Porcine epidemic diarrhea virus strain MY01 and application thereof

A porcine epidemic diarrhea, MY01 technology, applied in the direction of viruses, antiviral agents, virus antigen components, etc., can solve the problems of low immune protection and poor immunogenicity, and achieve excellent immune effect, strong immunity, morbidity and death. rate reduction effect

Inactive Publication Date: 2016-04-06
INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problems of poor immunogenicity and low immune protection of the above existing porcine influenza virus vaccines, the invention provides a porcine epidemic diarrhea virus MY01 strain with strong immunogenicity

Method used

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  • Porcine epidemic diarrhea virus strain MY01 and application thereof
  • Porcine epidemic diarrhea virus strain MY01 and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The medium for cell culture is DMEM with 10% newborn bovine serum; its preparation: take 500ml of liquid DMEM medium, add newborn bovine serum according to the volume of 10%, mix well and set aside.

[0023] Medium for virus culture, namely, DMEM with 2% newborn bovine serum; its preparation: take 500ml of liquid DMEM medium, add newborn bovine serum according to the volume of 2%, mix well and set aside.

[0024] 1.1 Isolation, purification and passaging of PEDV

[0025] From the diseased materials sent by a large pig farm in Linyi, Shandong Province, collect the intestinal contents of pigs whose test results are positive for PEDV and negative for TGEV and other viruses, centrifuge at 12000r / min for 10min, take the supernatant, and use a 0.22μm filter membrane filter. Filter the supernatant of the disease material according to 1ml / 25cm 2 Inoculate porcine intestinal epithelial cells at the ratio of 2 hours after infection, add 10ml virus culture medium, place at 37°C,...

Embodiment 2

[0112] Application of Porcine Epidemic Diarrhea Virus MY01 Strain in the Preparation of Porcine Epidemic Diarrhea Virus Inactivated Vaccine: Take the Porcine Epidemic Diarrhea Virus MY01 Strain with the preservation number of CGMCC No.11495, inoculate it in PIEC cells to propagate the virus, and collect the virus The virus liquid is inactivated by formaldehyde and mixed with aluminum hydroxide adjuvant to form an inactivated aluminum gel vaccine. The operation steps are as follows:

[0113] 2.1 Selection of poisonous species

[0114] The virus seed for making seedlings is PEDVMY01 strain, and its storage number is CGMCCNo.11495, which has strong pathogenicity to pigs. 0.1mL virus liquid (10 4.0 TCID 50 ) can cause 90% of piglets to be ill by oral inoculation. The immunogenicity of PEDVMY01 strain is good, and the minimum immune dose should be ≥2.0×10 6 TCID 50 .

[0115] 2.2 Preparation of poisonous seeds for production

[0116] 2.2.1 Primary Production Seed Propagation ...

Embodiment 3

[0141] Application of Porcine Epidemic Diarrhea Virus MY01 Strain in the Preparation of Porcine Epidemic Diarrhea Virus Attenuated Vaccine: Take Porcine Epidemic Diarrhea Virus MY01 strain with the preservation number of CGMCC No. 11495, and transfer it to monkey Source VERO cell culture. Plaque purification was performed from passage 20 (48 passages in total), and plaque purification was performed 3 times in a row. Porcine epidemic diarrhea virus attenuated vaccine was prepared with 22 generations of cloned virus (70 generations in total).

[0142] 2.1 Selection of poisonous species

[0143] The seedling virus was PEDVMY01 strain. After subcultured by PIEC nuclei Vero cells, the F70 generation of MY01 strain lost its pathogenicity to piglets, but retained good immunogenicity. The minimum immune dose should be ≥2.0×10 5 TCID 50 .

[0144] 2.2 Preparation of poisonous seeds for production

[0145] 2.2.1 Primary Production Seed Propagation

[0146] Take the PEDVMY01 strai...

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Abstract

The invention relates to a porcine epidemic diarrhea virus strain MY01 and application thereof and belongs to the field of porcine epidemic diarrhea virus vaccine reagents. The porcine epidemic diarrhea virus strain MY01 is collected in China General Microbiological Culture Collection Center under CGMCC No. 11495. The porcine epidemic diarrhea virus strain MY01 is higher in pathogenicity for pigs and good in immunogenicity, a deactivated oil emulsion vaccine prepared using the strain is safe and reliable, homologous attacking protection can be provided, protection is also provided for PEDV (porcine epidemic diarrhea virus) epidemic strains, higher immunity can be generated after immunization, inoculated pigs are significantly lower in morbidity and mortality, the immunization effect is superior than that of existing commercial vaccines in the market, and the strain has the advantage of competing with like products home and abroad, can effectively prevent the prevalence and spread of porcine epidemic diarrhea virus and reduce economic cost caused by the disease and has a promising application prospect.

Description

technical field [0001] The invention relates to a strain of porcine epidemic diarrhea virus MY01 and an application thereof, belonging to the field of porcine epidemic diarrhea virus vaccine reagents. Background technique [0002] Porcine epidemic diarrhea (Porcine epidemic diarrhea, PED) is a highly contagious intestinal infectious disease of pigs caused by porcine epidemic diarrhea virus (PEDV), characterized by vomiting, diarrhea and loss of appetite. susceptible. Porcine epidemic diarrhea virus (PEDV) is a member of the Coronaviridae family. The shape of the virus is slightly spherical, with a capsule, and there are petal-shaped fibrils on the capsule, which is crown-shaped. PEDV nucleic acid is a linear single-stranded positive-strand RNA with a length of 27,000 to 33,000 nucleotides (nt). The virus cannot agglutinate red blood cells from humans, rabbits, pigs, mice, dogs, horses, sheep, and cattle. Weak resistance to the outside world, sensitive to ether and chloro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/215A61P31/14C12R1/93
CPCA61K39/12A61K2039/5252A61K2039/5254A61K2039/55505C12N7/00C12N2770/20021C12N2770/20034
Inventor 吴家强陈智刘少宁杜以军郭立辉陈蕾孙文博丛晓燕任素芳张玉玉于江李加飞王淞
Owner INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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