Deer epidemic hemorrhage competitive enzyme-linked immunosorbent assay kit and preparation method and use thereof
A deer epidemic hemorrhage and enzyme-linked immunosorbent assay technology, applied in the biological field, can solve problems such as economic loss of EHDV, and achieve the effect of high sensitivity and strong specificity
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[0046] The preparation method of the deer epidemic hemorrhagic disease virus antibody competition enzyme-linked immunosorbent assay kit of the present invention includes but is not limited to the following examples. The concrete steps of this method are:
[0047] 1. Preparation of deer epidemic hemorrhagic disease virus ELISA coated antigen: BHK-21 cells grow into a single layer and inoculate with deer epidemic hemorrhagic disease virus EHDV, and react at 37°C for 1 hour. Serum-free basal medium was added to continue culturing, and the virus was harvested when the cytopathic pathology (CPE) reached more than 75% after 3 to 4 days. The harvested virus liquid was repeatedly frozen and thawed 3 times, centrifuged at 8000 rpm for 30 minutes, the supernatant was taken, and the virus was inactivated by adding formaldehyde with a final concentration of 1 / 3000. Ultracentrifuge at 28,000 rpm and 4°C for 3 hours. The precipitate was dissolved with 1 / 100 of the original volume of PBS (...
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