Method and kit for detecting ochratoxin A

An ochratoxin and a kit technology are applied in the field of a method and kit for detecting ochratoxin A, which can solve the problems of expensive equipment, high price, insufficient sensitivity and the like, and achieve improved detection sensitivity and precision, easy operation, The effect of eliminating distractions

Inactive Publication Date: 2016-05-04
HUNAN UNIV OF SCI & TECH
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Problems solved by technology

[0008] Usually, chromatography is not sensitive enough for the detection of trace fungi in food. Although the liquid-mass spectrometry method has high sensitivity and fast speed, the equipment is expensive, and only professional technicians should be required to perform the detection. The technical requirements are high, and it is not suitable for popularization and However, the method for detecting ochratoxins based on nucleic acid aptamers described in the literature may require the use of expensive labeled (biological materials, etc.) DNA, or have disadvantages such as complicated operations

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  • Method and kit for detecting ochratoxin A

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Embodiment Construction

[0027] The present invention will be described below through specific embodiments.

[0028] The kit for detecting ochratoxin A of the present invention at least includes: an anti-ochratoxin A nucleic acid aptamer, a single-stranded signal probe DNA capable of hybridizing with an anti-ochratoxin A nucleic acid aptamer, a DNA amplification system, an external Dicer, silver ion reduction detection system.

[0029] In one embodiment of the above kit, the anti-ochratoxin A nucleic acid aptamer is GATCGGGTGTGGGTGGCGTAAAGGGAGCATCGGACAC.

[0030] In one embodiment of the above kit, the single-stranded signal probe DNA (Sp) is CCCTTACCCCGGTGTCCCATGCTCCC.

[0031] In one embodiment, the DNA amplification system includes a buffer solution (Tris-HCl, MgCl 2 , (NH 4 ) 2 SO 4 ), dNTPs and Phi29 DNA polymerase.

[0032] In one embodiment, the exonuclease is ExoIII exonuclease.

[0033] Make the anti-ochratoxin A nucleic acid aptamer (Apt) hybridize with the single-stranded signal prob...

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Abstract

The invention relates to a method and kit for detecting ochratoxin A. The method comprises A, carrying out hybridization on an anti-ochratoxin A aptamer (Apt) and a single chain signal probe DNA (Sp) capable of being hybridized with the anti-ochratoxin A aptamer (Apt) so that a hybrid chain is formed, B, contacting the hybrid chain and a sample to be detected, wherein when the sample to be detected contains ochratoxin A, the hybrid chain and ochratoxin A undergo a reaction to produce the single chain signal probe DNA, C, carrying out DNA amplification so that the hybrid chain forms a double-stranded DNA, chopping the double-stranded DNA through an excision enzyme and leaving on the single chain signal probe DNA and D, detecting system fluorescence intensity in a silver ion reduction detection system and determining content of the mycotoxin ochratoxin A. The detection method and kit can eliminate interference and improve detection sensitivity and precision of ochratoxin A.

Description

technical field [0001] The invention belongs to the technical field of nano-biological sensing and biological detection, and relates to a method and a kit for detecting ochratoxin A. Background technique [0002] Ochratoxins include 7 compounds with similar structures, the general structural formula: R1=Cl or H; R2=H, CH3 or C2H5. Among them, ochratoxin A (R1=C1, R2=H) is the most toxic and is most common in moldy grains and feed. [0003] Ochratoxin A is a colorless crystalline compound. Soluble in polar organic solvents and dilute sodium bicarbonate solution. Slightly soluble in water. The melting point of its benzene solvate is 94-96°C, and the melting point of crystals in xylene is 169°C. Optically active [α] D-118 °. Its ultraviolet absorption spectrum varies with pH value and solvent polarity, and the maximum absorption wavelengths in ethanol solution are 213nm and 332nm. It has high chemical stability and thermal stability. Ochratoxin A is produced by various A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/115
CPCC12Q1/6816C12N15/115C12N2310/16C12Q2525/205C12Q2521/319C12Q2563/107
Inventor 易守军冉海宁林雨欢曾云龙邓克勤唐春然
Owner HUNAN UNIV OF SCI & TECH
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