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Egg yolk antibody of inclusion body hepatitis and preparation method thereof

A technology of egg yolk antibody and inclusion body, which is applied in the field of egg yolk antibody for chicken inclusion body hepatitis and its preparation, can solve the problem of no effective antibody for chicken inclusion body hepatitis, and achieve a good development effect

Active Publication Date: 2016-11-02
CHONGQING SANJIE ZHONGXIN BIOLOGICAL ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the problem that the existing chicken inclusion body hepatitis has no effective antibody, the present invention proposes an egg yolk antibody for chicken inclusion body hepatitis, which can effectively prevent and treat chicken inclusion body hepatitis, and the cure rate reaches more than 82%.

Method used

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  • Egg yolk antibody of inclusion body hepatitis and preparation method thereof
  • Egg yolk antibody of inclusion body hepatitis and preparation method thereof
  • Egg yolk antibody of inclusion body hepatitis and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Isolation and identification of virus strains

[0025] 1.1 Virus isolation

[0026] In March 2014, livers of dead chickens clinically diagnosed as chicken inclusion body hepatitis were collected from a broiler farm in Dagao Village, Xinzhai Town, Zhangqiu City, Jinan City, Shandong Province, and homogenized with normal saline at a mass-volume ratio of 1:5 (W / V) , repeated freezing and thawing twice at -20°C and 4°C, centrifuged at 12000 rpm for 30 minutes at 4°C, collected the supernatant, sterilized by filtration, aliquoted, and stored at -40°C. Samples were taken at the same time, and the sterility test was carried out according to the current method of "The Veterinary Pharmacopoeia of the People's Republic of China".

[0027] 1.2 PCR identification

[0028] 1.2.1 Primer design

[0029] According to the chicken inclusion body hepatitis group I gene sequence on Genbank, design a pair of primers, the target fragment is about 1300bp, the primer sequence is as follows:...

Embodiment 2

[0046] Virus subculture and identification

[0047] 2.1 Subculture

[0048] The supernatant of chicken inclusion body hepatitis virus SDJ15-8 strain disease material was diluted 10 times with normal saline, inoculated with 7-day-old SPF chicken embryos through the yolk sac, 0.1ml / embryo, and collected the dead chicken embryo allantoic fluid 24-120 hours after inoculation, Marked as chicken inclusion body hepatitis virus SDJ15-8 strain E1, after passing the sterility test, it was subcultured continuously. During the subculture of the virus, the dilution factor of the virus was appropriately adjusted according to the time period of chicken embryo death after each generation of virus inoculation, until the virus was subcultured. After inoculation, more than 60% of chicken embryos died in 60-92 hours. When the virus was subcultured to the 7th generation, more than 70% of the chicken embryos died in 60-92 hours, and the allantoic fluid was collected, labeled as chicken inclusion b...

Embodiment 3

[0079] Preparation of Refined Egg Yolk Antibody of Chicken Inclusion Body Hepatitis Virus Serum Type 4

[0080] 3.1 Antigen preparation

[0081] 3.1.1 Inoculation Dilute chicken inclusion body hepatitis virus SDJ15-8 strain E7 15,000 times with sterilized normal saline, inoculate 7-day-old SPF chicken embryos through the yolk sac, 0.1ml per embryo, seal the holes with wax, and statically place at 37°C. set up for incubation.

[0082]3.1.2 Incubation and observation Discard the chicken embryos that died 48 hours ago, and light the eggs every 6 hours thereafter, collect the chicken embryos that died within 48-120 hours, and refrigerate them at 2-8°C for 12-24 hours.

[0083] 3.1.3 Harvest chicken embryos Disinfect the air cell on the surface of the eggshell with tincture of iodine, remove the eggshell on the air cell by aseptic operation, harvest the allantoic fluid, put it in a sterile container, and mark it as chicken inclusion body hepatitis virus SDJ15-8 strain E8, store a...

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Abstract

The invention discloses an egg yolk antibody of inclusion body hepatitis (IBH) and a preparation method thereof. The egg yolk antibody comprises an IBH virus strain. The virus is avian adenovirus group-I serum 4-type virus and is called SDJ15-8, and is preserved at China Center for Type Culture Collection, Wuhan University, and is assigned the accession number CCTCC No.V201636. The preparation method comprises the steps of: 1) preparing an inactivated vaccine from the IBH virus strain SDJ15-8; 2) performing injection immunization to laying hens with the inactivated vaccine to produce high-immunized eggs; and 3) collecting yolks from the high-immunized eggs, and performing extraction and inactivation in an acidified water-octanoic acid manner to prepare the IBH egg yolk antibody. The egg yolk antibody can effectively prevent and treat the IBH and is more than 82% in curative rate.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to an egg yolk antibody for chicken inclusion body hepatitis and a preparation method thereof. Background technique [0002] Chicken inclusion body hepatitis is characterized by a sudden increase in the death of sick chickens, severe anemia, jaundice, hepatomegaly, hemorrhage and necrosis, and pericardial effusion. The disease occurs mostly in 4-10 week-old chickens, 5-week-old chickens are most susceptible, laying hens are rarely affected, and broiler chickens are more common. The causative agent of the disease belongs to group I of avian adenovirus, which has 12 serotypes. The disease was first reported in the United States in 1951. With the rapid development of my country's chicken breeding industry, the disease has shown a local epidemic trend in broiler chicken breeding areas in my country in recent years. , through the search of Wanfang and V...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/08C07K16/02
CPCC07K16/02C07K16/081C07K2317/11C07K2317/23
Inventor 魏思远
Owner CHONGQING SANJIE ZHONGXIN BIOLOGICAL ENG CO LTD
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