Multiple-gene helicobacter pylori detection system and kit and application thereof

A Helicobacter pylori and gene detection technology, applied in the biological field, can solve the problems of relying on isolated and cultured strains, cumbersome operations, and low throughput, so as to guide individualized medication and curative effect monitoring, avoid mutual interference, and compensate for low throughput. Effect

Active Publication Date: 2016-03-30
HUADONG HOSPITAL +1
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  • Abstract
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  • Claims
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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, cumber

Method used

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  • Multiple-gene helicobacter pylori detection system and kit and application thereof
  • Multiple-gene helicobacter pylori detection system and kit and application thereof
  • Multiple-gene helicobacter pylori detection system and kit and application thereof

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Embodiment

[0051] 1. Composition of the kit

[0052] The Helicobacter pylori detection kit in this embodiment includes: primer mixture, PCR buffer (10×PCRBuffer), MgCl 2 Solution, dNTPs, fluorescent universal label mix, hot-start DNA polymerase (Taq DNA polymerase), positive and negative controls. 2 mmol / L dNTPs and fluorescent universal label mixture are mixed together to form a reagent tube.

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

[0054] Positive controls are plasmid mixtures that include all gene targets of interest.

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

[0056] The primer mix includes forward primer for 16SrRNA gene, reverse primer for 16SrRNA gene, forward primer for cagA gene, reverse primer for cagA gene, forward primer for vacA-s1 or vacA-s2 gene, Reverse primer for vacA-s1 or vacA-s2 gene Forward primer for vacA-m1 gene, reverse primer for vacA-m1 gene, forward primer for vacA...

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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 multiplex 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 bacterium that resides primarily 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 extensive clinical attention. In 1994, the International Agency for Research on Cancer (IARC) has classified it as a human class I carcinogen, and it is the only bacterial pathogenic microorganism that has been classified as clearly carcinogenic to humans so far. Many reports suggest that Helicobacter pylori infection is related to coronary heart disease, rheumatoid disease, hepatobiliary disease, tuberculosis, pregnancy vomiting...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12Q1/06
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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