Method for detecting carcinoembryonic antigen by nano-probe-induced enzyme polymerization amplification electrochemical aptamer sensor

A carcinoembryonic antigen and nucleic acid aptamer technology, applied in the field of electrochemical biosensor research, can solve the problems of high cost, increase cost, increase the complexity of the reaction system, etc., and achieve strong applicability, high sensitivity, easy simplification and miniaturization. the effect of

Active Publication Date: 2017-03-01
NANJING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Polymerase chain reaction technology needs to strictly control the temperature change, so it needs relatively complex reaction equipment, and the cost is high; and the rolling circle replication amplification process also needs to add reaction templates, which increases the complexity of the reaction system and increases the cost.

Method used

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  • Method for detecting carcinoembryonic antigen by nano-probe-induced enzyme polymerization amplification electrochemical aptamer sensor
  • Method for detecting carcinoembryonic antigen by nano-probe-induced enzyme polymerization amplification electrochemical aptamer sensor
  • Method for detecting carcinoembryonic antigen by nano-probe-induced enzyme polymerization amplification electrochemical aptamer sensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: the preparation of nanoprobe

[0038] The method for detecting carcinoembryonic antigen by using the nanoprobe-induced enzyme polymerization amplification electrochemical nucleic acid aptamer sensor of the present invention first needs to prepare the nanoprobe. The nanoprobe preparation steps are as follows: centrifuge and concentrate 1000 mL of gold nanoparticle solution to 300 μL, and its final concentration is about 1 nmol / L; slowly add different volumes of 5′ terminal thiol-labeled carcinoembryonic antigen nucleic acid at a concentration of 100 μmol / L. Body signal probe solution; shake and react at 350rpm for 16 hours, add an equal volume of 0.02mol / L PBS (PH=7, 0.2mol / L NaCl), and age for 40 hours; centrifuge at 10000rpm for 15 minutes, add 0.01mol / L PBS (PH=7, 0.1mol / L NaCl), centrifuged at 10000rpm for 15 minutes, and resuspended in PBS (0.01M, 0.25M NaCl) solution containing 1wt% Tween to prepare a nanoprobe solution. Ultraviolet detection was ca...

Embodiment 2

[0039] Example 2: The effect of the signal probe concentration of the 5'-end sulfhydryl-labeled carcinoembryonic antigen nucleic acid aptamer on the detection results of electrochemical signals.

[0040] Using the nanoprobe-induced enzyme polymerization amplification electrochemical nucleic acid aptamer sensor detection method for carcinoembryonic antigen of the present invention, using the carcinoembryonic antigen solution as the target, building a nanoprobe-induced enzyme polymerization amplification electrochemical nucleic acid aptamer sensor system , all the operation steps are as above, wherein the concentration of the assembled 3'-end sulfhydryl-labeled carcinoembryonic antigen nucleic acid aptamer capture probe is 3 μmol / L, the concentration of blocking solution OEG is 1 mmol / L, and the concentration of carcinoembryonic antigen is 1 μg / mL , the nanoprobe adopts the nanoprobe preparation step described in Example 1, wherein the final concentration of the added 5' end sulf...

Embodiment 3

[0041] Example 3: The influence of the assembly density of the 3'-end sulfhydryl-labeled carcinoembryonic antigen nucleic acid aptamer capture probe on the detection results of electrochemical signals.

[0042] The method for detecting carcinoembryonic antigen by using nanoprobe-induced enzyme polymerization and amplifying electrochemical nucleic acid aptamer sensor of the present invention takes carcinoembryonic antigen solution as the target object, and all the operation steps are as described in Example 2, wherein the concentration of the blocking solution OEG It is 1mmol / L, and the concentration of carcinoembryonic antigen is 1 μ g / mL, and the nanoprobe adopts the nanoprobe preparation step described in embodiment 1, wherein the added 5' end sulfhydryl label carcinoembryonic antigen nucleic acid aptamer signal probe The final concentration was 3 μmol / L, and the concentration of the assembled 3'-end sulfhydryl-labeled carcinoembryonic antigen nucleic acid aptamer capture pro...

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Abstract

The invention discloses a method for detecting a carcinoembryonic antigen by a nano-probe-induced enzyme polymerization amplification electrochemical aptamer sensor. The method comprises that the surface of a gold nanoparticle are modified through an end 5' mercapto-labeled carcinoembryonic antigen aptamer signal probe so that a nanoprobe is prepared, a layer of end 3' mercapto-labeled carcinoembryonic antigen aptamer capture probes modifies the surface of a gold electrode, after addition of a carcinoembryonic antigen and the nanoprobes, the 3' mercapto-labeled carcinoembryonic antigen aptamer capture probe, the carcinoembryonic antigen and the end 5' mercapto-labeled carcinoembryonic antigen aptamer signal probe are specifically bonded so that the exposed end 3' is formed on the electrode surface and the tail end is extended, then an avidin-labeled horseradish peroxidase is specifically bonded to the electrode surface, and change of an electrochemical signal produced by a horseradish peroxidase catalytic electrolyte is detected so that high sensitivity and high specificity detection of a carcinoembryonic antigen is realized. The method is used for early diagnosis of tumors, curative effect judgment, disease progression, monitoring and prognosis estimation.

Description

technical field [0001] The invention belongs to the research field of electrochemical biosensors, and relates to a method for detecting carcinoembryonic antigens by nanometer probes inducing enzyme polymerization and amplifying electrochemical nucleic acid aptamer sensors. Background technique [0002] On the one hand, carcinoembryonic antigen is a broad-spectrum tumor marker, which plays an important role in the judgment of curative effect, disease development, monitoring and prognosis estimation of various tumors; on the other hand, the content of carcinoembryonic antigen in actual samples is often Very low, it is necessary to research and design a highly sensitive and highly selective detection method to realize the detection of carcinoembryonic antigen. In order to improve the detection sensitivity and selectivity of carcinoembryonic antigen, various carcinoembryonic antigen detection methods have emerged as the times require. It has become the focus of research to reali...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N33/68
Inventor 万莹王鹏娟苏岩杨树林
Owner NANJING UNIV OF SCI & TECH
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