JEV (Japanese type B encephalitis virus) nucleic acid assay kit and application thereof
A type of Japanese encephalitis virus and kit technology, applied in the biological field, can solve the problems of disease prevention and control and community detection can not fully play a role, insufficient implementation of immunization, low vaccination rate of Japanese encephalitis, etc., easy to achieve large-scale Promote the application, solve the effect of expensive equipment and quick response
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[0031] 1. Primer design: Download the nucleic acid sequence from Genebank, select the conserved segment of Japanese encephalitis virus, and design and synthesize RPA primers with the following sequence.
[0032] primerA: TGTGTCTTGACCGTGGTTGGAATGGTGGCAGC;
[0033] primerB: GGTTAGCACGACTGTGCTCCCCATACAATGC.
[0034] 2. Probe design:
[0035] 5'-GTACGGGATGCTGGAAAAAACCAAAGCAGAT / THF / TCAAGAGCATGTTTGG / C3-spacer / -3'. The dT near the 5' end (31bp) is labeled with FAM, and the dT near the 3' end (33bp) is labeled with BHQ1.
[0036]Primers and probes were synthesized by Shanghai Sangon Bioengineering Co., Ltd.
[0037] RPA lyophilized reagent, buffer and magnesium salt solution were purchased from TwistAmp of TwistDX Company TM Reagent test kit.
[0038] 3. Reaction system: Add 29.5 μL Rehydration Buffer, 2.1 μL 10 μM PrimerA, 2.1 μL 10 μM Primer B, 0.6 μL 10 μM probe and 10.7 μL double distilled water to the RPA lyophilized reagent. Add 2.5 μL of magnesium acetate to the cap of th...
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