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Method for rapidly detecting T-2 toxin

A technology of T-2 and toxin, which is applied in the direction of material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of easy inactivation, expensive instruments, difficult to popularize, etc., and achieve the effect of eliminating interference and improving detection sensitivity and accuracy

Inactive Publication Date: 2016-05-04
HUNAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, each of these methods has its own disadvantages: such as chromatography-mass spectrometry, not only the instrument is expensive, but also needs specialized technical personnel to be competent, so it is difficult to popularize; immunoassay reagents are expensive and easy to inactivate; The method for measuring T-2 toxin by formula reaction needs to use expensive DNA through labeling, etc.

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  • Method for rapidly detecting T-2 toxin

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Embodiment 1

[0027] The kit for detecting T-2 toxin of the present invention at least includes: T-2 toxin nucleic acid aptamer, single-stranded signal probe ssDNA, DNA amplification system, exonuclease, copper ion reduction detection system. The T-2 toxin nucleic acid aptamer is 5'-CAGCTCAGAAGCTTGATCCTGTATATCAAGCATCGCGTGTTTACACATGCGAGAGGTGAAGACTCGCA-3'. The single-stranded signal probe ssDNA is 5'-TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTGCGAGTCTTCACC-3'. Described DNA amplification system comprises buffer solution (Tris-HCl, MgCl 2 , (NH 4 ) 2 SO 4 ), dNTPs and Phi29 DNA polymerase. The exonuclease is ExoIII exonuclease.

Embodiment 2

[0029] A method for rapid detection of T-2 toxin, the specific operation process is as follows:

[0030] Each DNA stock solution was heat-treated at 95°C for 5 min before use, and left at room temperature for 30 min. Then, take 40 μL of the hybridization buffer solution containing 3.0 μmol of the T-2 toxin aptamer Apt and 40 μL of the hybridization buffer solution of 3.0 μmol of the signal probe ssDNA in a 2 ml centrifuge tube, and hybridize at 37°C for 1 hour to generate T-2 Toxin nucleic acid aptamer-signaling probe hybrid (Apt-ssDNA).

[0031] At 37°C, add the T-2 toxin with a concentration of 0~50ng / mL to the Apt-ssDNA solution in turn, and the T-2 toxin reacts with the T-2 toxin nucleic acid aptamer to generate the nucleic acid aptamer-T-2 toxin, releasing ssDNA. At this time, there are substances such as ssDNA, residual (unreacted) Apt-ssDNA and aptamer-T-2 toxin in the system.

[0032] Add 10 μL buffer solution (buffer solution composition is 50mM Tris-HCl, 10mM MgCl...

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Abstract

The invention relates to a method for rapidly detecting a T-2 toxin. The method comprises the following steps that A, a T-2 toxin aptamer and single-strand signal probe DNA are hybridized to form a hybrid strand; B, the hybrid strand makes contact with a sample to be detected, and the hybrid strand reacts with the T-2 toxin to release out the single-strand signal probe DNA when T-2 toxin exists in the sample to be detected; C, the hybrid strand is made to form two-strand DNA by means of DNA amplification, then excision enzyme is used for hydrolyzing the two-strand DNA into mononucleotide, and at the moment, the single-strand signal probe DNA is left in the system; D, under the induction of the single-strand DNA, copper ions are restored to generate near infrared fluorescent copper nano particles; the system fluorescence intensity is detected, and therefore the content of the T-a toxin in the sample to be detected is detected. The method has the advantages of being high in sensitivity, easy and fast to operate, low in cost and the like.

Description

technical field [0001] The invention belongs to the technical field of nanometer biosensing and biodetection, in particular, it provides a method for rapidly detecting T-2 toxin. Background technique [0002] T-2 toxin is mainly one of the trichothecene compounds produced by fungi such as Fusarium tritinarum. It is widely distributed in nature and is the main toxin that commonly pollutes field crops and stored grains. It is harmful to humans and animals. The Food and Agriculture Organization of the United Nations (FAQ) and the World Health Organization (WHO) regard this type of toxin as the most dangerous source of food contamination that occurs naturally. T-2 toxin is very stable at room temperature, and its toxicity cannot be destroyed by placing it for 6-7 years or heating it to 100-120°C for 1 hour. So far, there is no specific prevention and treatment method for T-2 toxin poisoning, and the only effective prevention method is to avoid or reduce exposure. Therefore, th...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/64
Inventor 易守军曾秀颜邓克勤黄昊文唐春然夏晓东
Owner HUNAN UNIV OF SCI & TECH