Self-assembled label-free magnetic nano CYP2C9 * 3 gene probe and preparation method thereof

A magnetic nanometer and gene probe technology, applied in the field of biological analysis, can solve the problems of high detection limit, expensive requirements, application limitations, etc., and achieve the effect of fast detection process, less probe amount and good sensitivity

Pending Publication Date: 2020-11-13
GUODIAN SCI & TECH RES INST +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] CYP2C9*3 genetic testing projects have been carried out clinically, but due to the relatively high price, the application is greatly limited
Traditional genetic testing methods are time-consuming, costly, have high detection limits, and require expensive instruments

Method used

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  • Self-assembled label-free magnetic nano CYP2C9 * 3 gene probe and preparation method thereof
  • Self-assembled label-free magnetic nano CYP2C9 * 3 gene probe and preparation method thereof
  • Self-assembled label-free magnetic nano CYP2C9 * 3 gene probe and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Fe of the present invention 3 o 4 / Fe 2 o 3 Preparation of @Au-PNA / MCH / DNA nanometer CYP2C9*3 gene probe:

[0052] 1) will prepare the Fe prepared by the example 3 o 4 / Fe 2 o 3 The @Au nanocomposite was ultrasonically dispersed in ultrapure water and formulated to a concentration of 15 mg / mL Fe 3 o 4 / Fe 2 o 3 @Au nanocomposite suspension, spare;

[0053] 2) Dispense 10 µL of 15 mg / mL Fe 3 o 4 / Fe 2 o 3 The @Au nanocomposite suspension was added dropwise to the surface of the magnetic glassy carbon electrode (MGCE). After the liquid dried, a uniform layer of Fe was formed on the surface of the electrode. 3 o 4 / Fe 2 o 3 @Au Membrane.

[0054] 3) Mix TCEP at a concentration of 48 µM with an aqueous solution of thiol-modified PNA at a concentration of 1.6 µM to prepare a thiol-modified PNA solution containing TCEP for use;

[0055] 4) Add 5 µL of the thiol-modified PNA solution containing TCEP in 3) to the electrode surface dropwise, 4 o C for 12 h. ...

Embodiment 2

[0066] Fe of the present invention 3 o 4 / Fe 2 o 3 Preparation of @Au-PNA / MCH / DNA nanometer CYP2C9*3 gene probe:

[0067] 1) The Fe prepared by the above preparation example 3 o 4 / Fe 2 o 3 The @Au nanocomposite was ultrasonically dispersed in ultrapure water and formulated to a concentration of 13 mg / mL Fe 3 o 4 / Fe 2 o 3 @Au nanocomposite suspension;

[0068] 2) Add 10 µL of 13 mg / mL Fe 3 o 4 / Fe 2 o 3 The @Au nanocomposite suspension was added dropwise to the surface of the magnetic glassy carbon electrode (MGCE). After the liquid dried, a uniform layer of Fe was formed on the surface of the electrode. 3 o 4 / Fe 2 o 3 @Au Membrane.

[0069] 3) Mix 48 µM TCEP with 0.8 µM thiol-modified PNA aqueous solution to prepare a thiol-modified PNA solution containing TCEP.

[0070] 4) Take 5 µL of the thiol-modified PNA solution containing TCEP and drop it on the electrode surface, 4 o C for 12 h.

[0071] 5) Use 3 µL with a concentration of 1 mM MCH to block non...

Embodiment 3

[0075] Fe of the present invention 3 o 4 / Fe 2 o 3 Preparation of @Au-PNA / MCH / DNA nanometer CYP2C9*3 gene probe:

[0076] 1) The Fe prepared by the above preparation example 3 o 4 / Fe 2 o 3 @Au nanocomposites were ultrasonically dispersed in ultrapure water and formulated to a concentration of 5 mg / mL Fe 3 o 4 / Fe 2 o 3 @Au nanocomposite suspension;

[0077] 2) Add 10 µL of 5 mg / mL Fe 3 o 4 / Fe 2 o 3 The @Au nanocomposite suspension was added dropwise to the surface of the magnetic glassy carbon electrode (MGCE). After the liquid dried, a uniform layer of Fe was formed on the surface of the electrode. 3 o 4 / Fe 2 o 3 @Au Membrane.

[0078] 3) Mix 48 µM TCEP with 1.6 µM thiol-modified PNA aqueous solution to prepare a thiol-modified PNA solution containing TCEP for later use.

[0079] 4) Take 5 µL of the thiol-modified PNA solution containing TCEP and drop it on the electrode surface, 4 o C for 12 h.

[0080] 5) Use 3 µL with a concentration of 1 mM MCH to...

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Abstract

The invention provides a self-assembled label-free magnetic nano CYP2C9 * 3 gene probe and a preparation method thereof. The probe is a Fe3O4 / Fe2O3(at)Au-PNA / MCH / DNA nano CYP2C9 * 3 gene probe. The linear range of the detectable concentration of the probe is 1 pM-1 mu M, the peak current range is 55-89 mu A, the detection limit is 0.95 pM (S / N = 3), and the quantification limit is 3.18 pM (S / N = 10). The nano DNA probe prepared by the invention is simple and rapid to operate, strong in specificity, short in detection period, low in required equipment price and strong in applicability.

Description

technical field [0001] The invention relates to a self-assembly label-free magnetic nanometer CYP2C9*3 gene probe and a preparation method thereof, belonging to the technical field of biological analysis. Background technique [0002] Since the American medical community put forward the concept of "precision medicine" in 2011, genetic testing has received high attention. The so-called "individualized medicine" based solely on age, gender, and health status is far from meeting medical needs. Genetic factors are the source of individual differences in drug use, and the "tailor-made" drug use based on medical diagnosis and genomics has become a new medical model with a higher scientific nature. The key to precise clinical diagnosis, prevention and treatment is the selection of precise molecular markers. [0003] Cytochrome P450 (CYP450) is an important enzyme involved in drug metabolism, genetic differences in its expression and function lead to differences in the absorption ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/10C12Q1/6858C12Q1/682C12Q1/6883G01N27/327
CPCC12Q1/6858C12Q1/682C12Q1/6883G01N27/3277C12Q2600/106
Inventor 许月阳杨希刚刘瑞江薛建明王宏亮管一明陈书建董月红李忠华丁铭张强
Owner GUODIAN SCI & TECH RES INST
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