Aptamer-based enzyme-linked immunosorbent assay method for detecting iridovirus infection of groupers
A technology of enzyme-linked immunosorbent adsorption and nucleic acid aptamer, which is applied in the direction of measuring devices, biological tests, material inspection products, etc., can solve the problems of long time-consuming preparation of antibodies, quality differences between batches, cumbersome operations, etc., and achieve easy operation , short time-consuming, high-sensitivity effect
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[0024] Three nucleic acid aptamers (synthesized by ThermoFisherScientific) for ELISA detection of grouper iridescent virus infection were synthesized, and the sequences were as follows:
[0025] Sequence 1:
[0026] 5'-GACGCTTACTCAGGTGTGACTCGTATGTCCATGGCCGCATATTGGGAAGGTTGGTTTGGACTATGTGGAAGTTCGAAGGACGCAGATGAAGTCTC-3'
[0027] Sequence 2:
[0028] 5'-GACGCTTACTCAGGTGTGACTCGTTTCGTGTTATGCTCCTCTTTATTGTCAGCTGAGTTTCTGCAGTGCGAAGGACGCAGATGAAGTCTC-3'
[0029] Sequence 3:
[0030] 5'-GACGCTTACTCAGGTGTGACTCGTATTCGGGTTATTGCTCCTCTTTATTGTCACCTGGATGTATGATCGTGTAGCGAAGGACGCAGATGAAGTCTC-3'
[0031] The above-mentioned nucleic acid aptamers are modified with biotin tags at the 5' end or 3' end.
[0032] The method for the detection of grouper iridescent virus infection by enzyme-linked immunosorbent assay technology based on nucleic acid aptamer comprises the following steps:
[0033] 1. Refolding of nucleic acid aptamers: biotin-labeled nucleic acid aptamers were denatured at 94°C for 10 mi...
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