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Method for detecting carcino-embryonic antigens by using electrochemical nucleic acid aptamer sensor on basis of terminal elongases

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

Active Publication Date: 2017-03-22
迈科若(苏州)医疗科技有限公司
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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 carcino-embryonic antigens by using electrochemical nucleic acid aptamer sensor on basis of terminal elongases
  • Method for detecting carcino-embryonic antigens by using electrochemical nucleic acid aptamer sensor on basis of terminal elongases
  • Method for detecting carcino-embryonic antigens by using electrochemical nucleic acid aptamer sensor on basis of terminal elongases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: 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.

[0037] The method for detecting carcinoembryonic antigen by using the electrochemical nucleic acid aptamer sensor based on terminal elongase according to the present invention, taking the carcinoembryonic antigen solution as the target object, and the establishment steps of the carcinoembryonic antigen detection system based on the terminal elongase electrochemical sensor As mentioned above, the concentration of blocking solution OEG is 1 mmol / L, the concentration of carcinoembryonic antigen is 1 μg / mL, the concentration of carcinoembryonic antigen nucleic acid aptamer signal probe is 1 μmol / L, and the assembled 3' terminal thiol-labeled carcinoembryonic The concentration of the antigen nucleic acid aptamer capture probe was 0.2, 0.5, 1, 2 and 5 μmol / L, and the influence of dif...

Embodiment 2

[0038] Example 2: Effect of OEG blocking concentration on electrochemical detection results.

[0039] The method for detecting carcinoembryonic antigen using the terminal elongase-based electrochemical nucleic acid aptamer sensor of the present invention takes the carcinoembryonic antigen solution as the target object, and all the operating steps are as described in Example 1, wherein the assembled 3' terminal thiol The concentration of labeled carcinoembryonic antigen nucleic acid aptamer capture probe is 1 μmol / L, the concentration of carcinoembryonic antigen is 1 μg / mL, the concentration of carcinoembryonic antigen nucleic acid aptamer signal probe is 1 μmol / L, and the concentration of blocking solution OEG is 0.5, 1, 1.5 and 2mmol / L, analyze the impact of different blocking concentrations on the detection of electrochemical signals, the analysis results are as attached image 3 As shown, the OEG concentration is lower than 1mmol / L, and the signal-to-noise ratio is enhanced...

Embodiment 3

[0040] Example 3: Effect of concentration of carcinoembryonic antigen nucleic acid aptamer signal probe on electrochemical signal detection results.

[0041] The method for detecting carcinoembryonic antigen using the terminal elongase-based electrochemical nucleic acid aptamer sensor of the present invention takes the carcinoembryonic antigen solution as the target object, and all the operating steps are as described in Example 1, wherein the assembled 3' terminal thiol The concentration of labeled carcinoembryonic antigen aptamer capture probe was 1 μmol / L, the concentration of blocking solution OEG was 1 mmol / L, the concentration of carcinoembryonic antigen was 1 μg / mL, and the concentration of carcinoembryonic antigen aptamer signal probe was 0.2 , 0.5, 1, 2, and 5 μmol / L to analyze the influence of different carcinoembryonic antigen nucleic acid aptamer signal probe concentrations on the detection of electrochemical signals, and the analysis results are shown in the attach...

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PUM

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Abstract

The invention discloses a method for detecting carcino-embryonic antigens by using an electrochemical nucleic acid aptamer sensor on the basis of terminal elongases. The method comprises the following steps: modifying a layer of 3' mercapto-terminated labeled carcino-embryonic antigen aptamer capturing probes on the surface of a gold electrode at first; then adding carcino-embryonic antigens and carcino-embryonic antigen aptamer signal probes, wherein the 3' mercapto-terminated labeled carcino-embryonic antigen aptamer capturing probes and the carcino-embryonic antigen aptamer signal probes can specifically recognize the carcino-embryonic antigens, so that a sandwich structure is formed on the surface of the electrode; extending biotin labeled nucleic acid long chains at 3' terminals of the carcino-embryonic antigen aptamer signal probes under the effect of terminal deoxynucleotidyl transferase, wherein avidin labeled horse radish peroxidase can be combined to the biotin labeled nucleic acid long chains in a compatible manner; and detecting electrochemical signal change generated by horse radish peroxidase catalyzed electrolyte so as to realize high-sensitivity and high-specificity detection on the carcino-embryonic antigens. The method can be used for early diagnosis of tumors, curative effect judgment, progression of the disease, prognosis estimation and the like.

Description

technical field [0001] The invention belongs to the research field of electrochemical biosensors, and relates to a method for detecting carcinoembryonic antigens by an electrochemical nucleic acid aptamer sensor based on terminal elongation enzyme. Background technique [0002] 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; however, the content of carcinoembryonic antigen in actual samples is often very low, which needs to be studied And design a detection method with high sensitivity and high selectivity 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 realize the specific recognition and signal amplificat...

Claims

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Application Information

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IPC IPC(8): G01N27/26
Inventor 万莹王鹏娟苏岩杨树林
Owner 迈科若(苏州)医疗科技有限公司
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