A strain of porcine epidemic diarrhea virus and its culture method and application

A technology for porcine epidemic diarrhea and viruses, applied in the direction of antiviral agents, viruses/bacteriophages, and microbial-based methods, which can solve the problems of limited selection and lack of broad immunogenicity

Active Publication Date: 2016-08-10
WENS FOODSTUFF GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, because the vaccines produced by several domestic manufacturers are all PEDV CV777 strains isolated earlier (referred to as CV777 strains in this invention), they lack extensive immunogenicity, and the selection range in production practice is very limited.

Method used

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  • A strain of porcine epidemic diarrhea virus and its culture method and application
  • A strain of porcine epidemic diarrhea virus and its culture method and application
  • A strain of porcine epidemic diarrhea virus and its culture method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Example 1GDSJ / 2012 Virus strains obtained

[0031] 1. Collection and processing of disease materials

[0032] Seven fresh feces of piglets with suspected viral diarrhea in a pig farm in Guangdong Province were collected, diluted with PBS (phosphate buffer saline) at a weight-to-volume ratio of 1:10 after numbering, shaken and mixed evenly, and repeatedly frozen and thawed 3 times , centrifuged at 8000r / min (rotation per minute) for 10 min (minutes), took the supernatant, sterilized it with a filter with a pore size of 0.22 μm (microns), and stored the resulting treated solution in a -70°C refrigerator for later use.

[0033] 2. RT-PCR (reverse transcription polymerase chain amplification reaction) detection of disease materials

[0034] Take 200 μL (microliter) of each of the above treatment solutions numbered 1-7, and use the AxyPrep Body Fluid Virus DNA / RNA Mini Kit produced by Shanghai Baisai Biotechnology Co., Ltd. to extract RNA. The extracted RNA was rever...

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Abstract

The invention discloses a strain of porcine epidemic diarrhea virus and its cultivation method and application. The strain of porcine epidemic diarrhea virus is named Coronaviridae Coronaviridae Porcine Epidemic Diarrhea Virus GDSJ / 2012 (Porcine epidemic diarrhea virus). It was deposited in the China Center for Type Culture Collection on May 15, 2013, with the preservation number CCTCC NO:V201309. The culture of this strain requires the use of serum-free DMEM medium containing trypsin and magnesium chloride. It can be used to prepare PEDV diagnostic reagents and PEDV vaccines, has good immunogenicity, and can make up for the shortage of existing vaccines.

Description

technical field [0001] The invention relates to the field of microbial viruses, in particular to a new strain of porcine epidemic diarrhea virus, its cultivation method and application. Background technique [0002] Porcine Epidemic Diarrhea Virus (PEDV) is the pathogen of porcine intestinal infectious disease that causes vomiting, diarrhea and dehydration as the main clinical symptoms of pigs (Yin Zhen, 1997; B. E. Straw, 2000), different age and Pigs of different breeds are susceptible to infection, but the hazard to suckling piglets is the most serious, with a mortality rate of up to 100%, causing huge economic losses to the pig industry. It is one of the key diseases in the current prevention and control of pig infectious diseases in my country. [0003] At present, there is no specific drug for treating porcine epidemic diarrhea, and the effect of conventional treatment is not good. Therefore, vaccine prevention is still the main method, and the success of the virus stra...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00G01N33/569A61K39/215A61P31/14C12R1/93
Inventor 田小艳宋延华周庆丰潘永飞
Owner WENS FOODSTUFF GRP CO LTD
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