Detection method and detection kit for porcine type A foot-and-mouth disease virus specific IgA antibody
A technology of foot-and-mouth disease virus and detection kit, which is applied to measuring devices, instruments, scientific instruments, etc., to achieve the effects of low manpower consumption, easy operation, and low stress
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[0038] Preparation method of the kit:
[0039]1. Preparation of coating antigen and selection of optimal coating concentration:
[0040] The 636bP fragment of the A-type FMDV-VP1 protein was cloned into the pET-30a (+) prokaryotic expression vector. After digestion and sequencing, it was transformed into E.coli BL-21 (E3), and the VP1 protein was induced by IPTG. Proteins were purified using the Ni-Agarose His-Tag Protein Purification Kit. The purified product expressed by the prokaryotic expression system was serially diluted with carbonate buffer 7μg / mL, 3.5μg / mL, 1.75μg / mL, 0.88μg / mL, 0.44μg / mL, 0.22μg / mL, each Two columns were coated with each concentration, 100 μL / well was added to a 96-well microtiter plate, and coated overnight at 4°C. The results showed that when the coating antigen concentration was 3.5μg / mL, the positive value OD450 value was greater than 1.0, and the value of P / N was the largest, so this concentration was determined to be the optimal coating conce...
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