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Gene VII type Newcastle disease virus (NDV) marker vaccine strain and application thereof

A vaccine strain and gene technology, applied in the direction of antiviral agents, viral antigen components, viruses/phages, etc., can solve the problems of reducing the virulent carrying amount and infection rate of NDV, and achieve good immune protection effect, wide application value, inhibiting Detoxification effect

Active Publication Date: 2018-01-26
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional vaccines cannot provide ideal immune protection against the current virulent strains of NDV. Domestic and foreign studies have proved that the use of gene-VII vaccines consistent with the epidemic strains can effectively reduce the carrying amount and infection of virulent NDV in immunized chickens. rate, therefore, it is necessary to develop new alternative vaccines to deal with the prevalence of Newcastle disease

Method used

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  • Gene VII type Newcastle disease virus (NDV) marker vaccine strain and application thereof
  • Gene VII type Newcastle disease virus (NDV) marker vaccine strain and application thereof
  • Gene VII type Newcastle disease virus (NDV) marker vaccine strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Rescue of aSG10 strain

[0047] 1. Construction of full-length cDNA of SG10 strain genome

[0048] (1) Purification of the virus In order to obtain a single virus clone, the SG10 strain (disclosed in the literature Liu MM, et al. Generation by reverse genetics of an effective attenuated Newcastle disease virus vaccine based on a prevalent highly virulent Chinese train) .BIOTECHNOL LETT 2015 2015-06-01; 37(6):1287-96.) was purified, the detailed steps are as follows: the virus liquid was diluted 10 times, and after dilution, 100 μL of each titer virus liquid was inoculated with SPF For each dilution, 4 chicken embryos were collected, and the chicken embryos that died within 24 hours were discarded, and the allantoic fluid of the chicken embryos was harvested 4 days later to measure the HA activity. Select the allantoic fluid with the highest dilution factor of HA activity to do the same multiple dilution and inoculation. After the virus was continuously passa...

Embodiment 2

[0085] Example 2 Rescue of aSG10-mHN labeled vaccine strain

[0086] 1. Test method

[0087] 1.1 Construction of full-length cDNA of aSG10-mHN strain genome

[0088]The inserted molecular tag (CACCACCACCAC) is located at fragment D of the full-length cDNA plasmid pOK-aSG10. Design primers (Table 7), insert a molecular tag on the D fragment of pOK-aSG10 (the 5' non-coding region of the HN gene, located at position 8140 in the whole genome) by fusion PCR, and combine the mutated fragment mD with the full-length The D fragment of plasmid pOK-aSG10 was replaced to construct the full-length genome plasmid pOK-aSG10-mHN containing molecular tags. See the construction diagram image 3 .

[0089] Table 7 Primers used to amplify mD fragments

[0090]

[0091] Note: The bold part is the stop codon of HN gene ORF, and the underlined part is the insertion mutation part.

[0092] The fragment after PCR cloning with primers D-U and mHN-L was named mD1, and the fragment after PCR cl...

Embodiment 3

[0109] Example 3 aSG10-mHN labeled vaccine strain to SPF chicken safety test and challenge protection test

[0110] 1 Test method

[0111] 1.1 Safety test of aSG10-mHN labeled vaccine strain on SPF chickens

[0112] In order to measure the safety of the aSG10-mHN labeled vaccine strain rescued by Example 2 of the present invention on SPF chickens, the 2-week-old SPF chickens reared in isolators were randomly divided into 4 groups, 19 in each group. The first group is the aSG10-mHN immunization group, the second group is the aSG10 immunization group, and each chicken is treated with 10 6.0 EID 50 Dosage: Inoculate aSG10-mHN or aSG10 by nasal drop or eye drop respectively. The third group was the blank control group, which was replaced by an equal volume of normal saline during immunization. On the 3rd and 7th days after inoculation, 3 chickens in each group were randomly killed, the gross pathological changes were recorded, and the spleen, thymus, bursa of Fabricius, Harder...

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Abstract

The invention provides a gene VII type NDV marker vaccine strain aSG10-mHN. The marker vaccine strain is an attenuated strain aSG10 in the noncoding region of the 5' terminal of a HN gene and with a nucleotide sequence as shown in SEQ ID No. 1. The marker vaccine strain constructed in the invention has obviously reduced virulence and is genetically stable. Results of immunoprotection tests show that the marker vaccine strain has good immunoprotection effect on NDVs, can effectively inhibit and eliminate viruses and is applicable to prevention of currently epidemic gene VII type NDVs. Through aPCR method, animals infected by wild viruses and vaccinated animals can be discriminated; so the marker vaccine strain is of important significance to monitoring, diagnosis, control and purificationof ND and has good application value.

Description

technical field [0001] The invention belongs to the field of veterinary biological products, in particular to a gene type VII NDV labeled vaccine strain aSG10-mHN and its application. Background technique [0002] Newcastle Disease (ND) is caused by Newcastle Disease Virus (NDV), and is an acute, febrile, highly contagious infectious disease characterized by damage to the digestive tract and respiratory tract of poultry. It spreads fast, has a high mortality rate, and is seriously harmful, causing huge economic losses to the poultry industry of various countries every year. Vaccine immunization is an important means to prevent Newcastle disease, but most of the vaccine strains currently used in China are genotype I or II, which are significantly different from the current prevailing genotype VII strains. Although traditional vaccines can induce a higher level of immune antibodies and resist Newcastle disease virus infection to a certain extent, they cannot completely preven...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/86A61K39/17A61P31/14C12Q1/70C12Q1/6869C12N15/11C12R1/93
Inventor 张国中赵静靳继惠邵梦瑜
Owner CHINA AGRI UNIV
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