ELISA reagent kit for testing Asial type foot-and-mouth disease virus
The technology of a foot-and-mouth disease virus and a kit is applied in the field of ELISA kit and preparation for detecting Asia1 type foot-and-mouth disease virus, which can solve the problems of long duration and complicated epidemic situation, and achieve the effect of simple detection process and preparation process.
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[0045] 1. Preparation of monoclonal antibody
[0046] (1) Use the RNeasy Mini Kit (Qiagen) kit to extract the total mRNA from the Asia1 type foot-and-mouth disease virus (note: the strain is stored in the National Foot-and-Mouth Disease Reference Laboratory of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences) as a template, and use the Asia1 type The specific primers of VP1 were reverse-transcribed and PCR-amplified to obtain the VP1 gene of Asia1 type foot-and-mouth disease virus. The primers used in the present invention are:
[0047] Upstream primer 5′-CCG GAATTC GCTTACCAGTGGATGCTCG-3′
[0048] Downstream primer 5-′CCC AAGCTT CGCTAGCTTGAGCAGGTC-3′
[0049] Transform the ligation product into Escherichia coli DH5a competent cells by the usual method, construct the Escherichia coli strain containing the recombinant cloning vector, and use an agarose plate (amp + ) and blue-white-white screened positive clones, and picked white-white clones f...
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