinary Pharmacopoeia", and the result met the standard without bacterial contamination.

[0054] The safety test was carried out with 1-day-old SPF chickens. Use 20 chicks for each virus strain, each group of 10, and immunize 10 chicks respectively. 7.0 EID50 and 10 8.0 EID50 dose of virus liquid, 5 PBS control groups, immunized by nasal drops and eye drops, raised under the same conditions, observed continuously for 14 days, and recorded th...

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Abstract

The invention provides a gene VII type Newcastle disease virus vaccine strain, which is a gene VII type Newcastle disease virus attenuated strain NDV-VIIb strain, and is prepared by respectively mutating 340-site amino acid, 342-site amino acid, 347-site amino acid and 353-site amino acid of HN protein of a Newcastle disease virus strain with the preservation number of CCTCC NO: V201968 into histidine, asparagine, lysine and arginine; and then constructing through genetic rescue. The gene VII type Newcastle disease NDV-VIIb strain provided by the invention is used for preparing vaccines. The immune effect of the Newcastle disease virus gene VII type attenuated strain provided by the invention is obviously superior to that of a parent strain, and a good protection effect can be provided for the current popular gene VII type Newcastle disease virus.

Description

technical field [0001] The invention belongs to the technical field of veterinary vaccine strains, in particular to a genetically rescued Newcastle disease virus vaccine strain of gene type VII. [0002] technical background [0003] Newcastle disease is an acute highly contagious infectious disease of poultry caused by Newcastle disease virus, which can cause the disease of chickens, pigeons, geese, wild birds and other animals. Since Newcastle disease was first discovered in Indonesia in 1926, it has spread rapidly and become popular in many countries and regions in the world. It has caused great harm to the poultry industry. It is a Class A disease that must be reported as required by the World Organization for Animal Health. A class of animal diseases. [0004] Liu Hualei et al. found that the prevailing Newcastle disease virus in my country is dominated by genotypes VII and VI through continuous monitoring of the epidemic strains of Newcastle disease. Among them, genoty...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N15/85A61K39/17A61P31/14
CPCC12N7/00C07K14/005C12N15/85A61K39/12A61P31/14C12N2760/18121C12N2760/18122C12N2760/18134A61K2039/552
Inventor 孙化露楚电峰于晓璐李振侯玉超孙鹏范根成杜元钊
Owner YEBIO BIOENG OF QINGDAO
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