Reagent kit for detecting immunoglobulin m (IgM) AlphaLISA for resisting enterovirus71 capsid protein1
A technology of capsid protein and enterovirus, which is applied in the field of immunological detection of enterovirus 71, can solve problems such as difficult application, and achieve the effect of high sensitivity, good specificity, and less serum consumption
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[0015] 1. Construction and identification of EV71 VP1 recombinant protein prokaryotic expression plasmid
[0016] 1 target fragment amplification
[0017] 1.1 PCR cycle amplification
[0018] Using the genome of strain EV71 / Fuyang.Anhui.CHU / 17.08 / 1 (GenBank No.: EU703812) as a template, perform reverse transcription, and amplify the desired fragment EV71 VP1 1 by conventional polymerase chain reaction (PCR) -297 (GenBank Number: HM579945.1), its nucleotide sequence is shown in Seq ID No.2, and the encoded amino acid sequence is shown in Seq ID No.1.
[0019] The upstream and downstream primers were respectively introduced with Nde I and Xho I restriction sites. The primers were designed using Primer Premier 5 (a primer design software) software. The primers were synthesized by Shanghai Sangon Company. The primer sequences are shown in Table 1.
[0020] Table 1 Primer Sequence
[0021]
[0022] The PCR reaction system is shown in Table 2 (total volume 50 μl):
[0023] Ta...
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