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Virus recombinant vaccine A-NDV-LX/14 for newcastle disease and construction method thereof

A technology of A-NDV-LX and Newcastle disease virus, applied in the direction of virus/bacteriophage, antiviral agents, virus antigen components, etc., can solve the problem of genotype inconsistency between strains and epidemic strains, and achieve the effect of broad application prospects

Active Publication Date: 2012-07-04
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The first purpose of the present invention is to invent a recombinant Newcastle disease attenuated vaccine expressing gene VII type F and HN, so as to solve the problem of inconsistency between the currently used vaccine strain and the epidemic strain genotype

Method used

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  • Virus recombinant vaccine A-NDV-LX/14 for newcastle disease and construction method thereof
  • Virus recombinant vaccine A-NDV-LX/14 for newcastle disease and construction method thereof
  • Virus recombinant vaccine A-NDV-LX/14 for newcastle disease and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1. Construction of full-length clone prLX of NDV / LX strain.

[0025] build mode like figure 1 shown.

[0026] Step 1: Obtaining NDV / LX strain

[0027] A batch of attenuated Newcastle disease virus strains were isolated from the intestinal tract of healthy poultry, and their biological characteristics: hemagglutination value, ICPI, EID50 were analyzed, and among these attenuated strains, vaccine candidates with high hemorrhagic agglutination value and strong replication ability were screened and carry out immunogenicity analysis to the candidate strains, immunize 2-week-old SPF chickens, measure anti-Newcastle disease virus titers in the serum after immunization and the immune level of the digestive tract and respiratory tract mucosa, and finally obtain good in the respiratory tract and digestive tract Replication, induction of high-titer specific antibodies and mucosal immune response to Newcastle disease attenuated strain NDV / LX (genotype I Newcastle disease...

Embodiment 2

[0047] Construction Step 1: Construction of recombinant plasmid pA-NDV-LX / I4

[0048] 1. Construct a plasmid containing the F gene and the HN gene of the isolate NDV / LX. The primer sequences for amplification are as follows:

[0049] QDr2F: 5'-GCTCGAGACTCGGTATTCATCACCACCTAT-3' (SEQ ID NO.19) Xho I

[0050] LXrg2R: 5'-CGTCTCGGTCTACCTCCACATCAATAGTGAC-3' (SEQ ID NO.20) BsmB I

[0051] QDr3F: 5'-TACTTGAAGACGTAGACCCCGAAGA-3' (SEQ ID NO.21) Bbs I

[0052] QDr3R: 5'-GTCGACACAGATCATTATTTAGTGTCATGG-3' (SEQ ID NO.22) Sal I

[0053] LX3F2: 5'-GTCGACIACTTATAGTTAGTTCGCCTG-3' (SEQ ID NO.23) Sal I

[0054] 5'-CGTCTCGTCGAGATATCAAGATTGCCTGTCAC A-3' (SEQ ID

[0055] LXrg3R: BsmB I

[0056] NO.24)

[0057] LXrg4F5-ACGTCGACTGAGCTTGGGAATGTCAACAACTC-3 (SEQ ID NO.25) Sal I

[0058] LXrg4R 5-CAGTTGACTCAACCGGCTAGACCTGGCTTC-3 (SEQ ID NO. 26) Spe I

[0059] Using the clone prLX containing the full-length genome of the NDV / LX strain as a template, use primers QDr2F and LXrg2R to amplify the fragm...

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Abstract

The invention relates to a virus vaccine A-NDV-LX / 14 for a newcastle disease, which is a recombinant of expressed genes VII F and HN, and the preservation number is CGMCCNO: 3844. After two weeks of the constructed virus A-NDV-LX / 14 immunity test on chicken, the average potency of the serum HI is 27. After four days of eliminating toxic material with immune group LaSota, the virus isolation ratesof SPF chicken laryngotracheal and cloacal swab samples are 10 percent and 30 percent respectively, and the virus isolation rates of chicken laryngotracheal and cloacal swab samples are 40 percent and 20 percent respectively. After four days of eliminating toxic material with immune group V4, the virus isolation rates of SPF chicken laryngotracheal and cloacal swab samples are 30 percent and 10 percent respectively, and the virus isolation rates of chicken laryngotracheal and cloacal swab samples are 30 percent and 20 percent respectively. Compared with traditional vaccine, more effective immune protection can be provided by the recombinant virus vaccine.

Description

technical field [0001] The invention relates to the application of reverse genetic technology and immunological technology to produce a recombinant Newcastle disease attenuated vaccine A-NDV-LX / I4 expressing gene VII type F and HN, which is used for making vaccines. Background technique [0002] Reverse genetics manipulation technique (Reverse genetics manipulation technique) refers to the construction of infectious molecular clones of RNA viruses in vitro, the reverse transcription of viral genomic RNA into cDNA, and various in vitro artificial manipulations on the DNA molecular level. A research technique for assembling new RNA viruses from genomic cDNA and various auxiliary proteins [Neumann G, Whitt M A, Kawaoka Y. Adecade after the generation of a negative-sense RNA virus from cloned cDNA-what have we learned? Journal of General Virology, 2002, 83(11): 2635-2662.], also known as full-length infectious cDNA cloning technology, is often referred to as "virus rescue". Sin...

Claims

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Application Information

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IPC IPC(8): C12N7/01A61K39/17A61P31/14C12R1/93
Inventor 刘秀梵王晓泉刘晓文胡顺林刘文博刘慧谋
Owner YANGZHOU UNIV
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