Temperature controllable electrochemical mercury ion sensor and preparation method thereof

An electrochemical and sensor technology, applied in the field of nucleic acid detection and biological analysis, can solve the problems of increasing the electrode response signal, denaturing the molecular structure of the enzyme, shortening the sensor time, etc., to increase the electrode response signal, enhance the activity, and improve the detection sensitivity Effect

Active Publication Date: 2017-10-03
FUZHOU UNIVERSITY
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Problems solved by technology

[0004] Exonuclease III is a biological macromolecule that is very sensitive to temperature changes. When the temperature is lower than the optimum temperature, the activity of the enzyme is poor, but if the temperature is too high, the molecular structure of the enzyme will be irreversibly denatured and the enzyme will be inactivated.
Most of the amperometric biosensors reported in the past control the overall temperature change of the experimental

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  • Temperature controllable electrochemical mercury ion sensor and preparation method thereof
  • Temperature controllable electrochemical mercury ion sensor and preparation method thereof
  • Temperature controllable electrochemical mercury ion sensor and preparation method thereof

Examples

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Example Embodiment

[0037] Example 1

[0038] A temperature-controllable electrochemical Hg based on exonuclease III target cycle signal amplification 2+ The preparation method of the sensor, such as figure 1 As shown, including the following steps:

[0039] (1) Design a signal probe P1 labeled with ferrocene (Fc) close to the 5'. The complementary existence of the signal probe P1 and the DNA auxiliary probe P2 can recognize Hg 2+ The T-T mismatch structure in Hg 2+ In the presence, P2 and P1 hybridize to form a T-Hg 2 + -T structure of double-stranded structure with 3'flat ends. Induces exonuclease III to digest P1, Hg 2+ It is released for recycling; the 5'end of the P1 chain is thiolated;

[0040] Among them, the nucleotide sequence of the signal probe P1 is: 5’-SH-(CH 2 ) 6 -CCCCA T(Fc)CGCC ACCAG CTTCT -3’,

[0041] The nucleotide sequence of the auxiliary probe P2 is: 5’-TGTAG CTGGT GGCGA TCCCA C-3’;

[0042] (2) The gold plate hot electrode uses 0.05 mm Al on the suede 2 O 3 Polished into a mirror...

Example Embodiment

[0045] Example 2

[0046] Reaction time optimization experiment:

[0047] The cycle reaction time of restriction enzyme digestion in step (4) of Example 1 is 0, 15, 30, 45, 60, 75, 90, 105, 120 min. At different electrode temperatures (0 ºC, 25 ºC, 40 ºC) The experiment was carried out, and the other reaction conditions were the same as in Example 1. The sulfhydrylation signal probe P1 modified gold disk hot electrode obtained in step (3) of Example 1 was subjected to SWV detection in 10 mM tris-HCl detection solution to obtain Fc Oxidation peak current I 0(Fc) ; The electrochemical biosensor finally obtained in step (4) of Example 1 was detected with SWV in a 10 mM tris-HCl detection solution, and the obtained Fc oxidation peak current I Fc And get the I 0(Fc) The difference is definitely worth:|ΔI Fc |=|I Fc -I 0(Fc) |; with|ΔI Fc |Plot the temperature, such as figure 2 As shown, at the same electrode temperature, it can be seen that the duration of the reaction time |ΔI Fc...

Example Embodiment

[0048] Example 3

[0049] Reaction temperature optimization experiment:

[0050] Only change the reaction temperature in step (4) of Example 1, where a is the control group, and the reaction temperature of bf is 0, 10, 20, 24, 30, 35, and 40 ºC. Experiments are performed separately, and other reaction conditions are implemented simultaneously Example 1: The signal probe P1 modified gold disk hot electrode obtained in step (3) of Example 1 was subjected to SWV detection in 10 mM tris-HCl detection solution to obtain the oxidation peak current I of Fc 0(Fc) ; The electrochemical biosensor finally obtained in step (4) of Example 1 was detected with SWV in 10 mM tris-HCl detection solution, and the obtained Fc oxidation peak current was compared with the obtained I 0(Fc) The difference is definitely worth:|ΔI Fc |=|I Fc -I 0(Fc) |; with|ΔI Fc |Plot the temperature, such as image 3 As shown, it can be seen that as the temperature increases |ΔI Fc | Becomes larger, indicating that th...

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Abstract

The present invention discloses a temperature-controllable electrochemical Hg<2+> sensor based on exonuclease (Exo III) target circulating signal amplification, and a preparation method thereof. The sensor comprises a gold plate thermal electrode, a signal probe P1, an auxiliary probe P2 and exonuclease III, the T-T mismatching structure capable of identifying Hg<2+> can be produced through the complementation of P2 and P1, the T-T mismatching structure and Hg<2+> for a double-stranded structure having a T-Hg<2+>-T structure and having a 3' blunt end, the exonuclease III is induced to digest the P1, the amount of the P1 on the surface of the electrode is decreased so as to decrease the Fc chemical signal, and the decreasing degree of the Fc chemical signal on the electrode surface and the Hg<2+> concentration form linear relationship, such that the high sensitivity detection on the Hg<2+> can be achieved.

Description

technical field [0001] The invention belongs to the technical field of biological analysis, in particular to a temperature-controllable electrochemical mercury ion sensor and a preparation method thereof, which are applied in the field of nucleic acid detection. Background technique [0002] heavy metal ion Hg 2+ Due to their relatively stable properties, they are difficult to be biodegraded in the natural environment and have strong biological toxicity. They will eventually remain in the human body through the enrichment of the food chain and destroy various physiological functions of the human body. Human tissues and organs even pose a serious threat to life. Therefore, the establishment of a rapid and reliable detection method for mercury ions is of great significance in environmental monitoring and food safety testing. [0003] Traditional heavy metal ion detection methods mainly rely on large-scale professional instruments. The main disadvantages of these methods are ...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/30
CPCG01N27/30G01N27/3271
Inventor 吴韶华王芳芳孙建军张标米真真
Owner FUZHOU UNIVERSITY
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