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Colloidal crystal gel label-free visual detection method with aptamer as identification unit

A technology of colloidal crystals and detection methods, which is applied in the direction of analyzing materials through chemical reactions, can solve the problems of high cost, poor selectivity of non-labeled visual detection, etc., and achieve high water content, modifiable detection objects, and excellent biological Effects of Compatibility and Stimuli Responsiveness

Inactive Publication Date: 2012-07-18
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the disadvantages of poor selectivity and high cost of existing non-labeled visual detection, the present invention provides a colloidal crystal gel non-labeled visual detection method using aptamers as molecular recognition units

Method used

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  • Colloidal crystal gel label-free visual detection method with aptamer as identification unit
  • Colloidal crystal gel label-free visual detection method with aptamer as identification unit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 The aptamer-modified colloidal crystal gel film detects heavy metal ions Hg in environmental water samples 2+

[0039] 1. Preparation of colloidal crystal hydrogel film

[0040] Add monodisperse silica colloidal nanoparticles with a diameter of 200 nanometers into the mixed solution of acrylamide and methylene bisacrylamide (29:1, molar ratio), adjust the mass percentage of silica to 30%, and ultrasonically disperse, Until the colloidal nanoparticle solution produces a bright and bright structural color; add photoinitiator 1173 (0.1%, mass volume ratio) to this solution, mix thoroughly, pass nitrogen for 5 minutes, and add 500 microliters of colloidal solution to the mold Inside, let it stand for 20 minutes and irradiate it with a high-pressure mercury lamp at 4°C for 15 minutes to obtain a colloidal crystal hydrogel film.

[0041] 2. Preparation of aptamer modified colloidal crystal hydrogel film

[0042] After reducing the colloidal crystal gel membrane prepared in...

Embodiment 2

[0045] Example 2 Using aptamer-modified colloidal crystal gel film to detect drug cocaine

[0046] 1. Preparation of colloidal crystal hydrogel film

[0047] Add monodisperse polystyrene colloidal nanoparticles with a diameter of 150 nanometers into the mixed solution of polyethylene glycol diacrylate and acrylamide (1:10, molar ratio), adjust the mass percentage of polystyrene to 40%, ultrasonic Disperse until the colloidal nanoparticle solution produces a bright and bright structural color; add photoinitiator 907 (0.1%, mass volume ratio) to this solution, mix thoroughly, pass nitrogen for 10 minutes, and take 500 microliters of colloidal solution Put it into a mold, let it stand for 10 minutes, and irradiate it with a high-pressure mercury lamp at 4°C for 20 minutes to obtain a colloidal crystal hydrogel film.

[0048] 2. Preparation of aptamer modified colloidal crystal hydrogel film

[0049] After reducing the colloidal crystal gel membrane prepared in the first step in 0.3M NaO...

Embodiment 3

[0052] Example 3 Detection of platelet-derived growth factor using aptamer-modified colloidal crystal gel film

[0053] 1. Preparation of colloidal crystal hydrogel film

[0054] Add monodisperse polymethyl methacrylate colloidal nanoparticles with a diameter of 100 nanometers into a 10% (mass to volume ratio) PEGDA solution, adjust the mass percentage of polymethyl methacrylate to 50%, and ultrasonically disperse until the colloidal solution Produce translucent and bright structural color; add initiator ammonium persulfate (0.1%, mass-volume ratio) and accelerator TEMED (0.1%, mass-volume ratio) to this solution, mix thoroughly, pass nitrogen for 30 minutes, take 500 A microliter volume of the colloidal solution was added to the mold, allowed to stand for 30 minutes, and polymerized in an oven at 40°C for 40 minutes to obtain a colloidal crystal hydrogel film.

[0055] 2. Preparation of aptamer modified colloidal crystal hydrogel film

[0056] After reducing the colloidal crystal ge...

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Abstract

The invention discloses a colloidal crystal gel label-free visual detection method with an aptamer as an identification unit. According to the method, a colloidal crystal hydrogel film is used as a carrier, the aptamer is combined with the colloidal crystal hydrogel film, and target molecules are detected by the high-specificity and high-sensitivity combination of the aptamer and a target detected object. After the aptamer is combined with the target molecules, the change of the conformation of the aptamer causes the change of the volume of a colloidal crystal gel, and the change of the volume of the gel is directly embodied by the change of a reflective peak of colloidal crystals (namely the color of the colloidal crystals), namely the existence or amount of a substance to be measured is embodied by the displacement of the reflective peak or the color of the colloidal crystal hydrogel film. The aptamer-based colloidal crystal hydrogel film label-free visual detection method has the advantages of high sensitivity and specificity, easiness for operation, low detection cost and the like.

Description

Technical field [0001] The invention relates to a non-marked visual detection method, in particular to a non-marked visual detection method using a colloidal crystal gel film with an aptamer as an identification unit as a carrier. Background technique [0002] The biosensor is a simple, fast and cheap important detection tool. Among the molecular recognition components of biosensors, antibodies are currently the most mature application. However, the application of antibodies is limited due to the cumbersome preparation, high cost, long time-consuming, harsh storage conditions, and easy inactivation of immobilization. Exploring new, highly specific and stable molecular recognition substances and establishing highly selective and sensitive biosensing technologies have become one of the frontier topics in modern analytical chemistry research. With the completion of the Human Genome Project and the development of nucleic acid research, especially when American scientists Tuerk and ...

Claims

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Application Information

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IPC IPC(8): G01N21/75
Inventor 顾忠泽叶宝芬赵远锦谢卓颖程瑶上官凤栖
Owner SOUTHEAST UNIV
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