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ELISA (Enzyme Linked Immunosorbent Assay) kit for detecting fowl adenovirus antibody based on hexon protein N-terminal conservative area

A technology of avian adenovirus and conserved region, which is applied in the field of biotechnology detection and achieves good specificity.

Inactive Publication Date: 2017-02-22
YANGZHOU UNIV
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  • Abstract
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Problems solved by technology

However, there are no rapid methods and kits for the detection of type 4 FAdV serological antibodies at present.

Method used

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  • ELISA (Enzyme Linked Immunosorbent Assay) kit for detecting fowl adenovirus antibody based on hexon protein N-terminal conservative area
  • ELISA (Enzyme Linked Immunosorbent Assay) kit for detecting fowl adenovirus antibody based on hexon protein N-terminal conservative area
  • ELISA (Enzyme Linked Immunosorbent Assay) kit for detecting fowl adenovirus antibody based on hexon protein N-terminal conservative area

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Embodiment

[0027] 1. Separation of avian adenovirus: take the liver of a dead chicken suspected of avian adenovirus infection, grind it and add PBS at a ratio of 1:5 to make a suspension; centrifuge at 5000r / min for 15min, take the supernatant; add penicillin and strepto Each 1000IU / ml of mycin was reacted at 37°C for 30 minutes; after filtering through a 0.45um microporous filter, 8-day-old SPF chicken embryos were inoculated through the allantoic cavity at a dose of 0.2ml / embryo, and urine was collected 96-120 hours after inoculation. Cystic fluid; allantoic fluid was identified as avian adenovirus and stored at -20°C.

[0028] 2. Design and amplify primers containing the pGEX-6p-1 linearized vector and the N-terminal conserved region of avian adenovirus Hexon protein: the specific primer sequences are shown in Table 1, which were synthesized by Suzhou Jinweizhi Biotechnology Co., Ltd.

[0029] Table 1. PCR amplification of linearized pGEX-6p-1 and avian adenovirus Hexon protein N-term...

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Abstract

The invention belongs to the field of biotechnical detection, and particularly relates to an indirect ELISA (Enzyme Linked Immunosorbent Assay) kit for detecting a fowl adenovirus antibody based on a hexon protein N-terminal conservative area. The kit comprises an ELISA enzyme standard plate coated with the expression product of the hexon protein N-terminal conservative area, an HRP (Horseradish Peroxidase) marked rabbit anti-chicken antibody, a sample diluting solution and a washing solution; the sequence of the hexon protein N-terminal conservative area is as shown by SEQ ID NO. 5. By using the indirect ELISA kit for the fowl adenovirus antibody, which is established by the invention, an antibody resisting a fowl adenovirus can be detected specifically, but an antibody resisting other pathogens cannot be detected. Therefore, the kit has favorable specificity on the fowl adenovirus, and can be used for the epidemiological investigation of the infection status of the fowl adenovirus.

Description

technical field [0001] The invention belongs to the field of biotechnology detection, in particular to an indirect ELISA kit for detecting poultry adenovirus antibody based on the N-terminal conserved region of hexon protein. Background technique [0002] Fowl Adenovirus (Fowl Adenovirus, FAdV) belongs to Adenoviridae, and is divided into 5 species (A-E) and 12 serotypes. Although it has been reported all over the world, FAdV infection generally causes subclinical conditions, while acute infection mainly causes inclusion body hepatitis, pericardial effusion, and gizzard erosion. Since 2013, cases of inclusion body hepatitis and pericardial effusion caused by FAdV in domestic chicken flocks have gradually increased. By 2015, FAdV infection broke out in chicken flocks in several provinces in China. At present, FAdV outbreaks not only occur in 3-4 week-old broiler chickens, but also occur in 10-20 week-old laying hens, causing serious economic losses to the domestic chicken i...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569G01N33/535
CPCG01N33/6857G01N33/535G01N33/56983G01N2333/075
Inventor 叶建强谢泉邵红霞秦爱建田晓彦梁广成王伟康万志敏
Owner YANGZHOU UNIV
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