Method for screening helicobacter pylori urease inhibitor

A technology for urease inhibitors and Helicobacter pylori, which is applied in the field of screening Helicobacter pylori urease inhibitors to achieve rapid detection, high detection stability, and simple preparation process

Active Publication Date: 2021-10-08
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Research and professional methods for screening Helicobacter pylori urease inhibitors based on biosensor technology are still blank

Method used

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  • Method for screening helicobacter pylori urease inhibitor
  • Method for screening helicobacter pylori urease inhibitor
  • Method for screening helicobacter pylori urease inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1: A kind of method of screening Helicobacter pylori urease inhibitor AHA

[0045] (1) Preparation and characterization of HPUb / nPt / NPG / GCE biosensor:

[0046] The NPG film prepared by corroding the Au / Ag alloy sheet with concentrated nitric acid at 40°C for 1 h was attached to the pretreated GCE; then nPt was electrodeposited on the surface of NPG / GCE by cathodic reduction plating to obtain nPt / NPG / GCE, in which The electrolyte contains 5mM H 2 PtCl 6 0.5mol / L H 2 SO 4 solution with a current density of 5mA / cm 2 , the electrodeposition time was 120s; finally, 20 μL of purified HPUb was drop-coated on the surface of the nPt / NPG / GCE, and self-assembled at 4°C for 24 hours; the HPUb / nPt / NPG / GCE biosensor was finally obtained;

[0047] The prepared HPUb / nPt / NPG / GCE was used as the working electrode, the platinum electrode was used as the counter electrode, and the saturated calomel electrode was used as the reference electrode to form a three-electrode syst...

Embodiment 2

[0060] Embodiment 2: A kind of method of screening Helicobacter pylori urease inhibitor hydroxyurea

[0061] (1) Preparation and characterization of HPUb / nPt / NPG / GCE biosensor:

[0062] The NPG film prepared by corroding the Au / Ag alloy sheet with concentrated nitric acid at 40°C for 1 h was attached to the pretreated GCE; then nPt was electrodeposited on the surface of NPG / GCE by cathodic reduction plating to obtain nPt / NPG / GCE, in which The electrolyte contains 5mM H 2 PtCl 6 0.5mol / L H 2 SO 4 solution with a current density of 5mA / cm 2 , the electrodeposition time was 120s; finally, 20 μL of purified HPUb was drop-coated on the surface of the nPt / NPG / GCE, and self-assembled at 4°C for 24 hours; the HPUb / nPt / NPG / GCE biosensor was finally obtained;

[0063] The prepared HPUb / nPt / NPG / GCE was used as the working electrode, the platinum electrode was used as the counter electrode, and the saturated calomel electrode was used as the reference electrode to form a three-electr...

Embodiment 3

[0072] Embodiment 3: A kind of method of screening Helicobacter pylori urease inhibitor methylurea

[0073] (1) Preparation and characterization of HPUb / nPt / NPG / GCE biosensor:

[0074] The NPG film prepared by corroding the Au / Ag alloy sheet with concentrated nitric acid at 40°C for 1 h was attached to the pretreated GCE; then nPt was electrodeposited on the surface of NPG / GCE by cathodic reduction plating to obtain nPt / NPG / GCE, in which The electrolyte contains 5mM H 2 PtCl 6 0.5mol / L H 2 SO 4 solution with a current density of 5mA / cm 2 , the electrodeposition time was 120s; finally, 20 μL of purified HPUb was drop-coated on the surface of the nPt / NPG / GCE, and self-assembled at 4°C for 24 hours; the HPUb / nPt / NPG / GCE biosensor was finally obtained;

[0075] The prepared HPUb / nPt / NPG / GCE was used as the working electrode, the platinum electrode was used as the counter electrode, and the saturated calomel electrode was used as the reference electrode to form a three-electro...

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Abstract

The invention discloses a method for screening a helicobacter pylori urease inhibitor by using an HPUb/nPt/NPG/GCE biosensor. The method comprises the following steps of: (1) preparing and representing a HPUb/nPt/NPG/GCE biosensor; (2) detecting a response current generated by urea with the gradient concentration of 0-100mM by using the HPUb/nPt/NPG/GCE biosensor, and calculating according to a Mie's equation to obtain K<a, urea> and V<max, urea> of the urea detected by the biosensor; and (3) detecting different urease inhibitors, and screening and analyzing inhibition types and inhibition constants of the inhibitors. The HPUb/nPt/NPG/GCE biosensor developed in the method is simple in preparation process, high in detection stability and easy to store. Meanwhile, compared with other detection methods, the screening method provided by the invention does not need a complex sample pretreatment process, is simple to operate and rapid in detection, effectively shortens the potential urease inhibitor screening time, and has a wide application prospect.

Description

technical field [0001] The present invention relates to a kind of method of screening Helicobacter pylori (Helicobacter pylori, H. pylori) urease inhibitor, relate in particular to a method utilizing H. pylori urease b subunit (H. pylori 26695 urease b subunit, HPUb) / nano platinum (Nano Platinum, nPt) / nanoporous gold (Nanoporous gold, NPG) / glassy carbon electrode (Glassy carbon electrode, GCE) biosensor screening method for Helicobacter pylori urease inhibitors. It belongs to the field of electrochemical analysis and testing. Background technique [0002] Helicobacter pylori (H. pylori) is a Gram-negative microaerophilic bacterium that selectively colonizes the surface of human gastric mucosal epithelial cells and the bottom layer of gastric mucus. It is well known that H. pylori is associated with the development of chronic gastritis, peptic ulcer, gastric mucosa-associated lymphoid tissue lymphoma and gastric adenocarcinoma. Although combination therapies based on an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327C12Q1/34C12Q1/00
CPCG01N27/3277C12Q1/34C12Q1/005G01N2333/98
Inventor 王霞肖洒
Owner SHANDONG UNIV
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