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Construction method for electrochemical sensor used for DNA single chain and protein molecule detection

A molecular detection and construction method technology, applied in the field of analysis and detection, can solve the problems of low signal value, low sensitivity, and high detection limit concentration of electrochemical sensors, and achieve high sensitivity, improve detection sensitivity, and low detection limit concentration.

Active Publication Date: 2013-01-16
珠海中科先进科技产业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Based on this, the present invention provides an electrochemical sensor that can be used for the detection of DNA single strands and specific protein molecules to solve the shortcomings of traditional electrochemical sensors with low signal value, low sensitivity and high detection limit concentration

Method used

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  • Construction method for electrochemical sensor used for DNA single chain and protein molecule detection
  • Construction method for electrochemical sensor used for DNA single chain and protein molecule detection
  • Construction method for electrochemical sensor used for DNA single chain and protein molecule detection

Examples

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Effect test

Embodiment 1

[0030] Such as figure 1 As shown, a gold disk electrode 110 with a diameter of 2.0 mm is taken, and then polished with aluminum oxide powder with a particle size of 1.0 mm, 0.3 mm, and 0.05 mm respectively, and then the gold disk electrode 110 is placed in two successively. Ultrasonic cleaning in sub-water, ethanol and acetone for many times. The cleaned gold disk electrode 110 is successively put into 0.5M sodium hydroxide solution and 0.5M sulfuric acid solution for electrochemical cycle scanning, washed with secondary water for several times, and then the gold disk electrode 110 is blown dry with high purity nitrogen. The treated gold disc electrode 110 was mixed with 0.5 μM capture probe 120 and 5.0 μM tris(2-carbonyl) in a 200 mM tris-hydrochloric acid buffer solution (hereinafter referred to as Tris-HCl) at pH 7.4. Ethyl)phosphorus hydrochloride was reacted at a temperature of 37°C and a duration of 16 hours. The DNA sequence of the capture probe 120 was: 5′-HS-(CH 2 ) 6 ...

Embodiment 2

[0034] Such as Figure 4 As shown, the pretreatment process of the gold disk electrode 210 is the same as in the first embodiment. The treated gold disk electrode 210 reacts with 0.5μM capture probe 220 and 5.0μM tris(2-carbonylethyl)phosphorus hydrochloride in a 200mM Tris-HCl buffer solution with a pH of 7.4 and a concentration of 200mM. The reaction temperature is 37℃, the duration is 16 hours, the capture probe 220 is the aptamer sequence of thrombin: 5′-HS-(CH 2 ) 6 -AGACAA GGA AAA TCC TTC CCC CCC CGG TTG GTG TGG TTG G-3'. Under the same buffer solution and reaction conditions, the gold disk electrode 210 is further reacted with 0.5μM additional probe 230. The DNA sequence of the additional probe 230 is: 5′-GGT TGG TGT GGT TGG-(CH 2 ) 3 -SH-3'. After the gold disk electrode 210 modified by the capture probe 220 and the additional probe 230 is washed with Tris-HCl buffer solution, it is further reacted with 6-mercaptoethanol in a 10 mM Tris-HCl buffer solution of pH 7.4 at ...

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Abstract

The invention relates to a construction method for an electrochemical sensor used for DNA single chain and protein molecule detection. The construction method achieves a signal enhanced effect of DNA single chain detection by introducing a unique additional DNA probe on the basis of a design of a conventional DNA electrochemical sensor, and can be further applied to the protein electrochemical sensor based on a nucleic acid aptamer. The construction method overcomes the intrinsic disadvantages of poor detection sensitivity, tedious pretreatment processes and high concentration for detection limit in a conventional electrochemical sensor used for the DNA single chain and protein molecule detection, and has very good popularized value in the field of the DNA electrochemical sensor.

Description

【Technical Field】 [0001] The invention relates to analysis and detection technology, in particular to an electrochemical sensor for DNA single-strand and protein molecule detection. 【Background technique】 [0002] In recent years, due to the needs of rapid development in the fields of early diagnosis of diseases, rapid and accurate detection of biomarkers, and environmental monitoring, more and more biosensor devices that can be used for molecular detection of DNA and specific proteins have attracted attention. Among them, electrochemical sensors have gradually become the focus of research and development due to their low production cost, rapid detection, and portability. They are also considered to be one of the most valuable DNA and specific protein molecules with high sensitivity detection methods. In the research and development process of electrochemical sensors, the improvement of detection sensitivity is the most critical issue. However, the traditional electrochemical se...

Claims

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Application Information

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IPC IPC(8): G01N27/26G01N27/30
Inventor 张春阳杨坤
Owner 珠海中科先进科技产业有限公司
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