A kind of apple virus detection method and detection kit used therefor
A technology for detecting kits and apple viruses, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, and microorganism measurement/inspection. The effect of high detection efficiency and easy modification
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Embodiment 1
[0055] 1. Synthesis of nano-gold apple stem groove virus nucleic acid probe:
[0056] The sulfhydryl-labeled capture probe DNA1 and the fluorophore-labeled signal probe DNA2 were hybridized in the hybridization buffer at a concentration ratio of 1:1.1, and then the hybridized double-stranded DNA solution was mixed with the nano-gold solution at a ratio of 298:1. The concentration ratio was mixed, and the double-stranded DNA was connected to the nano-gold surface with a particle size of 14nm and a concentration of 3nmol / L through the Au-S bond, and the nano-gold apple stem groove virus nucleic acid probe solution was synthesized.
[0057] 2. Detect the apple virus to be tested
[0058] Take 100 μL of nano-gold stem groove virus nucleic acid probe solution and add 200 μL of the prepared detection buffer to mix, the final concentration of nano-gold is 1nmol / L, add artificially synthesized stem groove apple virus oligonucleotide T1 sequence powder to be detected, Check it out. B...
Embodiment 2
[0061] 1. Synthesis of nano-golden apple mosaic virus nucleic acid probe:
[0062] The thiol-labeled capture probe DNA3 and the fluorophore-labeled signal probe DNA4 were hybridized in the hybridization buffer at a concentration ratio of 1:1.2, and then the hybridized double-stranded DNA solution was mixed with the nano-gold solution at 300:1 The concentration ratio was mixed, and the double-stranded DNA was connected to the nano-gold surface with a particle size of 13nm and a concentration of 3nmol / L through the Au-S bond to synthesize a nano-gold apple mosaic virus nucleic acid probe solution.
[0063] 2. Detect the apple virus to be tested
[0064] Take 100 μL of nano-gold mosaic virus nucleic acid probe solution and add 200 μL of the prepared detection buffer to mix, the final concentration of nano-gold is 1 nmol / L, add artificially synthesized mosaic apple virus oligonucleotide T2 sequence powder to be tested, and Check it out. Because it contains mosaic apple virus, th...
Embodiment 3
[0067] The implementation steps of the apple virus detection method of the present embodiment are basically the same as in Example 2, the difference being that the gold nanoparticle diameter is 15nm, the concentration of the capture probe DNA3 labeled with sulfhydryl group and the signal probe DNA4 labeled with fluorophore according to 1:1.3 Compared with hybridization in the hybridization buffer, when the apple virus to be tested is detected, the artificially synthesized T3 sequence with a single base mismatch between the target apple mosaic virus oligonucleotide and the target apple mosaic virus oligonucleotide is detected. The sequence to be detected has a single base mismatch with the mosaic apple virus nucleotide, and the sequence length is long, but it can only partially complement the sulfhydryl-labeled DNA3 capture probe, and the fluorophore-labeled signal probe can only be a small Some of them are replaced, most of the signal probes are close to the gold nanoparticles,...
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