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Sensor based on hybridization chain reaction and ribozyme and carcino-embryonic antigen detection method

A hybrid chain reaction, carcinoembryonic antigen technology, applied in the field of chemical and biological sensing, can solve the problems of harsh experimental conditions, high cost, low sensitivity, etc., to improve selectivity and sensitivity, reduce detection costs, and shorten detection time. Effect

Active Publication Date: 2020-05-19
SHANXI DATONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Carcinoembryonic antigen is a broad-spectrum tumor marker, which has important clinical value for the differential diagnosis of malignant tumors, disease monitoring, and efficacy evaluation. However, in actual samples, the concentration of carcinoembryonic antigen is too low to achieve high sensitivity , highly selective detection
[0003] At present, the commonly used detection methods for carcinoembryonic antigen mainly include: electrochemical immunoassay, chemiluminescence immunoassay, colorimetric immunoassay, etc., but these methods have problems such as high cost, time-consuming, poor selectivity, low sensitivity, and harsh experimental conditions. These methods are restricted in practical application

Method used

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  • Sensor based on hybridization chain reaction and ribozyme and carcino-embryonic antigen detection method
  • Sensor based on hybridization chain reaction and ribozyme and carcino-embryonic antigen detection method
  • Sensor based on hybridization chain reaction and ribozyme and carcino-embryonic antigen detection method

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Experimental program
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Effect test

Embodiment 1

[0055] The nucleic acid aptamer probe, the first hairpin probe, the second hairpin probe and carcinoembryonic antigen were centrifuged at 10,000rpm and 30s at 4°C before opening the cover for the first time, and then dissolved in secondary water to make the concentration 100 μM mother solution, and then diluted with Tris-HCl buffer to form a nucleic acid aptamer probe solution, a first hairpin probe solution and a second hairpin probe solution, and stored at 4°C until use; the nucleic acid aptamer probe The needle solution 5 μM, the first hairpin probe solution 5 μM and the second hairpin probe solution 5 μM were respectively heated to 90° C., reacted for 10 minutes, and cooled to room temperature for use.

[0056] Among them, in figure 2 In (a), add Tris-HCl buffer solution with a pH of 7.4 to the centrifuge tube, and react at 37°C for 6 hours; 2 HPO 4 -NaOH buffer, react at room temperature for 20 minutes.

[0057] exist figure 2 In (b), Tris-HCl buffer solution with a...

Embodiment 2

[0063] The nucleic acid aptamer probe, the first hairpin probe, the second hairpin probe and the carcinoembryonic antigen are centrifuged at 10,000 rpm and 4°C for 30 seconds before opening the cover for the first time, and then dissolved in secondary water to make 100 μM Dilute the mother solution with Tris-HCl buffer and store at 4°C until use; heat the aptamer probe solution 5 μM, the first hairpin probe solution 5 μM and the second hairpin probe solution 5 μM to 90°C respectively , reacted for 10 minutes, and cooled to room temperature for later use.

[0064] Among them, in image 3 In (a), add 50nM nucleic acid aptamer probe solution, 50nM first hairpin probe solution and 50nM second hairpin probe solution to the centrifuge tube in Tris-HCl buffer solution with pH 7.4, and react at 37°C 6 hours to obtain a mixed solution; then add 0.5 μM hemin, react at room temperature for 2 hours to obtain a reaction solution; then take out 100 μL of the reaction solution and add 6 mM ...

Embodiment 3

[0075] The nucleic acid aptamer probe, the first hairpin probe, the second hairpin probe and carcinoembryonic antigen were centrifuged at 10,000rpm and 4°C for 30s before being used for the first time, and were dissolved in secondary water to prepare 100μM The mother solution was diluted with Tris-HCl buffer solution and stored at 4°C until use; the aptamer probe solution 5 μM, the first hairpin probe solution 5 μM and the second hairpin probe solution 5 μM were heated to 90°C, React for 10 minutes, and cool to room temperature for later use.

[0076] exist Figure 5 In (a), add 50nM nucleic acid aptamer probe solution, 50nM first hairpin probe solution, 50nM second hairpin probe solution and 0.25nM immunoglobulin G (IgG) to the centrifuge tube at pH 7.4 In Tris-HCl buffer solution, react at 37°C for 6 hours to obtain a mixed solution; then add 0.5 μM hemin, react at room temperature for 2 hours to obtain a reaction solution; then take out 100 μL of the reaction solution and ...

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Abstract

The invention relates to the technical field of chemical and biological sensing, and particularly a sensor based on hybridization chain reaction and ribozyme and a carcino-embryonic antigen detectionmethod. The sensor based on the hybridization chain reaction and ribozyme comprises a nucleic acid aptamer probe and a hairpin probe, wherein the aptamer probe comprises an aptamer sequence, an initiation chain sequence and a complementary sequence, the hairpin probes comprise a first hairpin probe and a second hairpin probe, the first hairpin probe and the second hairpin probe comprise G-quadruplex ribozyme base sequences, the nucleic acid aptamer probe specifically recognizes a carcino-embryonic antigen, the aptamer probe is subjected to conformational transformation to make the first hairpin probe and the second hairpin probe form double-stranded DNA, moreover, in the presence of hemin, the first hairpin probe and the second hairpin probe are self-assembled to form hemin / G-quadruplex ribozyme, and the hemin / G-quadruplex ribozyme catalyzes and oxidizes thiamine mediated by hydrogen peroxide and emits fluorescence, so the carcino-embryonic antigen can be quantitatively detected.

Description

technical field [0001] The invention relates to the technical field of chemical and biological sensing, in particular to a sensor based on hybridization chain reaction and ribozyme and a carcinoembryonic antigen detection method. Background technique [0002] Carcinoembryonic antigen is a broad-spectrum tumor marker, which has important clinical value for the differential diagnosis of malignant tumors, disease monitoring, and efficacy evaluation. However, in actual samples, the concentration of carcinoembryonic antigen is too low to achieve high sensitivity , Highly selective detection. [0003] At present, the commonly used detection methods for carcinoembryonic antigen mainly include: electrochemical immunoassay, chemiluminescence immunoassay, colorimetric immunoassay, etc., but these methods have problems such as high cost, time-consuming, poor selectivity, low sensitivity, and harsh experimental conditions. These methods are restricted in practical application. Conten...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574C12Q1/6825C12Q1/682
CPCG01N33/57473C12Q1/6825C12Q1/682C12Q2525/205C12Q2525/301C12Q2521/337C12Q2563/107Y02A50/30
Inventor 白云峰张慧琳赵璐李婉宁冯锋王玉珍陈晓亮陈泽忠
Owner SHANXI DATONG UNIV
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