Helicobacter pylori multiple gene detection system and its kit and application

A technology for Helicobacter pylori and gene detection, which is applied in the directions of biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of relying on isolated and cultured strains, cumbersome operations, and low throughput, so as to guide individualized medicine and curative effect. Monitor, avoid mutual interference, compensate for low throughput effects

Active Publication Date: 2019-02-26
HUADONG HOSPITAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its disadvantage is that it does not reflect current infection
5) Quantitative analysis: real-time PCR is commonly used, but this method has a single detection, low throughput, and high cost when analyzing multiple genes
6) Drug susceptibility test: This method is time-consuming, cumbersome to operate, and relies on the isolation and culture of strains
However, the current conventional detection methods have shortcomings such as long time, low sensitivity, high cost, low throughput, and in particular, the inability to detect multiple related factors at the same time.

Method used

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  • Helicobacter pylori multiple gene detection system and its kit and application
  • Helicobacter pylori multiple gene detection system and its kit and application
  • Helicobacter pylori multiple gene detection system and its kit and application

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Embodiment

[0051] 1. The composition of the kit

[0052] The Helicobacter pylori detection kit of this example includes: primer mixture, PCR buffer (10×PCRBuffer), MgCl 2 solution, dNTPs, fluorescent Universal Labeling Mix, hot-start DNA polymerase (Taq DNA polymerase), positive and negative controls. 2mmol / L of dNTPs and fluorescent universal tag mix are mixed together into one tube of reagents.

[0053] PCR buffer, dNTPs and hot-start DNA polymerase were all from Takara (Catalog No.: R007A).

[0054] The positive control solution is a plasmid mix that includes all gene targets of interest.

[0055] The negative control solution was nuclease-free ultrapure water.

[0056] The primer mix includes 16S rRNA Forward primers for genes, targeting 16S rRNA Reverse primers for genes, targeting cagA Forward primers for genes, targeting cagA Reverse primers for genes, targeting vacA-s1 or vacA-s2 Forward primers for genes, targeting vacA-s1 or vacA-s2 reverse primer for gene va...

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Abstract

The invention relates to a multiple-gene helicobacter pylori detection system and a kit and application thereof. The helicobacter pylori detection system includes 21 pairs of primers respectively for a strain identification gene (16S rRNA), virulence genes (cagA, vacA-s1, vacA-s2, vacA-m1, vacA-m2, iceA1, iceA2, dupA, oipA and luxS), drug resistance genes (2143 locus of 23S rRNA, 148 locus of rdxA, 1777 locus of pbp1A and polymorphism of the 261 locus of gyrA) and quantitative analysis genes ureC and beta-globin of the helicobacter pylori. The multiple-gene helicobacter pylori detection system and the kit of the system do not need the steps including conventional isolated culture and the like, synchronous detection and analysis on strain identification, quantification, virulence and medicine resistance can be directly conducted on tissue samples in the same reaction system, the shortcomings of low flux, long consumed time, low detection rate and the like of a conventional detection method are overcome, a comprehensive, accurate and low-cost etiological diagnosis is clinically provided for the first time, and important references are provided for individualized diagnosis and accurate treatment of helicobacter pylori infection.

Description

technical field [0001] The invention relates to a multiple gene detection product and a detection system used in the product, belonging to the field of biotechnology. Background technique [0002] Helicobacter pylori ( H. pylori ) is a gram-negative, microaerophilic, campylobacter that primarily inhabits the human stomach. Helicobacter pylori infection is closely related to the occurrence and development of chronic atrophic gastritis, peptic ulcer, gastric mucosa-associated lymphoid tissue lymphoma and gastric cancer, so it has attracted widespread clinical attention. In 1994, the International Agency for Research on Cancer (IARC) listed it as a human I carcinogen, and it is the only bacterial pathogenic microorganism listed as a clear carcinogen to humans so far. Many reports believe that Helicobacter pylori infection is related to coronary heart disease, rheumatoid, hepatobiliary disease, tuberculosis, vomiting of pregnancy, colorectal cancer and various skin diseases. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/6851C12Q1/6858C12Q1/04C12Q1/06
CPCC12Q1/6851C12Q1/6858C12Q1/689C12Q2600/106C12Q2600/16C12Q2537/143C12Q2545/113C12Q2565/125
Inventor 赵虎张艳梅保志军杨长青项平胡彬婕赵付菊陈洁周丽芳吴勇王诗雯缪应新方毅季大年黄任翔徐玲丽边海鹏南丽余丁张景皓姜文荣陈飞
Owner HUADONG HOSPITAL
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