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Bidirectional amplification ratio type electrochemical aptamer sensor and application thereof

An aptamer sensor, bidirectional amplification technology, applied in scientific instruments, biochemical equipment and methods, instruments, etc., can solve the problems of weak anti-interference and low sensitivity of alpha-fetoprotein, and achieve good anti-interference performance and wide detection range. Effect

Active Publication Date: 2020-04-14
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have weak anti-interference and low sensitivity for AFP detection

Method used

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  • Bidirectional amplification ratio type electrochemical aptamer sensor and application thereof
  • Bidirectional amplification ratio type electrochemical aptamer sensor and application thereof
  • Bidirectional amplification ratio type electrochemical aptamer sensor and application thereof

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Experimental program
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Effect test

Embodiment approach

[0034] A typical embodiment of the present invention provides a bidirectional amplified ratiometric electrochemical aptasensor, including capture electrodes, coded probes and RecJ f Exonuclease;

[0035] The capture electrodes include gold electrodes, DNA 1 and capture probes, the DNA 1 for single-stranded DNA, DNA 1 The 5' end of the capture probe is connected to the gold electrode, the capture probe is a hairpin structure protruding from the 3' end of the single-stranded DNA, the 3' of the capture probe is connected to ferrocene, and the 3' end of the capture probe is connected to the DNA 1 Complementary hybridization, the DNA sequence of the capture probe consists of sequence I and sequence II from the 3' end to the 5' end, and the sequence I is an aptamer for alpha-fetoprotein;

[0036] The coded probes include gold nanoparticles, DNA 2 and methylene blue, the DNA 2 For single-stranded DNA, the DNA 2 To be composed of sequence III and sequence IV from the 5' end to t...

Embodiment 1

[0055] Experimental reagents and materials:

[0056] Alpha-fetoprotein, carcinoembryonic antigen (CEA), human prostate-specific antigen (PSA), thrombin (TB) and human immunoglobulin (IgG) were purchased from Shanghai Lingchao Biotechnology Co., Ltd. (Shanghai, China). RecJ f Exonuclease and 10×NEBuffer were purchased from New England Biolabs (Beijing) Co., Ltd. (Beijing, China). Gold chloride, potassium ferricyanide and potassium ferrocyanide were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Tris-HCl (Tris-HCl) was purchased from Sangon Bioengineering (Shanghai) Co., Ltd. (Shanghai, China). 6-Mercapto-1-hexanol (MCH) and tris(2-carboxyethyl)phosphine hydrochloride (TCEP) were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Methylene blue was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). All other chemicals were of analytical reagent grade and used without further purificatio...

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Abstract

The invention discloses a bidirectional amplification ratio type electrochemical aptamer sensor and an application thereof. The bidirectional amplification ratio type electrochemical aptamer sensor comprises a capture electrode, a coding probe and a RecJf excision enzyme; the capture electrode comprises a gold electrode, DNA1 and a capture probe, the 5' end of the DNA1 is connected with the gold electrode, the capture probe is of a hairpin structure formed by single-stranded DNA and provided with a protruding 3' end, the 3' end of the capture probe is connected with ferrocene, the protruding part of the 3' end of the capture probe and DNA1 are hybridized and complemented, the DNA sequence of the capture probe is composed of a sequence I and a sequence II from the 3 'end to the 5' end, andthe sequence I is an aptamer of alpha fetoprotein; and the coding probe comprises gold nanoparticles, DNA2 and methylene blue, the DNA2 is composed of a sequence III and a sequence IV from the 5' endto the 3' end, the sequence IV is arranged to be hybridized and complementary with the DNA1, the 5' end of the DNA2 is connected with the gold nanoparticles, and the methylene blue is connected to theDNA2 through a G-quadruplex structure formed by the sequence IV.

Description

technical field [0001] The invention relates to a two-way amplified ratio electrochemical aptamer sensor and its application. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] In the early stage of the disease, reliable trace biomarker detection is of great significance for reducing the morbidity and mortality of the disease, avoiding long-term ineffective treatments and reducing side effects. Therefore, the development of methods for rapid and sensitive detection of biomarkers has attracted great attention in the fields of clinical diagnosis, biological research, and medical treatment. To date, various types of biosensors have been used for the detection of biomarkers. A...

Claims

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Application Information

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IPC IPC(8): C12Q1/682C12Q1/6825G01N27/327
CPCC12Q1/682C12Q1/6825G01N27/3278C12Q2565/519C12Q2525/301C12Q2525/205C12Q2563/137C12Q2521/319C12Q2565/607
Inventor 朱烨杨兴东李丹
Owner SHANDONG UNIV