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A method for rapid and sensitive detection of tetrodotoxin ttx

A technology for sensitive detection of tetrodotoxin, applied in measurement devices, material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of poor selectivity, low reaction sensitivity, long measurement time, etc., and achieve low cost, high sensitivity, and good selectivity. Effect

Active Publication Date: 2021-09-28
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] In order to solve the problems of long measurement time, low reaction sensitivity, poor selectivity, and high measurement cost in existing measurement methods, the present invention provides a method for fast and sensitive detection of tetrodotoxin TTX, using berberine as a fluorescent probe for detection TTX, in the presence of the detection substance TTX, induces a change in the structure of the aptamer and combines with a fluorescent probe to achieve a change in the fluorescent signal

Method used

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  • A method for rapid and sensitive detection of tetrodotoxin ttx
  • A method for rapid and sensitive detection of tetrodotoxin ttx
  • A method for rapid and sensitive detection of tetrodotoxin ttx

Examples

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Embodiment 1

[0027] Example 1: A method for rapid and sensitive detection of tetrodotoxin TTX, adding tetrodotoxin nucleic acid aptamer and berberine to phosphate buffer solution, using berberine as fluorescent probe and tetrodotoxin nucleic acid aptamer as recognition Unit, adding the analyte tetrodotoxin induces the structure change of the nucleic acid aptamer, the binding of the nucleic acid aptamer and the fluorescent probe berberine is affected, causing the change of the fluorescence signal of the system, detecting the fluorescence intensity value of the system, and calculating the analyte content. Tetrodotoxin nucleic acid aptamer is: TTX-aptamer: 5'-ttt tta aag tgt gcc cac gga gcc gac agg-3'.

[0028] Specific steps are as follows:

[0029] (1) Make a standard curve: Add magnesium chloride at a concentration of 1.0 M, nucleic acid aptamer at a concentration of 100 μM and fluorescent probe at a concentration of 2.0 mM into a phosphate buffer solution with a concentration of 1 mM as ...

experiment example 1

[0032] Experimental Example 1: Investigate the optimum pH of the system. Select five buffer solutions with different pH (pH 6.5-8.5), add magnesium chloride, nucleic acid aptamer, TTX respectively, and finally add fluorescent probe, mix well and let stand for 5-10 min, measure the fluorescence at 530 nm wavelength strength. Prepare blank solution in the same way. see results figure 2 . Depend on figure 2 It can be seen that when the pH value of the system is 7.5, (F-F 0 ) / F 0 The value changes the most, and the pH of the system is planned to be 7.5.

experiment example 2

[0033] Experimental Example 2: Investigation of the concentration of the salt solution. Select buffer solutions with different salt concentrations (0.5 mM-3.0 mM), add nucleic acid aptamers, TTX, and finally add fluorescent probes, mix well and let stand for 5-10 min to measure the fluorescence intensity of the system. Prepare blank solution in the same way. see results image 3 . Depend on image 3 It can be seen that when the concentration of magnesium chloride in the solution is 2.0-2.5 mM, (F-F 0 ) / F 0 The value changes the most, and the concentration of magnesium chloride in the system is planned to be 2.0-2.5 mM.

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Abstract

The invention belongs to the technical field of biological detection. In order to solve the problems of long measurement time, low reaction sensitivity, poor selectivity and high measurement cost in existing measurement methods, a method for rapidly and sensitively detecting tetrodotoxin TTX is provided. Tetrodotoxin Nucleic acid aptamer and berberine are added to the phosphate buffer solution, berberine is used as a fluorescent probe, and tetrodotoxin nucleic acid aptamer is used as a recognition unit. After adding tetrodotoxin to induce a structural change of the nucleic acid aptamer, the The combination of the aptamer and the fluorescent probe berberine is affected, which causes the change of the fluorescence signal of the system, the fluorescence intensity value of the system is detected, and the content of the analyte is calculated. It is simple, low in cost, high in sensitivity and good in selectivity, and can be used for the detection of actual biological samples and has potential application value.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and specifically relates to a method for rapidly and sensitively detecting tetrodotoxin TTX, using berberine as a fluorescent probe to detect TTX, and inducing a change in the structure of the aptamer to combine with the fluorescent probe when the detection substance TTX exists , to achieve a change in the fluorescent signal. Background technique [0002] Tetrodotoxin (TTX) is a highly toxic small molecule alkaloid. Its molecular formula is C 11 h 17 o 8 N 3 , with a molecular weight of 319, the substance is chemically stable, stable to heat, sunlight, enzymes, salts and acids, and easily soluble in dilute acetic acid. The chemical structural formula of TTX is as follows figure 1 As shown, it is an amino perhydroquinazoline compound, which was first isolated from puffer fish. In recent years, it has also been found that tetrodotoxin exists not only in pufferfish, but also in go...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/533G01N21/64
CPCG01N21/6486G01N33/533
Inventor 兰艺凤卫艳丽宋秀丽董川
Owner SHANXI UNIV