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Indirect ELISA (Enzyme-linked Immuno Sorbent Assay) kit for detecting blood serum 8 type avian adenovirus antibody based on spike glycoprotein F

A technology of poultry adenovirus and spike protein, which is applied in the field of biotechnology detection and achieves good specificity

Active Publication Date: 2018-06-01
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no rapid method and kit for FAdV-8 serological antibody detection at present

Method used

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  • Indirect ELISA (Enzyme-linked Immuno Sorbent Assay) kit for detecting blood serum 8 type avian adenovirus antibody based on spike glycoprotein F
  • Indirect ELISA (Enzyme-linked Immuno Sorbent Assay) kit for detecting blood serum 8 type avian adenovirus antibody based on spike glycoprotein F
  • Indirect ELISA (Enzyme-linked Immuno Sorbent Assay) kit for detecting blood serum 8 type avian adenovirus antibody based on spike glycoprotein F

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The indirect ELISA kit of the present invention can be obtained through the following steps:

[0042] (1) Isolation of FAdV-8 avian adenovirus:

[0043] Take (suspected) liver of dead chicken infected by avian adenovirus, grind it and add PBST at a ratio of 1:5 to make a suspension; centrifuge at 5000r / min for 15min, take the supernatant; ml, react at 37°C for 30 minutes; filter through a 0.45um microporous filter, inoculate 8-day-old SPF chicken embryos through the allantoic cavity at a dose of 0.2ml / embryo, and collect allantoic fluid 96-120 hours after inoculation; allantoic fluid After the solution was identified as FAdV-8 avian adenovirus, it was stored at -20°C.

[0044] (2) Preparation of avian adenovirus genome:

[0045] Get 400uL supernatant of FAdV-8 isolated strain virus (serum type 8 avian adenovirus) in a 1.5mL finger tube, add 400uL cell lysate (50mmol / LTris-HCl pH 8.0, 20mmol / L EDTA pH 8.0, 2% SDS and Proteinase K), mix well and place in a water bath a...

Embodiment 2

[0060] Infection sample detection effect

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Abstract

The invention discloses an indirect ELISA (Enzyme-linked Immuno Sorbent Assay) kit for detecting a blood serum 8 type avian adenovirus antibody based on spike glycoprotein F. The indirect ELISA kit comprises an ELISA enzyme-linked plate which is coated with expression protein of blood serum 8 type avian adenovirus spike glycoprotein gene F, positive and negative controls, an HRP (Horseradish Peroxidase) labeled rabbit anti-chicken secondary antibody, a diluting solution, a color developing solution, a washing solution and a stopping solution. The ELISA kit disclosed by the invention can be used for specifically detecting a FAdV-8 antibody and does not react with anti-influenza virus blood serum, anti-Marek's virus blood serum, anti-Newcastle disease virus blood serum, anti-chicken infectious anemia virus blood serum, reticuloendotheliosis virus, ant-blood serum 4 type avian adenovirus blood serum, anti-egg drop syndrome virus blood serum and SFP chicken blood serum. Therefore, the kitdisclosed by the invention has good FAdV-8 specificity and can be used for FAdV-8 infection and immunization status epidemiological surveys.

Description

technical field [0001] The invention belongs to the field of biotechnology detection, in particular to an indirect ELISA kit for detecting serum 8 avian adenovirus antibody based on spike protein F. Background technique [0002] Fowl Adenovirus (Fowl Adenovirus, FAdV) belongs to the family 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, the outbreak of FAdV in China not only occurs in 3-4 week-old broilers, but also occurs in 10-20 week-old laying hens, causing serious economic losses to the chicken industry. Virus i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543G01N33/535G01N21/78G01N21/31
CPCG01N21/31G01N21/78G01N33/535G01N33/543G01N33/56983G01N2333/075
Inventor 叶建强路浩邵红霞秦爱建王伟康田晓彦
Owner YANGZHOU UNIV
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