Colloidal gold test strip for detecting nervous necrosis virus of grouper and preparation method thereof
A colloidal gold test strip, nerve necrosis technology, which is applied in biological testing, material inspection products, etc., can solve the problems of time-consuming and laborious detection methods for nerve necrosis virus, high detection conditions, and inability to detect in real time, and achieves high sensitivity, specificity, The detection method is simple and the effect of rapid detection
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Embodiment 1
[0068] Such as figure 1 As shown, a colloidal gold test strip for detecting grouper neuronecrosis virus, the colloidal gold test strip includes a base plate 1 and a sample pad 2, a binding pad 3, and nitrocellulose that are sequentially lapped and fixed on the base plate 1 Membrane 4 and absorbent pad 5, the binding pad 3 is coated with a gold-labeled probe, and the gold-labeled probe is a thiol-modified nucleic acid aptamer at the 5' end, and its nucleotide sequence is: 5'-SH - AGCTACTGCTTTGGGGGT(sequence 1);
[0069] The nitrocellulose membrane 4 is provided with a detection line 41 and a quality control line 42, the detection line 41 is fixed with a capture probe, and the quality control line 42 is fixed with a quality control probe:
[0070] The nucleotide sequence of the capture probe is: TTCTTTTATTAGTTGATTT (sequence 2);
[0071] The nucleotide sequence of the quality control probe is: ACCCCCAAAGCAGTAGCT (sequence 3).
[0072] The preparation method of the colloidal g...
Embodiment 2
[0084] A method for detecting grouper neuronecrosis virus using colloidal gold test strip of the present invention, the detection method may further comprise the steps:
[0085] ① Virus incubation: Take 100 μl of phosphate buffered saline (PBS), add capture aptamer (Aptamer1) and amplification aptamer (Aptamer2), make the capture aptamer (Aptamer1) and amplification aptamer (Aptamer2) ) at a concentration of 200nM, and then add grouper neuronecrosis virus (RGNNV) infected cell lysates and incubate for 15min to obtain a mixture containing grouper neuronecrosis virus (RGNNV);
[0086] ②Strand displacement reaction: Add 2 μl of streptavidin (SA)-modified magnetic beads into the mixture, shake for 15 minutes to obtain Aptamer1-RGNNV-Aptamer2-magnetic bead complex, and separate Aptamer1-RGNNV- Aptamer2-magnetic bead complexes were collected, washed three times with phosphate buffered saline (PBST) containing Tween-20, and transferred to centrifuge tubes (EP tubes) for strand displa...
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