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Egg yolk antibody for preventing and treating porcine epidemic diarrhea and preparation method of egg yolk antibody

A technology for porcine epidemic diarrhea and egg yolk antibody, which is applied in the field of egg yolk antibody to achieve the effects of good immunogenicity and reduced incidence of diarrhea

Active Publication Date: 2017-09-19
BEIJING DAWEIJIA BIOTECH SHARE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no specific drug to treat porcine epidemic diarrhea, so it is necessary to develop biological products that can effectively treat the disease

Method used

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  • Egg yolk antibody for preventing and treating porcine epidemic diarrhea and preparation method of egg yolk antibody
  • Egg yolk antibody for preventing and treating porcine epidemic diarrhea and preparation method of egg yolk antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Evolutionary analysis of porcine epidemic diarrhea virus PEDV / CH / 2014 S gene

[0024] The porcine epidemic diarrhea virus involved in the present invention, named PEDV / CH / BJ / 2014, was deposited in the General Microbiology Center of the China Committee for the Collection of Microorganisms on December 4, 2014, with a preservation number of CGMCC No.10111. Preservation address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0025] Using DNAStar software to analyze the sequencing results, the results showed that the PEDV / CH / BJ / 2014 strain and the Chinese isolate 2011-BJ-1 constituted a new branch, and the American reported strains MEX / 104 / 2013, USA / Colorado / 2013 and KUIDL-PED-2014-007 has high homology; compared with classic reference strains such as Belgian CV777, Korean strain DR13, etc., it is not on the same evolutionary branch, and has undergone obvious mutations, indicating that the newl...

Embodiment 2

[0026] Example 2 Preparation of Porcine Epidemic Diarrhea Virus Liquid

[0027] The PEDV / CH / BJ / 2014 virus solution was tested for sterility, mycoplasma and exogenous virus with reference to the appendix of "Chinese Veterinary Pharmacopoeia", and the results showed no contamination by bacteria, mold, mycoplasma and exogenous virus.

[0028] The 39th passage culture of PEDV / CH / BJ / 2014 was taken, inserted into a confluent monolayer of Vero cells at 0.5%, absorbed for 1.5 h, discarded the virus liquid, supplemented with maintenance solution containing 5 μg / mL trypsin, and incubated at 37 °C, 5% CO 2 Harvest the virus liquid when 90% of the cells develop lesions under the same conditions, measure the virus price, and the virus content is 3*10^7 TCID 50 / mL, the virus was stored at -80°C for later use.

Embodiment 3

[0029] The preparation of embodiment 3 PEDV inactivated vaccine

[0030] Inactivation of PEDV virus: Take the above virus liquid, add 0.2% β-propiolactone, shake well, put it in a refrigerator at 4°C for 24 hours, and shake it every 2 hours during this period. After the inactivation is completed, put it at 37°C for inactivation Degrade β-propiolactone and store at 4°C for later use. Take an appropriate amount of inactivated virus liquid to inoculate healthy Vero cells for inactivation detection, and observe continuously for 7 days. If the cells have no lesions, it means that the virus inactivation is complete.

[0031] PEDV Freund's complete / incomplete adjuvant vaccine: Take the inactivated virus liquid, add the same amount of Freund's complete adjuvant / Freund's incomplete adjuvant, use a glass syringe to connect the three-way valve for emulsification, and prepare the inactivated Vaccines are stored at 4°C for later use.

[0032] Example 3 Preparation and Purification of PED...

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Abstract

The invention discloses an egg yolk antibody for preventing and treating porcine epidemic diarrhea and a preparation method of the egg yolk antibody. The preparation method comprises the following steps: using an inactivated vaccine of a porcine epidemic diarrhea virus as immunogen for immunizing a laying hen; harvesting egg yolk liquid and purifying from the egg yolk liquid to obtain the egg yolk antibody, wherein a porcine epidemic diarrhea virus strain is PEDV / CH / 2014, with a preservation number of CGMCC (China General Microbiological Culture Collection Center) No. 10111. The egg yolk antibody prepared by the preparation method disclosed by the invention has the advantages of good characters, easiness in storage, high safety and good preventing and treating effects; by an in vitro neutralization test, the neutralizing titer of the egg yolk antibody is measured to reach 1:128; by means of a clinical prevention test, the diarrhea incidence of tested pigs can be reduced by 20 percent, and the cure rate of artificial infection reaches 100 percent.

Description

technical field [0001] The invention relates to a yolk antibody, in particular to a yolk antibody for preventing and treating porcine epidemic diarrhea and a preparation method thereof. Background technique [0002] Porcine Epidemic Diarrhea (PED) is a contagious intestinal infectious disease caused by Porcine Epidemic Diarrhea Virus (PEDV), with vomiting, diarrhea and dehydration as the main clinical symptoms. This disease causes the most serious harm to suckling piglets within 2 weeks of age, and the lethality rate can reach 100%, which seriously affects the healthy development of the pig industry. At present, there is no specific drug to treat porcine epidemic diarrhea, so it is necessary to develop effective biological products to treat the disease. [0003] Newborn piglets obtain maternal antibodies by foraging for colostrum to resist infection of epidemic diarrhea virus. The mechanism of action of egg yolk antibodies is similar to that of maternal antibodies in colost...

Claims

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Application Information

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IPC IPC(8): C07K16/10C07K16/02
CPCC07K16/02C07K16/10
Inventor 张美美张志刚董剑辉金忠辉闫雷张晓杰
Owner BEIJING DAWEIJIA BIOTECH SHARE CO LTD
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