Kit and preparation method for detecting foot-and-mouth disease type a, type o and asia 1 virus
A detection kit and a technology for foot-and-mouth disease virus, which are applied to primers of O-type and Asia1-type viruses, detect the field of foot-and-mouth disease type A, and can solve problems such as inability to distinguish between serotypes
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[0038] The present invention will be explained in detail below in conjunction with the examples.
[0039] 1. Preparation of Sequences
[0040] According to the GeXP primer design requirements and the genomes of FMDV-A, FMDV-O, and FMDV-Asia1 published by NCBI, select conserved regions to design specific primers, and form specific chimeric primers by adding universal primers. The universal primer sequence belongs to A non-biologically derived nucleotide sequence, in addition to synthesizing a universal primer sequence, and adding a Cy5 fluorescent label to the 5' end of the upstream universal primer.
[0041] 2. Viral Genome Extraction
[0042] After the monolayer cells grow to more than 80% confluent, inoculate the virus, incubate at 37°C for 30 minutes, and discard the venom. Add cell maintenance solution, 37°C 5% CO 2 Cultivate, observe cytopathic changes (CPE) regularly, and collect the poison after more than 90% of the cells appear CPE. The virus was repeatedly frozen...
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