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Manufacturing method and application of electrochemiluminescence sensor for detecting thrombin

An electrochemical and sensor technology, which is applied in the field of preparation of electrochemiluminescent sensors, can solve the problem of low sensitivity of the method, and achieve the effect of high sensitivity and good development prospects

Active Publication Date: 2012-06-20
武汉菲思特医学检验实验室有限公司
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  • Application Information

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Problems solved by technology

These methods have their own advantages and can meet the detection requirements of thrombin to varying degrees, but the sensitivity of the method is not high

Method used

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  • Manufacturing method and application of electrochemiluminescence sensor for detecting thrombin
  • Manufacturing method and application of electrochemiluminescence sensor for detecting thrombin
  • Manufacturing method and application of electrochemiluminescence sensor for detecting thrombin

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preparation example Construction

[0041] The present invention is achieved through the following measures: a preparation method of an electrochemiluminescence sensor for detecting thrombin, comprising the following steps:

[0042] (1) Utilize existing method, prepare the gold nanoparticle of suitable particle size;

[0043] (2) Utilize the effect of thiol and gold to modify the surface of gold nanoparticles with alkane with thiol at one end and carboxyl at the other end;

[0044] (3) Using the interaction of carboxyl and amino groups, the aminated bipyridyl ruthenium is connected to the alkane-modified gold nanoparticles to prepare an electrochemiluminescence probe;

[0045] (4) Using the specific recognition of the target and the aptamer, the cleavage of Nb.BbvCI nickase and the polymerization of DNA polymerase to prepare a DNA hybridization solution;

[0046] (5) Using hybridization technology to construct an electrochemiluminescence sensor.

[0047] The preparation of the gold ...

Embodiment

[0060] Example: Detection of thrombin by labeling an electrochemiluminescent sensor based on enzyme-cleavage cycle signal amplification technology

[0061] 1. Experimental part

[0062] 1.1 Instruments and reagents

[0063] MPI-E Electrochemiluminescence Analysis System (Xi'an Ruimai Analytical Instrument Co., Ltd.); THZ-82A Air Bath Constant Temperature Oscillator (Quantan Medical Instrument Factory); CHI660B Electrochemical Workstation (Shanghai Chenhua Instrument Company); Anke -TGL-16C Feiweng brand high-speed centrifuge (Shanghai Anting Scientific Instrument Factory); PHS-3D acidity meter (Shanghai Leici Instrument Factory); the experiment uses a three-electrode system: gold electrode and modified gold electrode are working electrodes, The Ag / AgC1 (saturated KCl) electrode is the reference electrode, and the platinum wire electrode is the counter electrode.

[0064] Mercaptohexanol (HS-(CH 2 ) 6 -OH) was purchased from Aldrich; NaH 2 PO 4 2H 2 O was...

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Abstract

The invention relates to a manufacturing method and application of an electrochemiluminescence sensor for detecting thrombin. Ruthenium bipyridine with active group is modified to the gold nanoparticle surface to form an electrochemiluminescence nanoparticle probe, which plays the role of amplifying electrochemiluminescence, and the upper part DNA single strand is modified on the working electrode surface to constitute an electrochemiluminescence sensor. The target analyte of thrombin is subjected to reaction with partial DNA double strands and then with DNA polymerase, Nb.BbvCI nicking enzyme and partial DNA single strand to obtain DNA hybrid solution which is added into the sensor, so as to increase electrochemiluminescence nanoparticle probes on the electrode surface and further lead to an enhanced electrochemiluminescence signal, and accordingly thrombin determination is realized. The inventive sensor has high selectivity and detection sensitivity.

Description

technical field [0001] The invention belongs to the field of analytical chemistry and electrochemiluminescence sensors, in particular to a preparation method and application of an electrochemiluminescence sensor for detecting thrombin. In addition, the present invention also relates to a method for detecting thrombin by using the electrochemiluminescent sensor. [0002] Background technique [0003] Thrombin (Thombin), as an important protease, plays an important role in physiological and pathological processes such as blood coagulation, inflammation and wound healing. Because patients with malignant tumors often have different degrees of bleeding tendencies, tumor tissue invasion to surrounding tissues, tumor cell metastasis, and changes in tumor tissue and tumor patient blood itself can all change the coagulation mechanism of patients. One of the important indicators to measure the coagulation mechanism is to detect the concentration of thrombin, which is of grea...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N21/76
Inventor 混旭陈怀成王卫
Owner 武汉菲思特医学检验实验室有限公司
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