O type foot-and-mouth disease virus artificial recombinant antigen and preparation and application thereof
A foot-and-mouth disease virus and recombinant antigen technology, which is applied in the field of biological preparations, can solve the problems of false positives and missed detection, and achieve the effects of high specificity and sensitivity, easy preparation and operation, and reduced missed detection rate
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Embodiment 2
[0065] The O-type foot-and-mouth disease live vaccine used in Example 2 was purchased from Qianyuanhao Biology Co., Ltd. Cattle O-type FMD virus standard positive serum, guinea pig type A FMD virus standard positive serum, guinea pig type C FMD virus standard positive serum, guinea pig Asia I type FMD virus standard positive serum were donated by Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences. Porcine vesicular disease standard positive serum and bovine viral diarrhea standard positive serum are kept by the Veterinary Diagnostic Center of the Ministry of Agriculture. The positive serum of cattle infected with O-type FMD wild strain was prepared and stored by the Veterinary Diagnostic Center of the Ministry of Agriculture. The positive serum of O-type FMD vaccine strain immunized cattle and the serum of cattle artificially infected with FMD virus were prepared and preserved by Inner Mongolia Biopharmaceutical Factory and the Veterinary Diagnostic...
Embodiment 4
[0066] 10 parts of foot-and-mouth disease negative group bovine serums used in embodiment 4 are provided by Beijing Entry-Exit Inspection and Quarantine Bureau; each 20 parts of foot-and-mouth disease inactivated vaccine immune group bovine serums and foot-and-mouth disease virus artificially infected bovine serums are provided by Inner Mongolia Biopharmaceutical Factory and Ministry of Agriculture veterinarians Jointly prepared and preserved by the diagnostic center; 10 serums from bovines naturally infected with foot-and-mouth disease virus were prepared and preserved by the Veterinary Diagnostic Center of the Ministry of Agriculture;
Embodiment 5
[0067] O-type foot-and-mouth disease natural infection positive serum (20 parts), O-type foot-and-mouth disease artificial infection positive serum (20 parts), foot-and-mouth disease inactivated vaccine immune positive serum, vaccine immune challenge group positive serum (10 parts) and Foot-and-mouth disease negative serum (20 copies) was provided by the Veterinary Diagnostic Center of the Ministry of Agriculture.
[0068] 2. Vector and strain:
[0069] The pGEX-T-Easy vector was purchased from Promega. Dh5a competent cells were purchased from Beijing Quanshijin Biotechnology Co., Ltd. The pGEX-20T vector and the E.coli ER2556 strain were kindly donated by Professor Xia Ningshao from the Tumor and Cell Engineering Laboratory of Xiamen University.
[0070] main reagent
[0071] The total RNA extraction kit is a product of Qiagen. AMV, Rnasin, DNTPs, Taq DNA polymerase, T4 ligase, and pGEM-T-Easy were all purchased from Promega, the gel recovery kit and plasmid extraction ...
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