Preparation method of electrochemical sensor for VKORC1-1639G>A gene polymorphism detection

A gene polymorphism, electrochemical technology, applied in the field of electrochemical detection, can solve the problem that it cannot be used as redox electricity, and achieve the effect of promoting development, highly sensitive detection, and improving biocompatibility

Inactive Publication Date: 2017-07-25
SHIYAN TAIHE HOSPITAL
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  • Abstract
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Problems solved by technology

However, nucleic acid molecules cannot be used as redox couples in biological assay reactions due to their inert nature.

Method used

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  • Preparation method of electrochemical sensor for VKORC1-1639G>A gene polymorphism detection
  • Preparation method of electrochemical sensor for VKORC1-1639G>A gene polymorphism detection
  • Preparation method of electrochemical sensor for VKORC1-1639G>A gene polymorphism detection

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Embodiment １

[0044] combined with figure 1 As shown, further described as follows:

[0045] Step 1. Dissolve 1.0 mg of amino-modified polyamide dendrimer (PAMAM) in 1 mL of double-distilled water, then add 4 mL of ethanol and 1 mL of fullerene toluene solution with a concentration of 1.5 mM to the above solution, and stir at room temperature for 36 h. Obtain a fullerene suspension with a large amount of amino groups modified on the surface, then centrifuge at 5000 rpm for 10 min, wash with absolute ethanol and double distilled water for 3 times, and redisperse the resulting product in 2 mL of 0.1 M phosphate buffer solution to obtain PAMAM -C 60 NPs, standby; the composition of the phosphate buffer solution: a concentration of 0.1M Na 2 HPO 4 , a concentration of 0.1 M KH 2 PO 4 , KCl with a concentration of 0.1 M, and the rest are deionized water; pH = 7.4;

[0046] Step 2. Put 5.6 mg cobalt chloride hexahydrate (CoCl 2· 6H 2 O) and 100 mg of polyvinylpyrrolidone (PVP) were dissol...

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Abstract

The invention provides a preparation method of an electrochemical sensor for VKORC1-1639G> A gene polymorphism detection, first, amino-modified polyamide dendritic polymer PAMAM is used to process and functionalize hollow nano platinum to obtain a tPNPs-PAMAM composite material, the tPNPs-PAMAM composite material, a single strand DNA signal probe and fullerene nanoparticles are mixed to obtain a redox probe, and by layer by layer self-assembly of reductive graphene oxide tetraethylenepentamine, nano gold and avidin, biotinylated tetrahedral DNA capture probe is fixed, and then the electrochemical sensor for VKORC1-1639G> A gene polymorphism detection is prepared. The electrochemical sensor is successfully used for detection of single base mutation of VKORC1. The electrochemical sensor has the advantages of high sensitivity, strong specificity, rapid detection and convenience. The electrochemical sensor provides a new detection method for warfarin individualized medication.

Description

technical field [0001] The invention relates to a method for preparing an electrochemical sensor for the detection of VKORC1-1639G>A gene polymorphism, especially a sandwich-type biosensor prepared based on a fullerene nanocomposite material as a redox probe for detecting VKORC1- 1639G>A gene polymorphism belongs to the field of electrochemical detection. Background technique [0002] Warfarin is the most widely used anticoagulant and is used to treat stroke, deep vein thrombosis, pulmonary embolism, or severe coronary thromboembolic disease. However, the predicted dose of warfarin for each patient may be over or under due to the lack of individual genetic information of the patient. Considering the narrow therapeutic index of warfarin, there are large individual differences in the dosage. Therefore, it is necessary to precisely adjust the dosage of warfarin according to individual differences. This difference in drug use is determined by the target site of warfarin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
CPCG01N27/3278
Inventor 袁国林王磊张吉才毛达勇王芳
Owner SHIYAN TAIHE HOSPITAL
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