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Helicobator pylori nucleic acid sensor, detection method and application

A technology of Helicobacter pylori and nucleic acid sensor, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, and microbial determination/inspection, and can solve the problems of limiting the development of Helicobacter pylori nucleic acid detection and insufficient detection sensitivity.

Active Publication Date: 2020-10-02
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, after research by the inventors, it has been found that the nucleic acid of Helicobacter pylori has been highly diluted in both environmental samples and physiological samples, and the detection sensitivity of the prior art is insufficient, thus limiting the development of detection of Helicobacter pylori nucleic acid

Method used

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  • Helicobator pylori nucleic acid sensor, detection method and application
  • Helicobator pylori nucleic acid sensor, detection method and application
  • Helicobator pylori nucleic acid sensor, detection method and application

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Embodiment approach

[0056] The third embodiment of the present invention provides an application of the above-mentioned Helicobacter pylori nucleic acid sensor in the preparation of a reagent for detecting Helicobacter pylori.

[0057] The fourth embodiment of the present invention provides a Helicobacter pylori detection kit, comprising the above-mentioned Helicobacter pylori nucleic acid sensor, a buffer solution, and deoxyribonucleoside triphosphate (dNTP).

[0058] In order to enable those skilled in the art to understand the technical solution of the present invention more clearly, the technical solution of the present invention will be described in detail below in conjunction with specific embodiments.

Embodiment 1

[0060] Synthesis of CdS quantum dot solution.

[0061] At 70°C with constant stirring, 0.1937g of sodium sulfide (Na 2 S) Na dissolved in 30mL ultrapure water (newly prepared) 2 S solution was slowly added to 0.1861g cadmium nitrate tetrahydrate (Cd(NO) 3 4H 2 O) dissolved in 30 mL of ultrapure water to obtain an orange-yellow solution, refluxed at 70° C. for 3 hours, and cooled to room temperature. Transfer the solution to a centrifuge tube and centrifuge at 15,000r / min for 10min, collect the precipitate, wash once with ethanol and ultrapure water, remove the supernatant by centrifuging at 15,000 rpm after each wash, collect the precipitate, add pure water for ultrasonic dispersion, and then 12,000 rpm After centrifugation, the supernatant collected is the CdS quantum dot solution, and the fluorescence spectrum of the obtained CdS quantum dots is as follows: figure 2 As shown in A, TEM as figure 2 Shown in B.

[0062] Modifications and reaction processes on electrodes...

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Abstract

The invention discloses a helicobator pylori nucleic acid sensor, a detection method and application. The sensor comprises a glassy carbon electrode, a primer sequence, a template sequence, a DNA ligase, a DNA polymerase, a water-soluble silver salt and a reducing agent; the primer sequence and CdS quantum dots are connected with the surface of the glassy carbon electrode; the template sequence can form a dumbbell-shaped structure, and the dumbbell-shaped structure can be connected with the DNA ligase to be closed by the DNA ligase; and the template sequence, the primer sequence and a helicobator pylori nucleic acid sequence can form a trimer structure, the trimer structure can make the primer sequence subjected to rolling circle amplification under the action of the DNA polymerase, and anamplified DNA long chain has repetitive sequences rich in cytosine bases. By means of the helicobator pylori nucleic acid sensor, the detection method and application, ultra-sensitive detection of helicobator pylori nucleic acid can be achieved.

Description

technical field [0001] The invention relates to a helicobacter pylori nucleic acid sensor, a detection method and an application. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Helicobacter pylori is a Gram-negative bacterium and one of the most common bacterial pathogens. It mainly lives in the pylorus of the human stomach and is a potential cause of many diseases such as gastritis, peptic ulcer, and lymphoproliferative gastric lymphoma. 67%-80% of gastric ulcers and 95% of duodenal ulcers are thought to be caused by H. pylori. In recent years, Helicobacter pylori infection has been found to be closely related to gastric cancer, and the World Health Organization define...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/6844C12Q1/04C12R1/01
CPCC12Q1/689C12Q1/6844C12Q2521/101C12Q2521/501C12Q2531/125C12Q2563/103C12Q2565/607
Inventor 胡善文张晓荣郭睿吴衍王文祥
Owner FUJIAN MEDICAL UNIV
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