Egg yolk antibody for treating novel reovirus of chicken and preparation method thereof

A technology of reovirus and egg yolk antibody is applied in the field of preventing and treating egg yolk antibody of new chicken reovirus and its preparation, which can solve the problems of inconvenience in use and restricting the development and application of egg yolk antibody of new chicken reovirus, etc. Achieve the effect of good safety, good commercial development prospects, and reduction of impurity content

Active Publication Date: 2018-10-16
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because the disease is reported for the first time in my country, there is no commercialized vaccine and antibody on the market at present, and the egg yolk antibody used for the emergency prevention and early infection treatment of this new type o

Method used

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  • Egg yolk antibody for treating novel reovirus of chicken and preparation method thereof
  • Egg yolk antibody for treating novel reovirus of chicken and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1: Discovery and identification of novel chicken reovirus

[0060] 1. Virus isolation:

[0061] (1) Clinically, the arthritic chicks identified as positive by PCR reaction, the swollen tendon tissue was collected by necropsy and placed in a 15mL centrifuge tube, and 5 times the volume of serum-free DMEM medium was added, and after homogenization, freeze-thaw repeatedly 3 times, shake on the shaker for 1-2min after each thawing, and then proceed to the next freeze-thaw; after freeze-thaw, centrifuge the 15mL centrifuge tube containing the sample in a centrifuge at 4000rpm for 5min; take the supernatant, and use 0.22μm micropore After membrane filtration, set aside;

[0062] (2) According to the proliferation characteristics of reovirus disease, chicken liver cancer cells (LMH) were selected for virus isolation. According to the conventional cell culture method, wait for 25cm 2 When the cells in the cell flask are covered with a monolayer, discard the medium...

Embodiment 2

[0078] Embodiment 2: the preparation of inactivated vaccine

[0079] (1) Virus propagation procedure: inoculate 9-day-old healthy SPF chicken embryos with the novel chicken reovirus strain isolated in Example 1 at a dose of 0.2 ml / piece, discard dead embryos within 24 hours, and collect dead chickens within 24 to 120 hours Embryo allantoic fluid, virus fluid obtained, stored at -20°C.

[0080] (2) Concentration of the virus liquid: Concentrate the harvested virus liquid to 1 / 3 of the original volume with an ultrafiltration concentrator at 2-8°C, and conduct a sterility test according to the appendix of the current "Chinese Veterinary Pharmacopoeia" to ensure aseptic growth , and keep samples for testing the poison price. The virus content per 0.1ml of virus liquid should be ≥10 5.0 ELD 50 , the concentrated allantoic fluid is then inactivated.

[0081] (3) Inactivation: the concentrated virus liquid is introduced into an inactivation tank, and 10% formaldehyde solution is ...

Embodiment 3

[0089] Example 3: Preparation of novel chicken reovirus egg yolk antibody

[0090] 1. Use inactivated vaccines to immunize laying hens:

[0091] The novel chicken reovirus inactivated vaccine prepared in Example 2 is used to immunize laying hens, and the neck of each chicken is subcutaneously injected with 2.0ml of the inactivated vaccine for the first time, and the second immunization is carried out after 14 days, and the neck of each chicken is subcutaneously injected with 2.0ml of the inactivated vaccine. ml of inactivated vaccine, the third immunization is carried out 14 days after the second immunization, and 2.0ml of inactivated vaccine is subcutaneously injected into the neck of each chicken, and booster immunization is carried out 14 days after the third immunization, and 2.0ml of inactivated vaccine is injected subcutaneously into the neck of each chicken 14 days after the booster immunization, the egg yolk was collected to determine the neutralizing antibody titer of...

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Abstract

The invention discloses an egg yolk antibody for treating novel chicken reovirus and a preparation method thereof. The egg yolk antibody comprises an antibody resisting chicken reovirus strain; the preservation number of the chicken reovirus strain is CCTCC NO: V201817. The formed egg yolk antibody is good in safety, and any local or systemic side effect caused by the egg yolk antibody does not occur; infection of the novel chicken reovirus can be effectively prevented and/or treated, and the egg yolk antibody has a good commercial development prospect.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to an egg yolk antibody for preventing and treating novel chicken reovirus and a preparation method thereof. Background technique [0002] Avian orthoreovirus (ARV) belongs to the family Reoviridae and the genus Orthoreovirus. It is a pathogen that causes viral arthritis in chickens. It mainly infects broiler chickens, and chickens with both meat and eggs are also infected. ARV mainly invades the hock joints, toe joints, tibiometatarsal joints and tendons of chickens, causing joint synovitis or tenosynovitis. Diseased chickens usually show symptoms of lameness, squatting, reluctance to walk, loss of appetite or even extinction, resulting in low feed conversion rate, increased number of dead chickens, reduced production efficiency, and caused serious economic losses to the poultry industry. Since the disease was first reported in my country in the mid-1980s, t...

Claims

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Application Information

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IPC IPC(8): C07K16/02C07K16/10A61K39/42A61P31/14
CPCA61K2039/505C07K16/02C07K16/10
Inventor 唐熠刁有祥姜晓宁
Owner SHANDONG AGRICULTURAL UNIVERSITY
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