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A kind of porcine transmissible gastroenteritis virus egg yolk antibody and preparation method thereof

A yolk antibody, infectious technology, applied in the direction of antibodies, antiviral agents, antiviral immunoglobulins, etc., can solve the problems of mutation and unsatisfactory control effects, and achieve high specificity, good application prospects, and strong virulence. Effect

Active Publication Date: 2018-10-12
陕西诺威利华生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, porcine transmissible gastroenteritis disease is mainly prevented by vaccine immunization, and there are also reports of using egg yolk antibody in a small amount, but both of them have unsatisfactory control effects on the disease, which may have a lot to do with the clinical variation of the epidemic strains of the disease relation

Method used

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  • A kind of porcine transmissible gastroenteritis virus egg yolk antibody and preparation method thereof
  • A kind of porcine transmissible gastroenteritis virus egg yolk antibody and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Preparation of Porcine Transmissible Gastroenteritis Virus TGEV-BD Strain Virus Liquid

[0019] Porcine transmissible gastroenteritis-positive small intestine disease material from a pig farm in Hebei Province was frozen, thawed and filtered, and then inoculated at a ratio of 10% into monolayer-covered ST cells cultured by conventional methods, adsorbed at 37°C for 1 hour, and changed to maintain solution, cultivated in a 37°C incubator for 72 hours. When the operation was carried out blindly to the third generation, CPE phenomena such as cell shrinkage, particle increase, clustering, and shedding occurred ( figure 1 ).

[0020] According to the N gene sequence of transmissible gastroenteritis virus in GenBank as a template, a pair of specific primers were designed with the software Primer5.0: TGEV-P1: 5'-AGACAAACTCGCTATCGCATGG-3'; TGEV-P2: 5'-TCTTGTCACATCACCCTTTACCTGC-3 ', at 94°C for 7min, 94°C for 30s, 56°C for 30s, 72°C for 45s, react for 35 cycles, and ...

Embodiment 2

[0023] Example 2 Preparation of Porcine Transmissible Gastroenteritis Virus Inactivated Vaccine

[0024] Take the above porcine transmissible gastroenteritis virus liquid, add β-propiolactone at a concentration of 0.1%, mix well and place it at 4°C for 18 hours, take it out and place it at 37°C for 2 hours to inactivate, and then follow the steps of 1: Add Freund's complete adjuvant and Freund's incomplete adjuvant at a volume ratio of 1, emulsify with a three-way syringe, and prepare porcine transmissible gastroenteritis virus inactivating agents containing Freund's complete adjuvant and Freund's incomplete adjuvant respectively. Live vaccines should be stored at 4°C for later use.

Embodiment 3

[0025] Example 3 Preparation of Egg Yolk Antibody

[0026] Take 20-week-old egg-laying hens and inject them into breast muscles in points. The vaccine emulsified with Freund's complete adjuvant is used for the first time. The second immunization and the third immunization were carried out respectively, and the immunization doses were 2ml / monkey and 4ml / bird respectively; eggs were collected daily from 1 week after the third immunization, and the specific antibody titer of porcine transmissible gastroenteritis virus in egg yolk was determined. Collect egg yolk fluid with an antibody titer above 1:256.

[0027] After the collected eggs were sterilized with 75% alcohol, use a yolk separator to separate the protein, collect the yolk, add 2 times the volume of PBS buffer (0.01M, pH 7.2) and 1 / 2 volume of chloroform to fully shake, and collect after centrifugation The supernatant was successively precipitated with 50% and 35% ammonium sulfate, and finally dissolved in PBS buffer eq...

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Abstract

The invention relates to a porcine transmissible gastroenteritis virus yolk antibody and a preparation method thereof. A hen immunized with an inactivated porcine transmissible gastroenteritis virus TGEV-BD strain having a preservation number of CGMCC No. 13856 is employed, and the yolk antibody is extracted and purified from egg yolks of the hen. The porcine transmissible gastroenteritis virus yolk antibody prepared by adopting the method has the advantages of safety, high specificity, good treatment effect, low production cost, etc., and can effectively prevent and treat diseases caused by porcine transmissible gastroenteritis virus, and has good application prospect.

Description

technical field [0001] The invention belongs to the fields of veterinary medicine and biotechnology, relates to a porcine transmissible gastroenteritis virus yolk antibody and a preparation method thereof, and belongs to the field of veterinary biological products. Background technique [0002] Transmissible gastroenteritis of swine virus (TGEV) belongs to the Coronaviridae family of Coronaviridae. Like porcine epidemic diarrhea virus and porcine rotavirus, it is one of the important pathogens that cause viral diarrhea in piglets. Occurrence of porcine transmissible gastroenteritis (TGE) disease. TGE is an acute, highly contagious disease characterized by vomiting, severe diarrhea, and dehydration, and is highly lethal to newborn piglets less than 1 week old. TGE has occurred widely all over the world. Pigs of different ages and breeds are susceptible to this disease. The mortality rate of 1-week-old piglets after infection exceeds 90%, which has caused huge economic losses...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10C07K16/02A61K39/42A61P31/14A61P1/04A61P1/00
CPCA61K2039/505C07K16/02C07K16/10
Inventor 潘玉董剑辉陈瑞
Owner 陕西诺威利华生物科技有限公司
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