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Evanescent wave aptamer sensor and method for detecting small molecules by using evanescent wave aptamer sensor

An aptamer sensor and nucleic acid aptamer technology, applied in the field of analytical chemistry, can solve the problems of limited specificity and low sensitivity, and achieve the effects of high specificity, high detection sensitivity and low test cost

Active Publication Date: 2020-12-15
深圳市翠云谷科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, many nucleic acid aptamer sensors have been reported for the above three targets, including electrochemical and optical sensors, but they all have problems of low sensitivity or limited specificity and have not been used in practical applications.

Method used

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  • Evanescent wave aptamer sensor and method for detecting small molecules by using evanescent wave aptamer sensor
  • Evanescent wave aptamer sensor and method for detecting small molecules by using evanescent wave aptamer sensor
  • Evanescent wave aptamer sensor and method for detecting small molecules by using evanescent wave aptamer sensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. The principle of nucleic acid aptamer evanescent wave fiber sensor (SEP-AS / EWA) with target self-enrichment and purification capabilities, optical fiber preparation, target testing and regeneration process of its sensing interface.

[0035] The invention provides a nucleic acid aptamer evanescent wave optical fiber biosensor (SEP-AS / EWA) with target self-enrichment and purification capabilities and a method for using it to realize high sensitivity and high specificity detection of small molecule targets. The principle of the inventive method is as Figure 1A and 1B As shown, the extraction agent (such as Tween 80) with high-efficiency target extraction ability and the target-specific nucleic acid aptamer were assembled on the optical fiber sensing interface to realize the simultaneous progress of target enrichment, purification and specific detection. The sensitivity and specificity are very high. The method of the present invention realizes the quantitative...

Embodiment 2

[0042] Example 2. Effect of interface chemistry on EWA for DEHP detection.

[0043] See Table 1 for the sequence information of the DNA probes used in the following examples. DEHP standard solution configuration and detection both use buffer solution 1 (100mM sodium chloride, 20mM tris, 2mM magnesium chloride, 5mM potassium chloride, 1mM calcium chloride, 1% Tween 20, 0.03% triton X-100, 2% dimethyl sulfoxide, pH 7.9); SDM standard solution configuration and detection all use buffer solution 2 (25mM 4-hydroxyethylpiperazineethanesulfonic acid, 20mM potassium chloride, 200mM sodium nitrate, 150mM ammonium chloride, 0.025% Triton X-100, 1% dimethyl sulfoxide, pH 5.3); Kana standard solution configuration and detection use buffer solution 3 (10mM phosphate buffer solution, 1M sodium chloride, 5 mM MgCl2, pH 7.0).

[0044] According to the method described in Example 1, three evanescent wave sensors with different interface chemistries were respectively constructed for the detec...

Embodiment 3

[0046] Example 3. High sensitivity and high specificity detection of DEHP using SEP-AS / EWA.

[0047]The steps of hydroxylation, silanization, coupling, blocking and reduction of the optical fiber surface are the same as in Example 1, wherein the nucleic acid aptamer coupled is NH2-PAE. DEHP standard solutions (0pM, 1pM, 10pM, 100pM, 1nM, 10nM, 100nM) with different final concentrations were prepared. Mix with 100nM fluorescence-modified complementary chain (c-PAE-Cy5.5) respectively, pass it into the evanescent wave fiber optic sensor from low concentration to high concentration in sequence, regenerate the interface after each test is completed, clean the pipeline, and make it Fluorescent signal drops to baseline. The change of fluorescence with time at different concentrations was recorded, and the working curve was drawn on the ordinate with the relative fluorescence signal decrease percentage value at different target concentrations.

[0048] Configure different final con...

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Abstract

The invention relates to an evanescent wave nucleic acid aptamer sensor and a method for detecting small molecules by using the evanescent wave nucleic acid aptamer sensor, and belongs to the technical field of analytical chemistry. According to the invention, the extraction agent with efficient target extraction capability and the target specific aptamer are assembled on the optical fiber sensinginterface together, so that target enrichment, purification and specific detection are carried out synchronously, and the detection sensitivity and specificity are extremely high. Based on competitive combination of the small molecular target, the short-chain DNA complementary to the aptamer and the aptamer coupled to the surface of the optical fiber, quantitative detection of the small moleculartarget is realized. The detection limit of the method is many times lower than that of a traditional EWA detection method, and the method has excellent specificity. Detection is very rapid, and regeneration of the sensor can be rapidly achieved. The sensor and the detection method have practical application value and universality, and can be popularized and applied to detection of other types oftargets such as metal ions and proteins by utilizing other extraction reagents.

Description

technical field [0001] The present invention relates to an evanescent wave nucleic acid aptamer sensor and a method for detecting small molecules using the same, in particular to an evanescent wave nucleic acid aptamer sensor (SEP-AS / EWA) capable of target self-enrichment and purification. ), and using the sensor to realize highly sensitive and highly specific detection of various water-soluble small molecule targets, which belongs to the technical field of analytical chemistry. Background technique [0002] Small organic molecules are a large category of environmental and food pollutants. They have the characteristics of various types, large differences in water solubility, and few specific antibodies. Therefore, methods based on large instruments are mainly used in analysis and testing, such as gas chromatography. , high performance liquid chromatography (HPLC), gas chromatography mass spectrometry, liquid chromatography mass spectrometry, etc. These methods require expen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533
CPCG01N33/533C12N2310/3517C12N2310/16C12N15/115G01N33/5308G01N33/54373G01N21/648G01N2021/6484
Inventor 娄新徽陆张伟
Owner 深圳市翠云谷科技有限公司