Anti-swine transmissible gastroenteritis virus and porcine epidemic diarrhea virus egg-yolk antibody and preparation method thereof

A porcine epidemic diarrhea, egg yolk antibody technology, applied in the direction of antiviral agents, antiviral immunoglobulins, antibodies, etc., can solve the problem of poor protection of egg yolk antibodies, and achieve good physical properties, good immunogenicity, and stable virulence. Effect

Active Publication Date: 2015-07-22
兆丰华生物科技(南京)有限公司 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There have been reports on the preparation of egg yolk antibodies against porcine transmissible gastroenteritis virus and porcine epidemic diarrhea virus, but with the variation of clinical epidemic strains, the protection of the original egg yolk antibodies is getting worse

Method used

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  • Anti-swine transmissible gastroenteritis virus and porcine epidemic diarrhea virus egg-yolk antibody and preparation method thereof
  • Anti-swine transmissible gastroenteritis virus and porcine epidemic diarrhea virus egg-yolk antibody and preparation method thereof
  • Anti-swine transmissible gastroenteritis virus and porcine epidemic diarrhea virus egg-yolk antibody and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Preparation of porcine epidemic diarrhea virus ZJ08 strain virus liquid

[0031] Take an appropriate amount of porcine epidemic diarrhea-positive small intestine from a pig farm in Zhejiang, scrape the intestinal mucosa and contents, add PBS in a ratio of 1:5 (weight:volume), freeze and thaw three times, centrifuge to get the supernatant, 0.22μm filter membrane After filtration, trypsin with a final concentration of 20 μg / ml was added to the filtrate and treated at 37°C for 1.5 hours.

[0032] Vero cells were inoculated with a monolayer according to the conventional method, and the virus was inoculated according to the proportion of 10%, adsorbed at 37 °C for 1 hour, supplemented with cell maintenance solution (containing 10 μg / ml trypsin), and cultured in an incubator at 37 °C. When it was passed to the 17th passage, there were obvious and stable CPE changes, the cells were rounded, the granules increased, the clusters were like grape clusters, and the cells ...

Embodiment 2

[0047] Example 2 Preparation of porcine infectious gastroenteritis virus HB08 strain virus liquid

[0048] Take an appropriate amount of porcine infectious gastroenteritis-positive small intestine from a pig farm in Hebei, scrape the intestinal mucosa and contents, add PBS in a ratio of 1:5 (weight:volume), freeze and thaw three times, and centrifuge to get the supernatant, 0.22 μm Filter through a membrane, add trypsin with a final concentration of 20 μg / ml to the filtrate, and treat at 37°C for 1.5 hours.

[0049] The ST cells covered with monolayer were inoculated according to the conventional method, the virus was inoculated according to the proportion of 10%, adsorbed at 37 °C for 1 hour, supplemented with cell maintenance solution (containing 10 μg / ml trypsin), and cultured in a 37 °C incubator. In this way, when the cells were passed blindly to the 8th passage, there was a slight CPE change, and when the 17th passage was passed, there was an obvious and stable CPE chang...

Embodiment 3

[0056] Example 3 Preparation of TGEV-PEDV double inactivated vaccine

[0057] TGEV-PEDV double Freund's complete adjuvant vaccine: Take the above TGEV and PEDV virus solution, mix with equal amount of virus, add 0.3% formaldehyde, place it in a constant temperature incubator at 37°C for 24h for inactivation, shake several times during this period, add etc. The amount of Freund's complete adjuvant was emulsified with a glass syringe to prepare a first-immune inactivated vaccine, which was stored at 4°C for later use.

[0058] TGEV-PEDV double incomplete Freund's adjuvant vaccine: take the above-mentioned TGEV and PEDV virus liquid, mix the same amount of virus, inactivate in the same way, add the same amount of incomplete Freund's adjuvant, emulsify, and prepare the vaccine at 4. Store at ℃ for later use.

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Abstract

The invention discloses an anti-swine transmissible gastroenteritis virus and porcine epidemic diarrhea virus egg-yolk antibody. Swine transmissible gastroenteritis virus and porcine epidemic diarrhea virus combined inactivated vaccine is used as an immunogen to immune laying hens, and an egg-yolk antibody is purified from egg yolk, wherein the swine transmissible gastroenteritis virus is swine transmissible gastroenteritis virus HB08 with the collection number being CGMCC No.7807; and the porcine epidemic diarrhea virus is porcine epidemic diarrhea virus ZJ08 with the collection number being CGMCC No.7806. The prepared egg-yolk antibody has good and safe traits. Artificial infection cure rate of the prepared egg-yolk antibody reaches 100% and is obviously higher than cure rate of an egg-yolk antibody prepared from classical IBDV. Clinical case cure rate of the prepared egg-yolk antibody reaches 93.0%. In clinical preventive tests, incidence of diarrhea can be reduced by 17-24% for tested pigs.

Description

technical field [0001] The invention relates to a preparation method of egg yolk antibody, in particular to a preparation method of egg yolk antibody for treating porcine infectious gastroenteritis virus and porcine epidemic diarrhea virus. Background technique [0002] Porcine transmissible gastroenteritis (TGE) and porcine epidemic diarrhoea (PED) are two important contact viral enteric infectious diseases that cause diarrhea in pigs, both of which are characterized by vomiting and severe watery diarrhea. , dehydration and high lethality to piglets up to 2 weeks of age are characterized. There is currently no effective treatment. There are often reports of porcine transmissible gastroenteritis and porcine epidemic diarrhea infection in clinical practice, which seriously restricts the smooth development of the breeding industry. [0003] The mechanism of action of yolk antibody and maternal antibody in colostrum is similar, both of which are passively immunized to enable ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C07K16/02A61K39/42A61P31/14
Inventor 侯艳红王贵华赵亚荣刘明明满坤于萍萍卢会英
Owner 兆丰华生物科技(南京)有限公司
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