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

A technology for Helicobacter pylori and gene detection, applied in the biological field, can solve the problems of low sensitivity, low throughput, and long time, and achieve the effects of avoiding mutual interference, compensating for low throughput, and reducing false positives

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
However, the current conventional detection methods have shortcomings such as long time, low sensitivity, high cost, low throughput, and inability to detect multiple related factors at the same time.

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

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-s2Forward primers for genes, targeting vacA-s1 or vacA-s2 Reverse primers for genes, ...

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 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, targeting vacA-m1 Forward primers for genes, targeting vacA-m1 Reverse primers for genes, targeting vacA-m2 Forward primers for genes, targeting vacA-m2 Reverse primers for genes, targeting iceA1 Forward primers for genes, targeting iceA1 Reverse primers for genes, targeting iceA2 Forward primers for genes, targeting iceA2 Reverse primers for genes, targeting dupA Forward primers for genes, targeting dupA Reverse primers for genes, targeting oipA Forward primers for genes, targeting oipA Reverse primers for genes, targeting ...

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Abstract

The invention relates to a system for detecting multiple quantitative and virulent genes of H.pylori as well as kits and applications of the system. The system for detecting multiple quantitative and virulent genes of H.pylori comprises multiple pairs of primers, each for strain identification genes (16S rRNA), quantitative analysis genes ureCand Beta-globin, as well as virulent genes (cagA, vacA-s1, vacA-s2, vacA-m1, vacA-m2, iceA1, iceA2, dupA, oipA and luxS). The system for detecting multiple quantitative and virulent genes of H.pylori and the kits of the system can directly perform synchronous detection and analysis on the strain identification, quantification and virulence of a tissue sample in a same reaction system without adopting a conventional culture step or other steps, remedy defects that a conventional detection method is low in throughput, time-consuming and low in detection rate, obviously improve the accuracy of detection results, immediately provide a comprehensive, accurate and low-cost etiological diagnosis for clinic, and provide an important reference for the accurate diagnosis, differential diagnosis and disease prognosis of H.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 diseases such as coronary heart disease, rheumatoid, hepatobiliary disease, tuberculosis, pregnancy vomiting, colorectal cancer and various sk...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12N15/11
CPCC12Q1/6851C12Q1/689C12Q2600/16
Inventor 张艳梅赵虎赵付菊胡彬婕王诗雯陈飞吴勇缪应新张景皓姜文荣徐玲丽南丽
Owner HUADONG HOSPITAL
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