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Duck adenovirus type 3 strain, duck adenovirus egg yolk antibody as well as preparation method and application of duck adenovirus type 3 strain and duck adenovirus egg yolk antibody

A technology of egg yolk antibody and adenovirus, applied in the field of bioengineering, can solve the problem that there is no particularly effective means for the prevention and control of adenovirus type 3

Active Publication Date: 2021-08-20
重庆永健生物技术有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no particularly effective method for the prevention and control of adenovirus type 3. Egg yolk antibody can be used for the treatment and prevention of specific diseases of livestock and poultry, especially for the specific treatment of viral diseases. It is a convenient, efficient and non-toxic Non-antibiotic biological poultry medicine with side effects has been widely recognized and applied clinically, and at the same time, it can avoid virus contamination that may be caused by vaccines
There is no commercial yolk antibody product on the market, so there is an urgent need for a yolk antibody product that can be used to prevent and treat adenovirus type 3

Method used

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  • Duck adenovirus type 3 strain, duck adenovirus egg yolk antibody as well as preparation method and application of duck adenovirus type 3 strain and duck adenovirus egg yolk antibody
  • Duck adenovirus type 3 strain, duck adenovirus egg yolk antibody as well as preparation method and application of duck adenovirus type 3 strain and duck adenovirus egg yolk antibody
  • Duck adenovirus type 3 strain, duck adenovirus egg yolk antibody as well as preparation method and application of duck adenovirus type 3 strain and duck adenovirus egg yolk antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] 1. Test material

[0038] The disease material came from a duck farm suspected to be infected with adenovirus in Guangdong Province; LMH cells were preserved by Chongqing Yongjian Company.

[0039] 2. Test method

[0040] (1) Collection and processing of disease materials

[0041] The liver and spleen were collected from muscovy ducks suspected of being infected with adenovirus. After repeated freezing and thawing, 1% penicillin-streptomycin was added and then distributed into 1.5 mL sterile EP tubes. Filtered with a 0.22 μm bacterial filter, the Store at -80°C for later use.

[0042] (2) Virus detection

[0043] Total RNA was extracted with a commercial DNA / RNA extraction kit, reverse-transcribed to obtain a cDNA template, PCR amplification was performed using a pair of specific primers for the DAdv gene, and PCR products were detected by 1% agarose gel electrophoresis.

[0044] Table 1 identifies the primers of the SVA gene

[0045]

[0046] (3) HEXON sequenci...

Embodiment 2

[0093] The preparation of embodiment 2 duck adenovirus yolk antibody

[0094] 1. Immunization of laying hens

[0095] (1) Basic immunization chickens were subcutaneously injected with 1.0 mL of immune antigen per chicken.

[0096] (2) Booster immunization 14 days after the basic immunization, chickens were subcutaneously injected with 2.0 mL of immune antigen per chicken.

[0097] (3) Booster immunization 21 days after the booster immunization, chickens were subcutaneously injected with 2.0 mL of immune antigen per chicken.

[0098] (4) Maintenance of immunity When the yolk neutralizing antibody titer of goose Astrovirus in highly immune eggs is critical at 1:512, maintain vaccination once, and subcutaneously inject 2.0 mL of immune antigen per chicken.

[0099] 2. High-free egg collection

[0100] Seven days after the booster immunization, eggs were sampled to detect duck adenovirus virus yolk neutralizing antibody titer ≥ 1:512 as qualified. Collect the eggs and store th...

Embodiment 3

[0117] 1. Virus detection

[0118] RNA was extracted from tissue disease materials suspected of duck adenovirus infection, and PCR amplification was performed using cDNA obtained by reverse transcription as a template. The reaction conditions were: pre-denaturation at 94°C for 5 minutes; denaturation at 94°C for 30s, annealing at 55°C for 30s; extension at 72°C 30s; 30 cycles; 10min at 72°C. Electrophoresis of PCR products on 1% agarose gel for detection results figure 1 , it can be seen from the results that a band of about 633bp was amplified, which was consistent with the size of the expected amplified fragment. Confirmed to be duck adenovirus.

[0119] 2. Virus HEXON gene sequencing and sequence analysis

[0120] The DNAstar software was used to splice the sequencing results of the two overlapping fragments to obtain the duck adenovirus HEXON gene sequence. The results of the HEXON gene nucleotide alignment were analyzed. The nucleotide homology between the isolated H...

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Abstract

The invention discloses a duck adenovirus type 3 strain, a duck adenovirus egg yolk antibody as well as a preparation method and application of the duck adenovirus type 3 strain and the duck adenovirus egg yolk antibody. The duck adenovirus type 3 strain is a duck adenovirus type 3 YJ strain and is preserved in the China General Microbiological Culture Collection Center on November 18, 2020, and the preservation number of the duck adenovirus type 3 strain is CGMCC No.21093; and the nucleotide sequence of the HEXON gene of the duck adenovirus 3 type YJ strain is as shown in SEQ ID NO. 1. The adenovirus 3 type YJ strain is obtained through separation, and the egg yolk antibody which can be used for treating the duck adenovirus 3 type virus and is efficient, safe and free of toxic and side effects is prepared.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a duck adenovirus type 3 bacterial strain, duck adenovirus yolk antibody and a preparation method and application thereof. Background technique [0002] Adenovirus is a linear double-stranded DNA virus without envelope, with a diameter of 70-110nm and icosahedral symmetry. Adenovirion particles consist of 252 capsomers, including 240 non-apical capsomers (hexons) and 12 apical capsomers (pentons). Each penton base has 1 to 2 fibrils called fibrils, and the length of the fibrils is related to the antigenicity of the virus. There are mainly 3 groups of adenoviruses that infect poultry. The representative strain of group I is chicken embryo lethal orphan virus. At present, 12 serotypes have been isolated from chickens, 2 serotypes from turkeys, 4 serotypes from geese, and 3 serotypes from ducks. Type II; group II mainly includes turkey hemorrhagic enteritis viru...

Claims

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Application Information

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IPC IPC(8): C12N7/00C07K16/08A61K39/395A61P31/20
CPCC12N7/00C07K16/081A61P31/20C12N2710/10221Y02A50/30
Inventor 李来旭李睿刘鑫莹何鸿章张强周宇明月月李阁张晏胡芸
Owner 重庆永健生物技术有限责任公司
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