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lncrna MEG3 dual detection electrochemical gene sensor, its preparation method and application

A dual detection, electrochemical technology, used in biochemical equipment and methods, scientific instruments, instruments, etc., to achieve high sensitivity, accelerated electron transfer, and high sensitivity

Active Publication Date: 2020-04-03
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no relevant report on the simultaneous detection of lncRNA two-component signature sequences by electrochemical biosensors

Method used

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  • lncrna MEG3 dual detection electrochemical gene sensor, its preparation method and application
  • lncrna MEG3 dual detection electrochemical gene sensor, its preparation method and application
  • lncrna MEG3 dual detection electrochemical gene sensor, its preparation method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0029] This embodiment illustrates the preparation method of the gene sensor of the present invention, such as figure 1 shown, including the following steps:

[0030] (1) Preparation of tungsten disulfide / dendritic gold nanocomposite modified electrodes by electrochemical deposition;

[0031] The sensor is a sensor constructed with a three-electrode system, the gold electrode is the working electrode, the saturated calomel electrode is the reference electrode, and the platinum electrode is the counter electrode.

[0032] The gold electrode was used as the working electrode, and the gold electrode was ultrasonically cleaned in acetone, ethanol, and water in sequence, and then dried with nitrogen gas. Take 20 μL of 1mg / mL tungsten disulfide dispersion and drop it on the surface of the gold electrode that has been cleaned, and dry it in the air to obtain a tungsten disulfide-modified gold electrode. Then use the electrochemical deposition method, set the deposition voltage -0.1V...

Embodiment 2

[0057] In this example, the mechanism verification of the gene sensor in Example 1 is carried out.

[0058] Set up 4 experimental groups, respectively without test object and with single test object T 1 , adding a single detection object T 2 and join the double detection object (T 1 and T 2 ). Using DPV for electrochemical detection, the results are as follows figure 2 shown.

[0059] It can be seen that curve a has no electrochemical signal response when no detection object is added. When adding a single detection object T 1 , the curve b has a single detection object T 1 Corresponding electrochemical response of ferrocene. When adding a single detection object T 2 , the curve c has a single detection object T 2 The corresponding electrochemical response of methylene blue. At the same time, add the detection object T 1 and T 2 When , the electrochemical signal response of ferrocene and methylene blue appears simultaneously in curve d. This shows that the electr...

Embodiment 3

[0061] In this embodiment, the detection results of the sensor are compared when the dual signal amplification strategy is adopted and when no signal amplification is adopted.

[0062] The preparation method of the sensor is as described in Example 1, and the present invention uses RNaseA auxiliary signal amplification and IT 1 and IT 2 The resulting hybridization chain reaction performs double signal amplification.

[0063] From image 3 It can be seen that the gene sensor prepared without dual signal amplification strategy in curve a has no obvious electrochemical response, while the gene sensor prepared with dual signal amplification strategy in curve b shows excellent electrochemical response peaks of ferrocene and methylene blue at the same time. This shows that the gene sensor of the present invention can effectively enhance the electrochemical response and ultrasensitively detect ultra-trace biomolecules.

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Abstract

The invention relates to an lncRNA MEG3 dual detecting electrochemical gene sensor, a preparation method and an application thereof. On the basis of an enzyme assisted target circulation signal amplification and DNA chain hybridizing (HCR) signal enhancement strategy, the ultra-sensitive quantitative analysis for double component is realized through a double signal probe, under the condition of adopting a tungsten disulfide / dendritic gold nanocomposite for modifying an electrode and adopting specific complementary base pairing. The linear scope of double component detected by the electrochemical gene sensor provided by the invention is 1fM-100pM; the detection limitation is low, respectively 0.25fM and 0.3fM; due to high sensitivity, repeatability, specificity, stability and low cost, thelncRNA MEG3 dual detecting electrochemical gene sensor has higher application value in the fields of biochemical research, clinical analysis, and the like.

Description

technical field [0001] The invention belongs to the technical field of material preparation and detection analysis, and in particular relates to an lncRNA MEG3 double detection electrochemical gene sensor, its preparation method and application. Background technique [0002] lncRNA MEG3 has broad prospects as a marker for early diagnosis and prognostic effect detection of lung cancer. However, ultrasensitive simultaneous detection of two lncRNA signature sequences remains challenging. Electrochemical sensors are often used in the construction of nucleic acid electrochemical sensors due to their excellent characteristics such as simplicity, high efficiency, low cost, and high sensitivity. At present, there is no related report on the simultaneous detection of lncRNA two-component signature sequences by electrochemical biosensors. Contents of the invention [0003] The object of the present invention is to provide a lncRNA MEG3 dual detection electrochemical gene sensor, i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327C12Q1/6825
CPCC12Q1/6825G01N27/3272G01N27/3276G01N27/3277G01N27/3278C12Q2563/143C12Q2563/149
Inventor 陈晓君李小燕黄和彭钢崔枫仇倩颖
Owner NANJING TECH UNIV
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