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Seneca valley virus SVV-ZM-201801 and application thereof

A technology of SVV-ZM-201801 and Seneca, which is applied in the fields of biotechnology and food products, can solve problems such as economic losses, lack of vaccines, and troubles in the pig breeding industry, and achieve low pathogenicity, good immunogenicity, and virus The effect of high titer

Active Publication Date: 2019-04-02
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In 2002, Seneca Valley virus was isolated from PER.C6 cell culture for the first time (SVV-001). Subsequently, pig farms in the United States, Canada and other countries reported the occurrence of Seneca Valley disease. After 2014, Brazil Seneca Valley eruptions have occurred in several countries including , the United States, and China, causing severe economic losses
In recent years, Seneca Valley has been prevalent in many provinces in my country, but there is no effective vaccine on the market, making the disease one of the important diseases that plague the pig breeding industry

Method used

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  • Seneca valley virus SVV-ZM-201801 and application thereof
  • Seneca valley virus SVV-ZM-201801 and application thereof
  • Seneca valley virus SVV-ZM-201801 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Isolation of Seneca Valley Virus

[0039] 1. Tissue collection and processing

[0040] Pig lung, liver, tonsil and other tissues were collected from a pig farm, cut into pieces, added 10 times the volume of DMEM medium containing penicillin (400U / ml) and streptomycin (400μg / ml) to grind, 4°C, 10000rpm centrifuged for 10min, The supernatant of the grinding solution was taken and frozen at -80°C for later use.

[0041] 2. Tissue pathogen detection

[0042]Take 200 μl of the above grinding solution, use Beijing Quanshijin Biological Company DNA / RNA Extraction Kit to extract DNA and RNA according to the instructions, and then use Quanshijin Reverse Transcription Kit to reverse transcribe RNA into cDNA. Using the extracted DNA and reverse-transcribed cDNA as templates, respectively use foot-and-mouth disease (FMDV), swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine pseudorabies virus (PRV), and porcine circovirus type ...

Embodiment 2

[0065] Example 2 Pathogenicity Experiment of Seneca Valley Virus SVV-ZM-201801

[0066] Ten 14-day-old piglets negative for Seneca Valley pathogens and antibodies were screened and randomly divided into 2 groups with 5 piglets in each group. The test group was nasal cavity challenge, and each pig was inoculated with 10 8.5 TCID 50 Passaging virus liquid; each pig in the control group was inoculated with an equal volume of sterile PBS solution nasally. The status of the pigs was observed daily, and the rectal temperature was measured and recorded.

[0067] Observed continuously for 10 days, the body temperature of the test group and the control group all fluctuated within the normal range, and the observation of clinical symptoms found that the pigs of the test group and the control group had normal appetite, no blisters occurred, and no pigs died, and all survived ( Figure 4 ). The above data showed that the Seneca Valley virus isolate SVV-ZM-201801 was a low-virulence st...

Embodiment 3

[0068] Application of Example 3 Isolated Strain SVV-ZM-201801 in the Preparation of Seneca Valley Virus Live Vaccine

[0069] 1. Seneca Valley Virus Live Vaccine Preparation

[0070] The SVV-ZM-201801 isolate was inoculated in PK-15 cells or BHK-21 cell monolayer or suspension cells at MOI=0.01 for continuous passage, and the virus liquid was harvested. After repeated freezing and thawing for 3 times, the supernatant was collected by centrifugation, and the virus was determined. Titer. After passing the test of sterility and exogenous virus, add a protective agent containing 10% sucrose and 3% gelatin to the virus solution at a volume ratio of 1:1, mix well, and freeze-dry to prepare a vaccine. The virus content per head is not low at 10 6.0 TCID 50 .

[0071] 2. Vaccine safety test

[0072] A total of 12 healthy piglets around 14 days old and negative for Seneca valley antigen and antibody were selected and randomly divided into 4 groups with 3 piglets in each group. Th...

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Abstract

The invention provides a Seneca valley virus SVV-ZM-201801 and an application thereof. Separation is performed from pig tissues, and sub-culture and plaque purification are performed, so that the Seneca valley virus is obtained. The separated strain can be stably proliferated on sub-culture cells, typical cytopathic effects are formed, and the virus titer is as high as 108.5-1010.5TCID50 / mL. The separated strain does not have obvious pathogenicity on piggies. The Seneca valley virus separated strain has good proliferating properties, and is good in immunogenicity. Vaccines prepared from the separated strain can induce the piggies to generate high-level neutralizing antibodies, and powerful technical support is provided for effective prevention and control of the Seneca valley virus.

Description

technical field [0001] The present invention relates to the fields of biotechnology and food products, in particular to Seneca Valley virus SVV-ZM-201801 and its application. Background technique [0002] Seneca Valley Virus (SVV), also known as type A Senecavirus (Senecavirus A), belongs to the Picornaviridae Seneca Valley Virus genus, is a non-enveloped single-strand normal Stranded RNA virus. The total length of the viral genome is about 7.2kb. From the 5' end to the 3' end, there are 5'UTR, a large open reading frame (ORF), 3'UTR and a poly(A). Pigs infected with SVV can cause blisters, ulcers and even death on the nose and hoof crowns of pigs. The clinical symptoms are very similar to foot-and-mouth disease, swine vesicular disease and vesicular stomatitis. [0003] In 2002, Seneca Valley virus was isolated from PER.C6 cell culture for the first time (SVV-001). Subsequently, pig farms in the United States, Canada and other countries reported the occurrence of Seneca V...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N15/40C07K16/10A61K39/12A61K39/42A61P31/14G01N33/569
CPCA61K39/12A61K2039/505A61P31/14C07K16/10C12N7/00C12N2770/00021C12N2770/00034G01N33/56983G01N2469/10
Inventor 李晓冉马良许磊赵晨璐
Owner CHINA ANIMAL HUSBANDRY IND
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