Chimeric recombinant vaccine strain for porcine reproductive and respiratory syndrome and application thereof

A respiratory syndrome and recombinant vaccine technology, applied in the field of chimeric recombinant vaccine strains for porcine reproductive and respiratory syndrome, can solve problems such as inability to effectively resist infection, achieve good virulence and genetic stability, low pathogenicity, and economical effect of time

Active Publication Date: 2021-07-23
WUHAN KEQIAN BIOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

During clinical application, it was found that the use of PRRSV attenuated live vaccines could not effectively resist the infection of NADC30-like strains. For example, pig farms immunized with VR2332 strain commercial vaccines also broke out HENAN-XINX strains, HENAN-HEB strains, and JL580 strains. Strain infection

Method used

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  • Chimeric recombinant vaccine strain for porcine reproductive and respiratory syndrome and application thereof
  • Chimeric recombinant vaccine strain for porcine reproductive and respiratory syndrome and application thereof
  • Chimeric recombinant vaccine strain for porcine reproductive and respiratory syndrome and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1. Construction of the full-length cDNA cloning plasmid of the classic PRRSV WH strain

[0037] 1.1 Acquisition of cDNA of classic PRRSV WH strain

[0038] The whole genome of the classic PRRSV WH strain was extracted and reverse transcribed into cDNA.

[0039] PRRSV genomic RNA extraction method: Take 300 μL virus liquid supernatant, add 1 mL trizol, blow and mix with a gun, put on ice for 10 minutes; add 200 μL chloroform, shake and mix, put on ice for 3 minutes, 13000rpm 4°C for 15 minutes, take the upper aqueous phase To a new EP tube; add 500 μL of isopropanol, mix gently, put on ice for 30 min; 13000 rpm at 4 °C for 10 min, discard the supernatant; add 1 mL of 75% ethanol, wash the precipitate, 13000 rpm at 4 °C for 10 min, discard the supernatant; air dry , add 30 μL DEPC water.

[0040] Table 1 Reverse transcription system:

[0041] components Volume (μL) 5×Primescript RT master mix 2 RNA 8

[0042] Reaction program: 37°C for 15min,...

Embodiment 2

[0147] 1. Pathogenicity test of chimeric virus rWH-SD P5, P20, P40 infected piglets

[0148] Twenty 28-day-old PRRSV antibody-negative piglets were randomly divided into 4 groups with 5 piglets in each group. They were chimeric virus P10 infection group, chimeric virus P20 infection group, chimeric virus P40 infection group and control group. The 4 experimental groups were kept in isolation in different animal rooms. The infection group was inoculated with 2ml (1×10 5 TCID 50 / ml) chimeric strain rWH-SD virus liquid, the control group was inoculated with 2ml2% DMEM medium, observed the clinical symptoms of the experimental piglets, recorded body temperature and body weight changes every day, and observed continuously until the end of the test. At the same time, the blood and nasal swabs of each test pig were collected at 0, 3, 5, 7, 10, 14, 21, and 28 days after infection, and the virus nucleic acid in the blood and nasal swab samples was detected by RT-PCR method, and the ...

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Abstract

The invention relates to the technical field of veterinary biological products, in particular to a chimeric recombinant vaccine strain for porcine reproductive and respiratory syndrome and application thereof. The chimeric recombinant vaccine strain for the porcine reproductive and respiratory syndrome is obtained by replacing a GP5 gene, an M protein gene and an N protein gene of a porcine reproductive and respiratory syndrome WH strain with corresponding genes of an NADC30-like strain. The chimeric recombinant vaccine strain for the porcine reproductive and respiratory syndrome provided by the invention is low in pathogenicity and stable in characters, does not cause diseases of piglets after 40 times of passage, and is relatively high in safety; and moreover, the chimeric recombinant vaccine strain has a good immune protection effect, can simultaneously immunize against the porcine reproductive and respiratory syndrome and NADC30-like infection, and has important value in the development of new bivalent vaccines.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a porcine reproductive and respiratory syndrome chimeric recombinant vaccine strain and application thereof. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS) is an important viral disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which seriously affects the pig industry. The disease is widespread and causes huge economic losses to the swine industry worldwide. For example, the previous outbreak of highly pathogenic PRRS (HP-PRRS) had high clinical morbidity and mortality, which brought unprecedented economic losses to the existing pig industry. Compared with traditional classic PRRSV, the pathogen is characterized by the presence of There are 30 consecutive amino acid deletions in the Nsp2 gene. Later, a mutant strain of PRRSV appeared. The virus genome was characterized by 131 discontinuous a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/40A61K39/12A61P31/14C12R1/93
CPCC12N7/00A61K39/12A61P31/14C12N2770/10021C12N2770/10022C12N2770/10034
Inventor 徐高原周明光张华伟曾小燕陈映瑾郝根喜周飞燕邵伟金建云
Owner WUHAN KEQIAN BIOLOGY CO LTD
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