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Seneca valley virus strain and application thereof

A Seneca, virus technology, applied in the direction of viruses, viral peptides, antiviral agents, etc., to achieve the effect of good immunogenicity, immune protection, and good immunogenicity

Active Publication Date: 2019-01-11
HENAN CENT FOR ANIMAL DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the understanding and research on SVV at home and abroad are still shallow. So far, there is no commercialized SVV vaccine available. In order to avoid the economic losses caused by SVV infection to the breeding industry, Seneca Valley virus strains were isolated and Seneca Valley virus was developed. Virus epidemic prevention and detection methods are of great significance

Method used

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  • Seneca valley virus strain and application thereof
  • Seneca valley virus strain and application thereof
  • Seneca valley virus strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Isolation and purification of SVV strains

[0050] 1. Sample processing

[0051] Collect the vesicle fluid or vesicle skin of diseased pigs whose SVV fluorescence quantitative PCR test is positive, prepare a 1:5 suspension with MEM, homogenize with a homogenizer for 5 minutes, centrifuge at 8000r / min for 10 minutes at 4°C, and take The supernatant was filtered with a 0.22 μm filter to obtain a sample treatment solution.

[0052] 2. Virus isolation

[0053]Take the BHK-21 cells that are covered with a single layer and in a good growth state to inoculate the sample treatment solution obtained in step 1. Wash BHK-21 cells twice with serum-free D-Hanks solution before inoculation, add 1 mL of the filtered SVV positive suspension to a T 25 cell culture flask, and store in 5% CO 2 , Cultivate in a cell incubator at 37°C for 1 hour, shake the cell culture bottle once every 20 minutes, so that the virus in the treatment solution can fully contact with the cells and...

Embodiment 2

[0077] The identification of embodiment 2 SVV

[0078] 1. RT-PCR detection method and gene sequencing analysis and identification

[0079] Referring to the RT-PCR identification method of SVV isolated strains in Example 1, the 3D gene of SVV was amplified, and the target fragment was purified and recovered and cloned into the pGEM-T vector for sequencing and sequence analysis. The results showed that the SVV gene could be amplified. 3D target gene.

[0080] According to gene sequencing analysis, the SVV-HeNXX / swine / 2017 strain has a homology of 97.4% with the CH-01-2015 strain that was first isolated in Guangdong, my country in 2015, and the SVV-HeNXX / swine / 2017 strain has 2 American strains In one branch, it has the closest homology with USA / IA46008 / 2015, with a nucleotide identity of 99.4% ( figure 2 ).

[0081] 2. Identification of SVV-HeNXX / swine / 2017 strain by fluorescence quantitative PCR (FQ-PCR)

[0082] On the basis of the RT-PCR detection method of SVV above, ref...

Embodiment 3

[0087] Example 3 Establishment of SVV-HeNXX / swine / 2017 strain to challenge mice

[0088] Six 8-week-old Balb / C female mice, 2 were intraperitoneally injected with SVV cell culture stock solution, 2 lived together, and 2 were reared alone as a negative control; the prepared SVV-HeNXX / swine / 2017 strain was injected intraperitoneally To challenge the virus, the dose of challenge was 200 μL per mouse (containing 10 10 TCID 50), and the other two were housed in the same cage. After the challenge, the mice showed symptoms such as curling up, rough coat, and listlessness, and suspected blister-like protrusions appeared on the nose of the challenged mice, and the cohabitation and negative control mice showed no abnormalities. The heart, liver, spleen, lung, kidney, intestine and nose of the challenged mice were all positive for SVV virus nucleic acid, and no SVV pathogen was detected in the tissues of the cohabitation and negative control mice.

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Abstract

The invention discloses a Seneca valley virus strain and an application thereof, wherein the Seneca valley virus SVV-HeNXX / swine / 2017 is deposited in China Center For Type Culture Collection with theserial number of CCTCCNO: V201767. The Seneca valley virus SVV-HeNXX / swine / 2017 has good immunogenicity. It can induce immune animals to produce higher level of immune protection and can be used for preparing vaccine against Seneca Valley Virus Disease. The vaccine has high safety and no detoxification after immunization, and can induce animals to produce higher level of antibody quickly, so as toachieve high-efficient prevention of Porcine Seneca Valley Virus Disease, and has good popularization and application value.

Description

technical field [0001] The invention relates to the field of microbial biotechnology, in particular to Seneca Valley virus strains and applications thereof. Background technique [0002] Seneca valley virus (Senecavalley virus, SVV), also known as Senecavirus A (SenecavirusA, SVA), belongs to the Picornaviridae Senecavirus genus, and has a relatively close genetic relationship with the Picornaviridae and Cardiovirus genus viruses. close. SVV infection can lead to primary vesicular disease, causing blisters and ulcerated wounds on the nose and hoof crowns of pigs, resulting in lameness and even death, which are indistinguishable from clinical symptoms such as foot-and-mouth disease, swine vesicular disease, and vesicular stomatitis. [0003] In 2002, the first SVV strain (SVV-001 strain) was isolated from cell culture medium pollutants by American researchers, and it was suspected that it might come from pig trypsin or fetal bovine serum. Before 2007, SVV was mainly studied...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C07K14/085C07K16/10C12Q1/70C12Q1/686G01N33/569A61K39/125A61P31/14
CPCA61K39/12A61K2039/552A61P31/14C07K14/005C07K16/1009C12N7/00C12N2770/32021C12N2770/32022C12N2770/32034C12Q1/686C12Q1/701G01N33/56983G01N2333/085C12Q2521/107
Inventor 闫若潜王淑娟马震原班付国赵月龙赵雪丽王翠王东方谢彩华王华俊刘影杨海波项朝荣项黎丽董海岚仲伟平于辉
Owner HENAN CENT FOR ANIMAL DISEASE CONTROL & PREVENTION
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