Method for detecting T-2 toxin

A detection method, T-2 technology, applied in the direction of measuring devices, biological tests, material inspection products, etc., can solve problems such as complex procedures, inability to meet the requirements of T-2 real-time and rapid detection, time-consuming and labor-intensive, etc., to achieve high selection sexual effect

Pending Publication Date: 2021-02-05
GANSU AGRI UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, methods that can effectively detect T-2 include high-performance liquid chromatography, liquid chromatography-mass spectrometry, ultra-high performance liquid chromatography-tandem mass spectrometry, and immunoassay-based detection methods, but these methods need to rely on expensive instruments, and actual detection The process is complicated, time-consuming and labor-intensive, and requires high professionalism of operators, which cannot meet the real-time and rapid detection requirements of T-2.

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  • Method for detecting T-2 toxin
  • Method for detecting T-2 toxin
  • Method for detecting T-2 toxin

Examples

Experimental program
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Embodiment 1

[0034] (1) Take T-2 standard solutions of different known concentrations as samples and perform the following operations to obtain corresponding standard reaction solutions: In a 1.5mL centrifuge tube, add 50μL, 1μM single-stranded DNA aptamer and 50μL sample Add 157 μL of 10 mM, pH 7.0 MOPS buffer, shake and mix, incubate at room temperature for 2 h, then add 200 μL of 10 nM AuNPs and incubate for 30 min, then add 43 μL of 1M NaCl solution and incubate for 10 min to obtain 500 μL of standard reaction solution;

[0035] (2) Each standard reaction solution is transferred to a 96-well plate, and the A620 / A520 standard value corresponding to each concentration T-2 standard solution is measured, and the curve of absorbance linear change drawn according to each A620 / A520 standard value, in this curve, In the concentration range of 0.21435~10717.5nM, the concentration of T-2 standard solution has a linear relationship with the value of A620 / A520, and the linear equation is obtained: ...

Embodiment 2

[0039] (1) Take T-2 standard solutions of different known concentrations as samples and perform the following operations to obtain corresponding standard reaction solutions: In a 1.5mL centrifuge tube, add 40μL, 1μM single-stranded DNA aptamer and 40μL sample Add 180 μL of 10 mM, pH 7.0 MOPS buffer, shake and mix, incubate at room temperature for 1 hour, then add 200 μL of 10 nM AuNPs and incubate for 20 minutes, then add 40 μL of 1M NaCl solution and incubate for 5 minutes to obtain 500 μL of standard reaction solution;

[0040] (2) Each standard reaction solution is transferred to a 96-well plate, and the A620 / A520 standard value corresponding to each concentration T-2 standard solution is measured, and the curve of absorbance linear change drawn according to each A620 / A520 standard value, in this curve, In the concentration range of 0.21435~10717.5nM, the concentration of T-2 standard solution has a linear relationship with the A620 / A520 value, and the linear equation is obt...

Embodiment 3

[0044] (1) Use T-2 standard solutions of different known concentrations as samples and perform the following operations to obtain corresponding standard reaction solutions: In a 1.5mL centrifuge tube, add 60μL, 1μM single-stranded DNA aptamer and 60μL sample Add 130 μL of 10 mM, pH 7.0 MOPS buffer, shake and mix, incubate at room temperature for 1 h, then add 200 μL of 10 nM AuNPs and incubate for 40 min, then add 50 μL of 1M NaCl solution and incubate for 20 min to obtain 500 μL of standard reaction solution;

[0045] (2) Each standard reaction solution is transferred to a 96-well plate, and the A620 / A520 standard value corresponding to each concentration T-2 standard solution is measured, and the curve of absorbance linear change drawn according to each A620 / A520 standard value, in this curve, In the concentration range of 0.21435~10717.5nM, the concentration of T-2 standard solution has a linear relationship with the A620 / A520 value, and the linear equation is obtained;

[...

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Abstract

The invention discloses a visual detection method of T-2 toxin. The method comprises the following steps: oscillating and uniformly mixing a single-stranded DNA aptamer, a sample and an MOPS buffer solution, performing incubating, adding AuNPs, incubating, adding a NaCl solution, performing incubating, determining the A620 / A520 value of the reaction solution to be detected, and substituting the A620 / A520 value into a linear equation constructed by a T-2 standard solution to calculate the toxin concentration in the sample. The lowest detection limit of the method on the T-2 toxin is 0.124 nM (57.8 pg / mL), the method has obvious advantages in the aspects of operation process, result and the like; in addition, the method has high selectivity on the detection of the T-2 toxin, and under the same condition, the method disclosed by the invention is not influenced by other mycotoxins including aflatoxin B1, ochratoxin A, zearalenone and fumonisins B1.

Description

technical field [0001] The invention belongs to the technical field of trace toxin detection, and in particular relates to a detection method of T-2 toxin. Background technique [0002] T-2 toxin is a highly toxic fungal secondary metabolite widely distributed in wheat, corn, grain and rice and their products. T-2 toxin can inhibit DNA, RNA, protein synthesis and mitochondrial function in vivo and in vitro, and has teratogenic, carcinogenic and mutagenic effects. According to the recommendations of the European Food Safety Committee, the daily allowable intake and acute reference dose of T-2 are 0.02 μg / kg and 0.3 μg / kg respectively, which requires a highly sensitive and accurate T-2 detection method, In order to correctly assess the risks people face. [0003] At present, methods that can effectively detect T-2 include high-performance liquid chromatography, liquid chromatography-mass spectrometry, ultra-high performance liquid chromatography-tandem mass spectrometry, and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N21/31
CPCG01N33/5308G01N21/31
Inventor 张紊玮王艳玲毕阳
Owner GANSU AGRI UNIV
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