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Multichannel Fibrin detection method of nanostructure artificial enzyme signal probe

A technology of signal probes and nanostructures, applied in the field of medical testing, can solve the problems of uncontrollable interfaces, low enzyme reaction efficiency, and difficult disorder, etc., achieve strong penetration and sensitivity, and improve detection sensitivity , the effect of good stability

Active Publication Date: 2020-06-30
SHANGHAI UNIV OF MEDICINE & HEALTH SCI
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Problems solved by technology

Horseradish peroxidase is one of the most commonly used amplification signal enzymes for electrochemical biosensors due to its good stability, high efficiency, and commercialization. However, due to its uncontrollable and disordered properties on the interface It is not easy to control and the enzyme reaction efficiency is not high, so there is an urgent need for an artificial enzyme with good stability and strong controllability

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  • Multichannel Fibrin detection method of nanostructure artificial enzyme signal probe
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  • Multichannel Fibrin detection method of nanostructure artificial enzyme signal probe

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Embodiment Construction

[0029] The invention provides a multi-channel Fibrin detection method of a nanostructured artificial enzyme signal probe, specifically:

[0030] 1.1 Instruments and reagents

[0031] Gradient PCR instrument (Thermal Cycler T100TM, BIO-RAD), 16-channel electrochemical biosensing system (Zhejiang Nazhihui Biotechnology Co., Ltd.)

[0032] Homing peptide (CREKA, Shanghai Qiangyao Biotechnology Co., Ltd.), 3,3',5,5'-tetramethylbenzidine (TMB, Neogen), 1-ethyl-(3-dimethylaminopropyl ) carbonate diimide hydrochloride (EDC, Sinopharm Chemical Reagent Co., Ltd.), N-hydroxysuccinimide (NHS, Shanghai Aladdin Biochemical Technology Co., Ltd.), hemin (hemin, Shanghai Biological Biotechnology Co., Ltd. Co., Ltd.), fibrinogen (Fibrinogen, Shanghai Biological Biotechnology Co., Ltd.), thrombin (Thrombin, Shanghai Titan Technology Co., Ltd.), Gel Red (Shanghai Liji Biotechnology Co., Ltd.), 10 × PBS buffer (10mM Na 2 HPO 4 , 2mM KH 2 PO 4 , 37mM NaCl and 2.7mM KCl, pH7.4), 10×TMK buffer...

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Abstract

The present invention discloses a multichannel Fibrin detection method of a nanostructure artificial enzyme signal probe. A novel artificial enzyme synthesized by a DNA tetrahedron three-dimensional probe is used for detecting fibrin, and has high stability and relatively strong penetrativity and sensitivity; DNA as a multifunctional biological nanomaterial has the outstanding application in the fields of nanostructure self-assembling, biological immune sensors and the like; and the DNA tetrahedron three-dimensional probe as a skeleton of the artificial enzyme provides a stable chemical environment for hemin / G4-DNAzyme, thereby constructing the novel artificial enzyme (Tetrazyme). The artificial enzyme can efficiently detect the fibrin by only the DNA with the concentration being 0.6 [mu]Mand CREKA with the concentration being 80 [mu]M. A result shows that the novel electrochemical detection method can efficiently and conveniently detect the fibrin and is high in sensitivity.

Description

technical field [0001] The invention relates to the technical field of medical detection, in particular to a multi-channel Fibrin detection method of a nanostructured artificial enzyme signal probe. Background technique [0002] Tumor is one of the main diseases that threaten people's life and health, and the incidence of malignant tumors is increasing year by year, so the efficient screening, diagnosis and treatment of tumor cells has become a key factor, and the detection of fibrin plays a role in the early screening of cancer cells Significantly, studies have shown that the interstitium of malignant tumor cells contains fibrin with a high concentration of fibrous network structure, which can increase the permeability of blood vessels and form fibrin in the interstitium with plasma protein leakage, thus giving cancer Good conditions are provided for the growth and transfer of cells. Currently, the methods for detecting fibrin are mainly fluorescent imaging, latex agglutin...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/44G01N27/30G01N27/327G01N27/48
CPCC12Q1/44G01N27/308G01N27/3277G01N27/3278G01N27/48G01N2333/916Y02A50/30
Inventor 曾冬冬王彬徐晓慧李晖潘洪志
Owner SHANGHAI UNIV OF MEDICINE & HEALTH SCI
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