A chimeric recombinant vaccine strain of porcine reproductive and respiratory syndrome and its application

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, and achieve good virulence and genetic stability, wide application value, and pathogenicity. low sex effect

Active Publication Date: 2022-07-12
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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  • A chimeric recombinant vaccine strain of porcine reproductive and respiratory syndrome and its application
  • A chimeric recombinant vaccine strain of porcine reproductive and respiratory syndrome and its application
  • A chimeric recombinant vaccine strain of porcine reproductive and respiratory syndrome and its application

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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 Obtaining the cDNA of the classic PRRSV WH strain

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

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

[0040] Table 1 Reverse transcription system:

[0041] component Volume (μL) 5 x Primescript RT master mix 2 RNA 8

[0042] Reaction program: 37°C for 15min, 85°C f...

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 pigs in each group. They were chimeric virus P10 infection group, chimeric virus P20 infection group, chimeric virus P40 infection group and control group, respectively. The 4 experimental groups were kept in isolation in different animal rooms. The infected group was inoculated with 2ml (1×10 5 TCID 50 / ml) chimeric strain rWH-SD virus solution, the control group was inoculated with 2 ml of 2% DMEM medium, and the clinical symptoms of the experimental piglets were observed every day, and the changes in body temperature and body weight were recorded continuously until the end of the test. At the same time, the blood and nasal swabs of each experimental pig were collected at 0, 3, 5, 7, 10, 14, 21, and 28 days after infection, and the viral nucleic acid in the blood and nasal swab samples was...

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Abstract

The invention relates to the technical field of veterinary biological products, in particular to a chimeric recombinant vaccine strain of porcine reproductive and respiratory syndrome and its application. The porcine reproductive and respiratory syndrome chimeric recombinant vaccine strain is obtained by replacing the GP5 gene, the M protein gene, and the N protein gene of the porcine reproductive and respiratory syndrome WH strain with the genes corresponding to the NADC30-like strain. The chimeric recombinant vaccine strain for porcine reproductive and respiratory syndrome provided by the present invention has low pathogenicity and stable characters, will not cause piglets to become ill after 40 passages, and has high safety; and the immune protection effect is good, and can simultaneously immunize Porcine reproductive and respiratory syndrome and NADC30‑like infection are of great value in the development of novel bivalent vaccines.

Description

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

Claims

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Application Information

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Patent Type & Authority Patents(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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