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
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[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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