Rapid detection method for ochratoxin A
A technology for ochratoxin and detection method, applied in the field of rapid detection method and detection kit for ochratoxin A, can solve the problems of expensive equipment, high price, complicated operation, etc. Effect
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Embodiment 1
[0030] The kit for detecting ochratoxin A of the present invention at least includes: ochratoxin A nucleic acid aptamer, single-stranded signal probe ssDNA, DNA amplification system, exonuclease, and copper ion reduction detection system. The ochratoxin A nucleic acid aptamer is 5'-GATCGGGTGTGGGTGGCGTAAAGGGAGCATCGGACAC-3'. The single-stranded signal probe ssDNA is 5'-TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTGTGTCCCATGCTCC-3'. Described DNA amplification system comprises buffer solution, and buffer solution is made of Tris-HCl, MgCl 2 , (NH 4 ) 2 SO 4 Composition, deoxymononucleotide triphosphate mixed solution (dNTP) and Phi29 DNA polymerase. The exonuclease is ExoIII exonuclease.
Embodiment 2
[0032] The rapid detection method of ochratoxin A, the specific operation process is as follows:
[0033] Each DNA stock solution was heat-treated at 95°C for 5 min before use, and left at room temperature for 30 min. Then, 40 μL of hybridization buffer solution containing 3.0 μmol ochratoxin A nucleic acid aptamer (Apt) and 3.0 μmol signal probe ssDNA hybridization buffer solution 40 μL were placed in a 2 ml centrifuge tube, and hybridized at 37 ° C for 1 hour to generate Ochratoxin A nucleic acid aptamer-signaling probe hybrid (Apt-ssDNA).
[0034] At 37°C, ochratoxin A with a concentration of 0-50 ng / mL was added to the Apt-ssDNA solution in turn, and ochratoxin A reacted with the ochratoxin A nucleic acid aptamer to generate the nucleic acid aptamer-ochratoxin A, Release of ssDNA. At this time, there are substances such as ssDNA, remaining (unreacted) Apt-ssDNA and nucleic acid aptamer-ochratoxin A in the system.
[0035] Add 10 μL buffer solution (buffer solution compo...
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