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Preparation methods for duck hepatitis virus immunogen and hyperimmune serum and application of duck hepatitis virus hyperimmune serum

A technology for duck hepatitis virus and hyperimmune serum is applied in the field of animal immunology, which can solve the problems of difficulty and difficulty in obtaining highly specific hyperimmune serum, and achieves a technology with low requirements, simple operation, high specificity and application value. Effect

Inactive Publication Date: 2015-09-23
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The object of the present invention is to provide a kind of preparation method and application of duck hepatitis virus immunogen and hyperimmune serum, aiming at solving the difficulty of obtaining relatively pure virus immunogen and also causing the difficulty of obtaining highly specific hyperimmune serum The problem

Method used

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  • Preparation methods for duck hepatitis virus immunogen and hyperimmune serum and application of duck hepatitis virus hyperimmune serum
  • Preparation methods for duck hepatitis virus immunogen and hyperimmune serum and application of duck hepatitis virus hyperimmune serum
  • Preparation methods for duck hepatitis virus immunogen and hyperimmune serum and application of duck hepatitis virus hyperimmune serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Preparation of Example 1 Serum Type 1 Duck Hepatitis Virus Immunogen

[0031] 1. Virus strain

[0032] Serum Type 1 Duck Hepatitis Virus CH60 strain (Duck Hepatitis A Virus, Duck Hepatitis A Virus), which is a weakened virus strain isolated by the applicant, has applied for a patent (application number 201310011872.3) and was issued in November 2012 On the 20th, the biological material was preserved in the China Center for Typical Culture Collection, the preservation number is CCTCC NO.V201248, and the deposit address is: China Center for Type Culture Collection (Wuhan University) at Luojiashan, Wuchang, Wuhan.

[0033] 2. Experimental embryos

[0034] 9-day-old SPF chicken embryos.

[0035] 3. Virus proliferation

[0036] After diluting the seed virus of the CH60 strain of duck hepatitis virus type 1 5 times with normal saline, add an appropriate amount of double antibody (the final concentrations of penicillin and streptomycin are 1001 U / mL and 100 μg / mL, respectiv...

Embodiment 2

[0041] Preparation of Example 2 Serum Type 3 Duck Hepatitis Virus Immunogen

[0042] 1. Virus strain

[0043]Serum type 3 duck hepatitis virus CH-1 strain (fowl hepavirus belongs to picornaviridae type 3 duck hepatitis A virus, Duck Hepatitis A Vrius type 3, DHAV-3), this virus strain is the virus strain isolated in our laboratory and published in On March 25, 2013, the biological material was preserved at the China Center for Type Culture Collection, the preservation number is CCTCC NO: V201305, and the preservation address is: China Center for Type Culture Collection (Wuhan University) at Luojia Mountain, Wuchang, Wuhan.

[0044] 2. Experimental embryos

[0045] 11-day-old duck embryo.

[0046] 3. Virus proliferation

[0047] After diluting the serum type 3 duck hepatitis virus CH-1 strain 5 times with normal saline, add an appropriate amount of double antibody (the final concentrations of penicillin and streptomycin are 100IU / mL and 100μg / mL, respectively), and pass the ...

Embodiment 3

[0052] Example 3 Preparation of Serum Type 1 Duck Hepatitis Virus Hyperimmune Serum

[0053] 1. Immunogen

[0054] The CH60 strain immunogen of serotype 1 duck hepatitis virus prepared in Example 1.

[0055] 2. Experimental animals

[0056] 5 healthy rabbits.

[0057] 3. Adjuvant

[0058] Complete Freund's adjuvant was purchased from Sigma. Mix paraffin oil and lanolin at a ratio of 1:5 by volume, mix them evenly with ultrasonic waves, and sterilize them at high temperature to make incomplete Freund's adjuvant.

[0059] 4. Animal immunity

[0060] (1) Immunization for the first time: Dilute the immunogen 2ml (estimated to be 1500 μg / ml×2ml=3mg) of the serum type 1 duck hepatitis virus CH60 strain immunogen prepared in Example 1 to 3ml with normal saline, and use the purchased Sigma company 3ml of Shi's complete adjuvant was emulsified according to the conventional method until it became a uniform milky white "water-in-oil" state (take a drop of the liquid and place it on...

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Abstract

The invention provides preparation methods for duck hepatitis virus immunogen and hyperimmune serum and application of the duck hepatitis virus hyperimmune serum. According to the preparation methods and the application of the duck hepatitis virus hyperimmune serum, the duck hepatitis virus immunogen is obtained through inoculating a serum 1 type duck hepatitis virus CH60 strain DHAV-1 (Duck Hepatitis A Virus type 1) or a serum 3 type duck hepatitis virus CH1 strain DHAV-3 (Duck Hepatitis A Virus type 3) to an allantoic cavity of a chick embryo of 9-10 days old or a duck embryo of 10-12 days old and carrying out proliferation and treatment, and the hyperimmune serum is obtained through mixing the duck hepatitis virus immunogen with a Freund's complete adjuvant or Freund's incomplete adjuvant to prepare solutions of different concentrations, carrying out repeated immunization on immune animals and then sampling and collecting blood and can be applied to the diagnosis and detection on a duck hepatitis virus. The preparation methods provided by the invention have the advantages that the conditional requirements are low, the operation is simple, and the obtained immunogen can meet the requirements on the preparation of specific antiserum.

Description

technical field [0001] The invention belongs to the field of animal immunology, and in particular relates to a preparation method and application of duck hepatitis virus immunogen and hyperimmune serum. Background technique [0002] Duck Viral Hepatitis (DVH) is a highly fatal infectious disease caused by Duck Hepatitis Virus (DHV), characterized by acute onset, short course of disease, high morbidity and mortality. Duck viral hepatitis is divided into three independent serologically unrelated serotypes: type I, type II and type III. Wherein, serum type 1 duck hepatitis virus (DHV-I) is also called duck hepatitis A virus (Duck hepatitis A virus, DHAV), is the main duck hepatitis virus popular in my country at present, has caused huge economic loss to my country's duck industry , threatening the healthy development of the duck industry. [0003] Duck hepatitis A virus has been found to have three genotypes, namely DHAV-1, DHAV-2 and DHAV-3. According to reports, DHAV-1 is cu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C07K16/06G01N33/569G01N33/531
Inventor 程安春汪铭书朱德康杨乔陈孝跃
Owner SICHUAN AGRI UNIV
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