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Electrochemiluminescence detection method for tumor necrosis factor alpha and kit of electrochemiluminescence detection method

A technology of tumor necrosis factor and electrochemistry, which is applied in the direction of chemiluminescence/bioluminescence, electrochemical variables of materials, and analysis by making materials undergo chemical reactions, etc. It can solve the problems of less research, low ECL intensity, and unclear luminescence mechanism, etc. question

Inactive Publication Date: 2019-01-08
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, although the fluorescence properties of AuNCs have been widely studied and applied, due to the limitations of its low ECL intensity, low quantum yield, and unclear luminescence mechanism, the research on AuNCs in the field of ECL is still relatively small.

Method used

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  • Electrochemiluminescence detection method for tumor necrosis factor alpha and kit of electrochemiluminescence detection method
  • Electrochemiluminescence detection method for tumor necrosis factor alpha and kit of electrochemiluminescence detection method
  • Electrochemiluminescence detection method for tumor necrosis factor alpha and kit of electrochemiluminescence detection method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Add 0.6 mL of 0.5 mol / L NaOH and 0.4 mL of 20 mg / mL chloroauric acid solution into 4 mL of 0.08 mol / L N-acetyl-L-cysteine ​​solution, mix well and place at 37 °C for constant temperature Incubate for 3 hours in the sink. After the reaction was completed, the reaction solution was dialyzed and purified for 24 h with a dialysis bag with a molecular weight of 3000 to obtain the purified N-acetyl-L-cysteine-protected gold nanocluster solution, which was stored in a refrigerator at 4 °C in the dark.

Embodiment 2

[0050] Glassy carbon electrodes with a diameter of 3 mm were coated with 1.0 μm, 0.3 μm, 0.05 μm Al 2 o 3 The powder is polished and polished in turn until it reaches a smooth mirror surface, and then put in HNO in turn 3 solution, absolute ethanol, ultrasonic cleaning in deionized water for 3 minutes, N 2 blow dry. Take 5 μL of the N-acetyl-L-cysteine-protected gold nanocluster solution prepared in Example 1 and drop it on the surface of the treated glassy carbon electrode, dry it at room temperature, and further soak the electrode in 0.1 mol / L React in sodium borohydride solution for 5 minutes to obtain gold nanocluster probe modified electrode. Insert the above electrodes into 0.1 mol / L pH 7.4 phosphate buffer solution containing 0.1 mol / L potassium persulfate and 0.1 mol / L KCl. The step pulse method was adopted, the initial potential was 0 V, the pulse time was 10 s, the end potential was -2 V, and the pulse time was 1 s. The high voltage of the photomultiplier tube w...

Embodiment 3

[0052] Glassy carbon electrodes with a diameter of 3 mm were coated with 1.0 μm, 0.3 μm, 0.05 μm Al 2 o 3 The powder is polished and polished in turn until it reaches a smooth mirror surface, and then put in HNO in turn 3 solution, absolute ethanol, ultrasonic cleaning in deionized water for 3 minutes, N 2 blow dry. Take 5 μL of the N-acetyl-L-cysteine-protected gold nanocluster solution prepared in Example 1 and drop it on the surface of the treated glassy carbon electrode, dry it at room temperature, and further soak the electrode in 0.1 mol / L react in sodium borohydride solution for 5 minutes to obtain a gold nanocluster probe modified glassy carbon electrode. A three-electrode system was adopted, with gold nanocluster probe-modified glassy carbon electrode as the working electrode, platinum wire electrode as the counter electrode, and Ag / AgCl as the reference electrode. 4 -H 2 SO 4 (1:1 V / V) solution, manganese dioxide was electrodeposited, the deposition potential w...

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Abstract

The invention discloses an electrochemiluminescence detection method for a tumor necrosis factor alpha and a kit of the electrochemiluminescence detection method. According to the method, the tumor necrosis factor alpha is adopted as a detection target, a high quantum yield gold nano cluster electrochemiluminescence technique and an immunoassay technique are organically combined, a high quantum yield gold nano cluster electrochemiluminescence probe is prepared by using a reduction method, a manganese dioxide nano material is adopted as an electrochemiluminescence quenching agent, an electrochemiluminescence signal is recovered through a redox reaction between ascorbic acid generated from enzyme-linked immunosorbent assay and manganese dioxide, and the method is a high-performance electrochemiluminescence tumor necrosis factor alpha detection method based on the high quantum yield gold nano cluster probe. The method has a linear range of 0.06-31pg / mL for tumor necrosis factor alpha detection, and has a detection limit of 36fg / mL. The method has the characteristics of rapidness, accuracy, high sensitivity, good selectivity, good stability, small sample use amount and the like, and has relatively good clinical application prospects.

Description

technical field [0001] The invention relates to an electrochemiluminescent tumor necrosis factor alpha detection method and a detection kit based on a gold nano-cluster probe with a high quantum yield, belonging to the field of analytical chemistry and nanotechnology. Background technique [0002] Tumor necrosis factor α (Tumor necrosis factor, TNF-α) is a non-glycosylated cytokine produced by activated macrophages that can cause hemorrhagic necrosis of various tumors. Tumor necrosis factor α not only has anti-tumor activity, but also has immune regulation, anti-virus, and promotion of cell proliferation and differentiation. However, tumor necrosis factor α is also an endogenous pyrogenic source, and it plays a major role in cerebral malaria and septic shock, endangering the lives of patients. Studies have shown that the concentration of tumor necrosis factor α in serum is related to the pathological process of many diseases, for example, neonatal listeriosis, meningococcem...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N27/30
CPCG01N21/76G01N27/30
Inventor 彭花萍黄种南吴伟华陈伟刘银环
Owner FUJIAN MEDICAL UNIV
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