Helicobacter pylori identification and virulence multiplex 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 few detection sites, 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) 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
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 identification and virulence multiplex gene detection system and its kit and application
  • Helicobacter pylori identification and virulence multiplex gene detection system and its kit and application
  • Helicobacter pylori identification and virulence multiplex gene detection system and its kit and application

Examples

Experimental program
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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

[0114] The Helicobacter pylori detection kit of this embodiment is the same as the rest of Example 1, except that the primer mixture only includes 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 luxS Forward primers for genes, targeting luxS Gene reverse primer.

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Abstract

The invention relates to a helicobacter pylori identification and virulence multiplex gene detection system, a kit adopting the detection system and the application of the detection system. The detection system comprises a plurality of pairs of primers aiming at strain identification genes (16S rRNA) and virulence genes (cagA, vacA-s1, vacA-s2, vacA-m1, vacA-m2, iceA1, iceA2, dupA, oipA and luxS) respectively. According to the helicobacter pylori identification and virulence multiplex gene detection system and the kit adopting the detection system, routine culture and other steps are not needed, helicobacter pylori identification and multiplex virulence analysis can be directly conducted on a tissue sample in the same reaction system, the defects of conventional detection methods of low flux, high time consumption and low detection rate are overcome, comprehensive, accurate and low-cost etiology basis is provided for clinical application quickly, and important reference is provided for helicobacter pylori infection accurate diagnosis and differential diagnosis and disease prognosis.

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, pregnancy vomiting, colorectal cancer and various skin diseases, and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/6851C12Q1/6858C12Q1/06C12Q1/04C12R1/01
CPCC12Q1/6851C12Q1/6858C12Q1/689C12Q2600/106C12Q2600/16C12Q2537/143C12Q2545/113C12Q2565/125
Inventor 赵虎张艳梅杨长青边海鹏胡彬婕赵付菊吴勇王诗雯缪应新徐玲丽孔咪咪张景皓姜文荣陈飞
Owner HUADONG HOSPITAL
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