Electrochemical biosensor used for detecting lysozyme and a preparation method thereof

A technology of biosensor and lysozyme, which is applied in the field of electrochemical detection, can solve the problems of complicated operation and high cost, and achieve the effect of high sensitivity operation, low cost and simple method

Active Publication Date: 2015-10-21
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Usually, for the detection of lysozyme, lysozyme must be separated first. Common methods include gel electrophoresis, high performance liquid chromatography, immunoelectrophoresis, enzyme-linked immunosorbent assay, etc. Although these methods have high sensitivity, they have Complicated operation, high cost and many other disadvantages

Method used

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  • Electrochemical biosensor used for detecting lysozyme and a preparation method thereof
  • Electrochemical biosensor used for detecting lysozyme and a preparation method thereof
  • Electrochemical biosensor used for detecting lysozyme and a preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0046] A preparation method of an electrochemical biosensor for detecting lysozyme, such as figure 1 shown, including the following steps:

[0047] (1) Design a reporter probe DNA strand S1, which can be complementary paired with the lysozyme aptamer strand S2 and has a complementary pairing sequence of 16 bases in length, which can form a neck loop structure after the aptamer strand is detached; The 3' end of the S1 chain is modified with a ferrocene label, which can generate an electrochemical signal; the 5' end of the S1 chain is thiolated, so that S1 is modified to the surface of the gold electrode through a gold-sulfur bond; among them, the reporter probe The DNA chain sequence of S1 is: 5'-SH-C6-GTGCAGAGCTAAGTAACTCTGCAC-Fc-3', as shown in SEQ ID NO.1 in the sequence listing; the DNA chain sequence of aptamer S2 is: 5'-ATC AGG GCT AAA GAG TGC AGA GTT ACT TAG-3', as shown in SEQ ID NO.2 in the sequence listing;

[0048] (2) Use 0.05um Al for gold electrodes on chamois 2...

Embodiment 2

[0052] A preparation method of an electrochemical biosensor for detecting lysozyme, such as figure 1 shown, including the following steps:

[0053] (1) Design a reporter probe DNA strand S1, which can be complementary paired with the lysozyme aptamer strand S2 and has a complementary pairing sequence of 16 bases in length, which can form a neck loop structure after the aptamer strand is detached; The 3' end of the S1 chain is modified with a ferrocene label, which can generate an electrochemical signal; the 5' end of the S1 chain is thiolated, so that S1 is modified to the surface of the gold electrode through a gold-sulfur bond; among them, the reporter probe The DNA chain sequence of S1 is: 5'-SH-C6-GTGCAGAGCTAAGTAACTCTGCAC-Fc-3', as shown in SEQ ID NO.1 in the sequence listing; the DNA chain sequence of aptamer S2 is: 5'-ATC AGG GCT AAA GAG TGC AGA GTT ACT TAG-3', as shown in SEQ ID NO.2 in the sequence listing;

[0054] (2) Use 0.05um Al for gold electrodes on chamois 2...

Embodiment 3

[0058] The application of the electrochemical biosensor prepared in Example 1-2 in the detection of lysozyme, the steps are as follows:

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Abstract

The invention discloses a preparation method of an electrochemical biosensor used for detecting lysozyme. The preparation method includes the following steps that step1, a reporter probe DNA chain S1 is designed; step2, a gold electrode is pretreated; step3, the disulfide bond of S1 is opened in a buffering solution, and then lysozyme aptamers S2 are added so that S2 and S1 can form double-chain DNA; step4, the treated gold electrode is soaked in the double-chain DNA solution, and the double-chain DNA is arranged on the surface of the gold electrode in a modified mode. The invention further discloses the electrochemical biosensor prepared through the method and application of the sensor in the process of detecting lysozyme. The obtained electrode is soaked in hexaammineruthenium (III) chloride detection solution, then the obtained electrode, a calomel reference electrode and a platinum wire contrast electrode form a three-electrode system, and SWV detecting is performed. For the first time, a strategy of amplifying double electrochemical signals is combined with a label-free strategy to be applied to detecting the lysozyme, and therefore the lysozyme is detected in a high-sensitivity mode.

Description

technical field [0001] The invention relates to the technical field of electrochemical detection, in particular to an electrochemical biosensor for detecting lysozyme and a preparation method thereof. Background technique [0002] Lysozyme is an antibacterial protein secreted by polymorphonuclear leukocytes. Usually in the human body, lysozyme has physiological and pharmaceutical functions, such as anti-inflammatory, antiviral, immune regulation and antitumor. The concentration of lysozyme in human fluid can be used as a clinical marker for the diagnosis of many diseases, such as the concentration of lysozyme in the saliva of patients with oropharyngeal candida infection will increase; lysozyme will appear in the urine of patients with kidney disease. The concentration of lysozyme in blood and cerebrospinal fluid is an important auxiliary diagnostic basis for leukemia and central nervous system tumor diseases, so the detection of lysozyme is of great significance in clinical...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26
Inventor 夏建飞曹喜玥王宗花杨敏张菲菲
Owner QINGDAO UNIV
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