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Electrochemical sensor and its prepn process and use

A sensor and electrochemical technology, applied in the field of DNA electrochemical sensor preparation, can solve the problems of insensitive detection of low-copy gene sequences, time-consuming and labor-intensive chromosome analysis, low sensitivity, etc., so as to accelerate the electron transfer rate and increase the amount of covalent fixation. , the effect of improving sensitivity

Inactive Publication Date: 2007-10-03
FUJIAN MEDICAL UNIV
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Problems solved by technology

[0003] At present, the clinical diagnosis methods of chronic myelogenous leukemia mainly include chromosome analysis, Southern Blot, RT-PCR and FISH and other techniques, but these methods all have certain limitations: chromosome analysis is time-consuming and laborious, and the sensitivity is low, and the sensitivity can reach up to 10 -8 mol / L; Although traditional Southern Blot detection has strong specificity, it is extremely insensitive to the detection of low-copy gene sequences. Not only is the method complex, the detection cycle is long, and it is expensive and radioactive contamination, which limits its wide clinical application; RT-PCR is cumbersome and expensive; FISH technology requires the use of a complex fluorescent color system, and its relative sensitivity is not high, and it is difficult to carry out large-scale clinical application

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  • Electrochemical sensor and its prepn process and use
  • Electrochemical sensor and its prepn process and use
  • Electrochemical sensor and its prepn process and use

Examples

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preparation example Construction

[0027] The preparation method of above-mentioned sensor

[0028] 1) Preparation of carboxylated multi-walled carbon nanotubes: After soaking the carbon nanotubes in HF acid to remove catalysts and other impurities, reflux in mixed acid (concentrated sulfuric acid: concentrated nitric acid) at 120°C to 130°C for 1 to 2 hours, Stand overnight, discard the supernatant, wash with water until neutral, freeze-dry, and set aside. After the carbon nanotubes were soaked in HF acid to remove catalysts and other impurities, they were boiled with mixed acid for a certain period of time, and their infrared spectra were measured. The results are shown in Figure 2. After boiling with the mixed acid. The carboxyl peak (v -cooH 1720cm) is gradually enhanced, indicating that the oxidation of mixed acids can indeed produce carboxyl groups.

[0029] 2) Preparation of multi-walled carbon nanotubes (MWNTS) modified glassy carbon electrodes: 1-100 mg of carboxylated multi-walled carbon nanotubes (...

Embodiment 1

[0035] Embodiment 1: An electrochemical sensor includes a glassy carbon electrode, and the surface of the glassy carbon electrode is coated with a sensitive film. The sensitive membrane consists of 0.01mg carboxylated multi-walled carbon nanotubes and b3-a2 gene (NH 2 -DNA) composition, the following NH 2 -The DNAs are all b3-a2 type genes with amino groups modified at the ends.

[0036] The preparation method of the above electrochemical sensor is to add 10 mg of carbon nanotubes into 10 milliliters of DMF, ultrasonically disperse until a uniform, light black carbon nanotube dispersion is obtained, and apply 10 microliters of carbon nanotube dispersion to the surface of the glassy carbon electrode , baked under infrared light to evaporate the solvent, and the MWNTS-COOH modified electrode was fully washed in ethanol and secondary water to remove excess carbon nanotubes. Then soak it in 30mmol / L acetic acid buffer solution containing 10mmol / L EDC to stir the reaction, wash i...

example 2

[0037] Example 2: An electrochemical sensor, including a glassy carbon electrode, the surface of the glassy carbon electrode is coated with a sensitive film. The sensitive film is composed of 0.1 mg carboxylated multi-walled carbon nanotubes and b3-a2 gene with terminal modified amino groups.

[0038] The preparation method of the above electrochemical sensor is to add 50 mg of carbon nanotubes into 10 ml of DMF, disperse ultrasonically until a uniform, light black carbon nanotube dispersion is obtained, and apply 20 microliters of the carbon nanotube dispersion to the surface of the glassy carbon electrode , baked under infrared light to evaporate the solvent, and the MWNTS-COOH modified electrode was fully washed in ethanol and secondary water to remove excess carbon nanotubes. Then soak it in 30mmol / L acetic acid buffer solution containing 10mmol / L EDC to stir the reaction, wash it with acetic acid buffer solution and place it in the solution containing 3.5μmol / L NH 2 -in ...

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Abstract

The present invention discloses one kind of electrochemical sensor and its preparation process and use. The electrochemical sensor is prepared through decorating the surface of glass-carbon electrode with open-mouthed multi-wall carbon nanotube possessing carboxylated end, fixing BCR / ABL b3-a2 type gene probe through covalent decoration onto the end carboxyl radical of carbon nanotube, hybridizing the probe DNA and the target DNA and adopting curcumin as hybridization indicator for researching the recognizing capacity of single strand fixed on the decorated electrode on complementary DNA. The carbon nanotube has great specific surface area favoring the fixing of DNA on the electrode and excellent electron transferring characteristic for high detection sensitivity of sensor. The sensor is used in detecting chronic granulocytic leukemia gene and has high selectivity and high sensitivity.

Description

Technical field: [0001] The invention relates to biochemical engineering, sensor and medical diagnosis technology, in particular to a preparation method and application of a DNA electrochemical sensor for chronic myelogenous leukemia gene detection. Background technique: [0002] Chronic myeloid leukemia, referred to as chronic myelocytic leukemia (CML), is a cell line disease caused by malignant lesions at the level of pluripotent stem cells accompanied by acquired chromosomal abnormalities. It is clinically characterized by marked granulocyte overproduction. Chronic myeloid leukemia is one of the malignant blood diseases that seriously endanger the health of young and middle-aged people, and it is a more common type of leukemia. The onset of chronic myelogenous leukemia is slow, and there are often no symptoms in the early stage, which makes clinical diagnosis difficult. The level of early diagnosis will directly affect the treatment effect and quality of life of leukemi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/403C12Q1/68
Inventor 林新华陈敬华郑艳洁陈伟张亚峰吴萍
Owner FUJIAN MEDICAL UNIV
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