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Preparation method and application of electrochemical biosensor for detecting NSCLC circulating tumor gene

A biosensor and tumor gene technology, applied in the field of electrochemical DNA sensors, can solve the problems of electrochemical biosensors that have not been reported yet, and achieve the effects of improving water solubility, avoiding aggregation, and low price

Pending Publication Date: 2021-04-30
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, no electrochemical biosensors have been reported for the detection of EGFR L858R in blood.

Method used

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  • Preparation method and application of electrochemical biosensor for detecting NSCLC circulating tumor gene
  • Preparation method and application of electrochemical biosensor for detecting NSCLC circulating tumor gene
  • Preparation method and application of electrochemical biosensor for detecting NSCLC circulating tumor gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] The construction of embodiment 1 electrochemical DNA sensor

[0060] Follow the steps below (construct the schematic diagram as figure 1 shown):

[0061] Step 1: Dissolve 1 mL of 1% HAuCl 4 Add the solution to 100mL ultrapure water and boil, then quickly add 2.5mL 1% trisodium citrate solution and continue to boil for 15 minutes. After cooling, restore the original volume to the original volume with ultrapure water to obtain a transparent wine-red solution that is gold nanoparticles (AuNPs). For the construction principle of step 1, see figure 1 Middle (A) panel.

[0062] Step 2: Dissolve 0.461mmol 2-aminoterephthalic acid in 10mL N,N-dimethylformamide, then dissolve 0.461mmol FeCl 3 ·6H 2 O was added thereto, and 2.3 mmol acetic acid was added to the mixture, cooled to room temperature in an oil bath at 120°C for 4 h, centrifuged, washed with DMF and absolute ethanol three times, and the resulting precipitate was vacuum-dried at 60°C, and finally the brown powder ...

Embodiment 2

[0077] Example 2 Detection of NSCLC circulating tumor gene (EGFR L858R) by electrochemical DNA sensor

[0078] The electrochemical DNA sensor constructed in Example 1 was used to detect the NSCLC circulating tumor gene EGFR L858R, and the following steps were followed:

[0079] (1) Drawing of standard curve:

[0080] The DNA sensor constructed in Example 1 was placed in a solution containing 1.7mM K 4 Fe(CN) 6 and 0.24M KCl in 0.1M PBS solution to measure the current value of different concentrations of ctDNA. The standard curve was drawn according to the logarithmic value of different concentrations of ctDNA and the DPV response signal. The test results showed that the two had a good linear relationship in the concentration range of 100fM-100nM, the linear correlation coefficient was 0.9972, and the detection limit was 25.7fM. For the results, see figure 2 . figure 2 In (A) is containing 1.7mM K 4 Fe(CN) 6 and 0.24mM KCl in 0.1M PBS buffer, the differential pulse volt...

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Abstract

The invention discloses a preparation method and application of an electrochemical biosensor for detecting an NSCLC circulating tumor gene. The preparation method comprises the following steps of taking a prepared NG-PEI-COF nano composite material as a substrate material of the sensor, and taking Prussian blue generated by Fe-MOF and Fe<2+> as a signal amplification material; the sandwich type biosensor for detecting the circulating tumor gene EGFR L858R is constructed by utilizing nucleic acid hybridization. The method is advantaged in that the biosensor provided by the invention not only has a stronger signal amplification effect, but also has advantages of wide linear range, strong specificity, short analysis time and the like, and can provide a new method for diagnosing the non-small cell lung cancer.

Description

technical field [0001] The invention relates to the technical field of electrochemical DNA sensors, in particular to a method for preparing an electrochemical biosensor for detecting NSCLC circulating tumor gene EGFR L858R, and the application of the electrochemical biosensor in detecting serum EGFRL858R. Background technique [0002] Lung cancer is one of the most common malignancies in the world and has become the leading cause of cancer-related death worldwide. Lung cancer is divided into small cell lung cancer and non-small cell lung cancer. Among them, non-small cell lung cancer (NSCLC) accounts for about 85% of all lung cancers, and about 75% of NSCLC patients are already at an advanced stage when they are discovered, and their 5-year survival rate is very high. Low. Existing diagnostic methods for NSCLC have limitations to a certain extent, such as high price, complicated and time-consuming process, and great damage to the patient's body. Therefore, for non-small ce...

Claims

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Application Information

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IPC IPC(8): G01N27/26G01N27/30G01N27/327
CPCG01N27/26G01N27/30G01N27/3278
Inventor 白丽娟母昭德郭蕾
Owner CHONGQING MEDICAL UNIVERSITY
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