Method for preparing duck virus hepatitis divalent refined egg yolk antibody

A technology for duck viral hepatitis and viral hepatitis, which is applied in the field of veterinary biological products to achieve the effect of high potency and high protection rate

Active Publication Date: 2012-12-19
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But because the yolk contains a lot of lipids, it brings a lot of inconvenience to use, so people take various methods to extract and isolate IgY from the yolk

Method used

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  • Method for preparing duck virus hepatitis divalent refined egg yolk antibody
  • Method for preparing duck virus hepatitis divalent refined egg yolk antibody
  • Method for preparing duck virus hepatitis divalent refined egg yolk antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] - Vaccine (immunogen) preparation:

[0082] 1. Preparation of virus liquid for seedling production

[0083] (1) YBH1 strain venom preparation Dilute YBH1 strain 100 times, and inoculate 100 10-day-old SPF chicken embryos in the allantoic cavity, 0.2ml per embryo, incubate at 36-37°C, and inspect the embryos twice a day . Chicken embryos that died within 48 to 120 hours after inoculation were placed at 2 to 8°C for 4 to 12 hours, and allantoic fluid, amniotic fluid and embryo bodies were collected, and the heads and limbs were removed from the embryo bodies. Homogenize with embryo fluid, freeze and thaw three times, centrifuge at 4000r / min for 30min, take the supernatant and mix it in a sterile container, store at 2-8°C. (See Table 1).

[0084] (2) Preparation of YBHX strain venom Dilute YBHX strain 100-fold, inoculate 100 susceptible duck embryos 12-13 days old in the allantoic cavity, 0.2ml per embryo, incubate at 36-37°C, irradiate twice a day Embryo inspection. ...

Embodiment 2

[0118] ——Preparation of egg yolk antibody against duck viral hepatitis

[0119] 1. Preparation of High Free Eggs

[0120] The duck viral hepatitis bivalent inactivated vaccine prepared above is immunized laying hen, first immunization every chicken neck subcutaneously injects 1.0ml bivalent inactivated vaccine, carries out the second immunization after 14 days, every chicken neck subcutaneous injection 2.0ml ml of bivalent inactivated vaccine, the third immunization is carried out 14 days after the second immunization, and 2.0ml of bivalent inactivated vaccine is injected subcutaneously in the neck of each chicken, and the third immunization is carried out 14 days after the third immunization, and the neck of each chicken is subcutaneously injected After injecting 2.0ml of bivalent inactivated vaccine, 14 days after the fourth immunization, the egg yolk was collected to determine the titers of duck viral hepatitis 1 and new neutralizing antibody, which were 1:1024 and 1:2048, ...

Embodiment 3

[0135] ——Product inspection of egg yolk antibody

[0136] (1) Properties This product is light yellow transparent liquid. The pH value is 7.0.

[0137] (2) The filling volume inspection is carried out according to the appendix of the current "Chinese Veterinary Pharmacopoeia", which is in compliance with the regulations.

[0138] (3) The sterility test is carried out according to the appendix of the current "Chinese Veterinary Pharmacopoeia", and the growth is sterile.

[0139](4) For safety inspection, 10 susceptible ducklings (type 1 and new duck hepatitis maternal antibodies <1:4) aged 1 to 3 days were used, and 2.0ml of this product was injected into each muscle point. On the 14th day of observation, all susceptible ducklings were healthy and alive.

[0140] (5) Effectiveness test (see Table 6)

[0141] 1) Serological methods Determine the neutralizing antibody titer according to the method in Note 1. The yolk antibody titers of duck viral hepatitis YBH1 strain and YBH...

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Abstract

The invention relates to a method for preparing a duck virus hepatitis divalent refined egg yolk antibody. The method is that vaccines of a duck virus hepatitis virus 1 type YBH1 strain and novel YBHX strain with good immunogenicity are selected from duck virus hepatitis superior prevalent strains to prepare strains, the strains are respectively inoculated to a chicken embryo and a duck embryo, a dead embryo body and the embryo juice are respectively obtained, the virus liquid is collected after grinding and freeze thawing, ultrafiltration concentration and inactivation with formaldehyde solution are carried out, and oil adjuvant is added to mix and emulsify to produce the vaccine. The vaccine is inoculated to a laying hen, the egg with high immunity is obtained, the yolk is separated, inactivated, extracted and refined to produce the duck virus hepatitis divalent refined egg yolk antibody which can prevent the duck virus hepatitis which is caused by the 1 type and novel duck hepatitis viruses, so the egg yolk has the advantages of high efficiency, good safety, high protection ratio and the like.

Description

technical field [0001] The invention relates to a preparation method for preventing type 1 and new type duck viral hepatitis by divalent refined egg yolk antibody, which belongs to the field of veterinary biological products. Background technique [0002] Duck viral hepatitis (DVH) is an acute, highly fatal, contagious disease of ducklings caused by duck hepatitis virus (DHV) with liver as the main pathological change. Clinical manifestations include neurological symptoms such as spasms, convulsions, and opisthotonus, and liver swelling and hemorrhage are characteristic lesions. Ducklings aged 1 to 3 weeks are susceptible to severe disease and spread rapidly. Ducklings usually die within 1 hour after symptoms appear, and the mortality rate of 10-day-old ducklings after infection is 90-95%. It has become a serious hazard to the duck industry. one of the main diseases. [0003] Duck hepatitis virus (DHV) serotype DHV has 4 independent serotypes, type 1, type 2, type 3 and ne...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C07K16/02
Inventor 徐保娟宫晓郭伟伟王龙孙健胡潇
Owner YEBIO BIOENG OF QINGDAO
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