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Helicobacter pylori qualitative and quantitative multiple gene detection system and its kit and application

A Helicobacter pylori and gene detection technology, applied in the biological field, can solve the problems of not reflecting the current infection, unable to do quantitative analysis, few detection sites, etc., to guide accurate diagnosis and differential diagnosis and curative effect observation, low cost, highly specific effect

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

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Problems solved by technology

Its disadvantage is that it does not reflect current infection
5) Nucleic acid analysis methods: including sequencing, PCR, oligonucleotide probe hybridization, etc., but these inspection methods have few detection sites, low specificity, low throughput, high cost, and cannot be used for quantitative analysis
6) Quantitative analysis: The commonly used quantitative analysis method is real-time PCR, but this method has a single detection, low throughput, and high cost when analyzing multiple genes

Method used

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

Examples

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Embodiment 1

[0050] 1. The composition of the kit

[0051] 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.

[0052] PCR buffer, dNTPs and hot-start DNA polymerase were all from Takara (Catalog No.: R007A).

[0053] The positive control solution is a plasmid mix that includes all gene targets of interest.

[0054] The negative control solution was nuclease-free ultrapure water.

[0055] 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 primers for genes, t...

Embodiment 2

[0115] The Helicobacter pylori detection kit of this embodiment is the same as the rest of Example 1, except that the primer mixture only includes 16S rRNA Forward primers for genes, targeting 16S rRNA Reverse primers for genes, targeting ureC Forward primers for genes, targeting ureC Reverse primers for genes, targeting ß-globin Forward primers for genes, targeting ß-globin Gene reverse primer.

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Abstract

The invention relates to a qualitative and quantitative multiple-gene helicobacter pylori detection system and a kit and application thereof. The qualitative and quantitative multiple-gene helicobacter pylori detection system includes multiple pairs of primers respectively for a strain identification gene (16S rRNA) and quantitative analysis genes ureC and beta-globin of the helicobacter pylori. The qualitative and quantitative multiple-gene helicobacter pylori detection system and the kit of the system do not need the steps including conventional isolated culture of bacteria and the like, strain identification and quantitative analysis can be directly conducted on tissue samples in the same reaction system, the accuracy of detection results is obviously improved, an accurate and low-cost etiological basis is clinically provided for the first time, and important references are provided for accurate diagnosis and differential diagnosis of helicobacter pylori infection and curative effect observation.

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): C12Q1/689C12Q1/6851C12Q1/6858C12Q1/06C12Q1/04
CPCC12Q1/6851C12Q1/6858C12Q1/689C12Q2600/106C12Q2600/16C12Q2537/143C12Q2545/113C12Q2565/125
Inventor 张艳梅赵虎项平胡彬婕赵付菊周丽芳吴勇王诗雯缪应新方毅季大年黄任翔陈洁徐玲丽孔咪咪张景皓姜文荣陈飞
Owner HUADONG HOSPITAL
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