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F1-protein-based indirect immunofluorescence kit for detecting type 4 fowl adenovirus antibody

A poultry adenovirus and immunofluorescence technology, applied in the field of biotechnology detection, to achieve good specificity

Active Publication Date: 2016-10-12
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • F1-protein-based indirect immunofluorescence kit for detecting type 4 fowl adenovirus antibody
  • F1-protein-based indirect immunofluorescence kit for detecting type 4 fowl adenovirus antibody
  • F1-protein-based indirect immunofluorescence kit for detecting type 4 fowl adenovirus antibody

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Experimental program
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Embodiment

[0023] 1. Separation of serum type 4 avian adenovirus: take the liver of a dead chicken suspected of being infected with type 4 adenovirus, grind it and add PBS at a ratio of 1:5 to make a suspension; centrifuge at 5000r / min for 15min, and take the supernatant Add 1000 IU / ml of penicillin and streptomycin, 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 inoculate 96- After 120 hours, the allantoic fluid was collected; after the allantoic fluid was identified as type 4 avian adenovirus, it was stored at -20°C.

[0024] 2. Design and amplify primers containing the pcDNA3.1 linearized vector and the F1 protein gene fragment of serotype 4 avian adenovirus: the specific primer sequences are shown in Table 1, which were synthesized by Suzhou Jinweizhi Biotechnology Co., Ltd.

[0025] Table 1. PCR amplification of linearized pcDNA3.1 and serotype 4 avian adeno...

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Abstract

The invention belongs to the field of biotechnology detection, in particular to an F1-protein-based indirect immunofluorescence kit for detecting a type 4 fowl adenovirus antibody. The kit comprises 293T cells expressing F1 protein, an FITC-labeled goat-anti-chicken antibody, a sample diluent and a washing liquid. The kit has good specificity and not only can be used for epidemiological investigation of serum type 4 fowl adenovirus infection conditions, but also can be used for monitoring the level of a serum type 4 fowl adenovirus antibody of immune chicken flocks and used for evaluating the immunoprotecive antibody level.

Description

technical field [0001] The invention belongs to the field of biotechnology detection, and in particular relates to an indirect immunofluorescence kit for detecting type 4 adenovirus antibody based on F1 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 industry. Viru...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N21/64
CPCG01N21/6486G01N33/5302
Inventor 叶建强梁广成邵红霞王伟康秦爱建万志敏
Owner YANGZHOU UNIV
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