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System for detecting multiple drug-resistant and quantitative genes of H.pylori as well as kits and applications of system

A technology for Helicobacter pylori and gene detection, applied in the biological field, can solve the problems of relying on isolated and cultured strains, failing to reflect current infection, and single detection, and achieve low-cost etiological diagnosis, good convenience, and high specificity

Active Publication Date: 2016-04-20
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: 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
6) Drug susceptibility test: This method is time-consuming, cumbersome to operate, and relies on the isolation and culture of strains

Method used

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  • System for detecting multiple drug-resistant and quantitative genes of H.pylori as well as kits and applications of system
  • System for detecting multiple drug-resistant and quantitative genes of H.pylori as well as kits and applications of system
  • System for detecting multiple drug-resistant and quantitative genes of H.pylori as well as kits and applications of system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[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 (TaqDNA Polymerase), positive and negative controls. 2mmol / L dNTPs and fluorescent universal labeling mixture are mixed together into a 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 mixture includes a forward primer for the 16SrRNA gene, a reverse primer for the 16SrRNA gene, a forward primer for the cagA gene, a reverse primer for the cagA gene, a forward primer for the vacA-s1 or vacA-s2 gene, Reverse primer for vacA-s1 or vacA-s2 gene, forward primer for vacA-m1 gene, reverse ...

Embodiment 2

[0116] The Helicobacter pylori detection kit of this embodiment is the same as the rest of Example 1, except that the primer mixture only includes the forward primer for the 16SrRNA gene, the reverse primer for the 16SrRNA gene, and the forward primer for the 23SrRNA gene. Primers, reverse primer corresponding to base A at the 2143 site of the 23SrRNA gene, reverse primer corresponding to base G at the 2143 site of the 23SrRNA gene, forward primer for the rdxA gene, and base C corresponding to the 148 site of the rdxA gene The reverse primer corresponding to the 148th position of the rdxA gene is base T, the forward primer for the pbp1A gene, the reverse primer corresponding to the 1777th position of the pbp1A gene is the base A, and the corresponding pbp1A gene 1777th position is Reverse primer for base G, forward primer for gyrA gene, reverse primer corresponding to base C or T at site 261 of gyrA gene, reverse primer corresponding to base G or A at site 261 of gyrA gene, Fo...

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Abstract

The invention relates to a system for detecting multiple drug-resistant and quantitative genes of H.pylori as well as kits and applications of the system. The system for detecting multiple drug-resistant and quantitative genes of H.pylori comprises multiple pairs of primers, eachforstrain identification genes (16S rRNA), drug-resistant genes (polymorphism of a 2143rd locus of 23S rRNA, a 148th locus of rdxA, a 1777th locus of pbp1A and a 261st locus ofgyrA), and quantitative analysis genes ureC and beta-globin. The system for detecting multiple drug-resistant and quantitative genes and the kits of the system can directly perform synchronous detection and analysis on the strain identification, drug resistance and quantification 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 guide the accurate diagnosis and individualized medication of H.pylori infection and the observation of curative effects.

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 mainly resides in 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. Epid...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6851C12Q1/689C12Q2600/16
Inventor 张艳梅赵虎胡彬婕王诗雯赵付菊陈飞缪应新徐玲丽吴勇南丽孔咪咪姜文荣张景皓
Owner HUADONG HOSPITAL
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