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A gene chip for detection of individualized treatment of Helicobacter pylori infection and its application

A technology for detecting gene chips and Helicobacter pylori, which is applied in the field of medical inspection and gene chips, individualized treatment of Helicobacter pylori infection and detection of gene chips

Active Publication Date: 2016-09-28
INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, domestic practical chip research has just started. Up to now, there is no rapid gene detection chip for individualized treatment of Helicobacter pylori infection in my country.

Method used

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  • A gene chip for detection of individualized treatment of Helicobacter pylori infection and its application
  • A gene chip for detection of individualized treatment of Helicobacter pylori infection and its application
  • A gene chip for detection of individualized treatment of Helicobacter pylori infection and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Preparation of a gene chip for detection of individualized treatment of Helicobacter pylori infection

[0046] 1. Preparation of specific primers

[0047] Select the clarithromycin-resistant mutation sites A2142G and A2143G in the Helicobacter pylori 23s gene; the quinolone-resistant catastrophe sites Ash87 and Asp91 in the Helicobacter pylori gyrA gene; 2C19*2(G6981A) and 2C19*3(G636A) as detection targets

[0048] Specific primers were designed according to the standard (see Table 1). All downstream primers were 5'-end labeled with biotin.

[0049] Table 1 Specific primers and their corresponding amplification target gene types

[0050]

[0051] 2. Design and screen probes for human proton pump inhibitor metabolism-related sites and Helicobacter pylori drug-resistant sites

[0052] Wild-type and mutant SNP typing probes were designed for A2142G, A2143G, N87K, D91G / Y / N, G681A, and G636A, respectively (as shown in Table 2). Simultaneously design matri...

Embodiment 2

[0071] Example 2: Determination of Gene Chip Positive Judgment Criteria

[0072] All result judgments are based on the signal value calculated by the software, which reflects the gray value of the probe. The chip matrix control points, negative quality control points, and the positive value interpretation standards of the human genome and Helicobacter pylori drug resistance site probes are all calculated according to statistical methods. For the two probes of CYP450-2C19*2 (2C19*2-W and 2C19*2M), when the average gray value of a certain probe is twice or more than that of the other, it is judged that the probe is positive ; When the ratio of the average gray value of the two is less than 2, it is judged that the locus is heterozygous. The interpretation criteria of the two probes (2C19*3-W and 2C19*3M) of CYP450-2C19*3 are the same as those of CYP450-2C192. For the 4 probes (42W-43W, 42M-43W, 42W-43M and 42M-43M) of the clarithromycin-resistant site, when the average gray va...

Embodiment 3

[0073] Example 3: Specificity evaluation of gene chip detection for individualized treatment of Helicobacter pylori infection

[0074] Specificity is the most important assessment index of a diagnostic method. The present invention uses optimized systems and conditions to detect various strains and human genomes with common mutations. The chip detection results are shown in the attached figure 2 . It can be seen from the figure that the present invention can correctly distinguish bacterial strains with common mutations and human genome, and the specificity is good.

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Abstract

The invention relates to a biological engineering detection product and an application of the gene chip, and in particular relates to a detection gene chip for helicobacter pylori infection individualized treatment. The chip is capable of simultaneously detecting polymorphisms of helicobacter pylori clarithromycin medicine-resistant mutation sites A2142G and A2143G, carbostyril medicine-resistant mutation sites Asn87(N87K) and Asp91(D91G / Y / N), and sites 2C19*2(G6981A) and 2C19*3(G636A) related to metabolism of a proton pump inhibitor on human cytochrome enzyme CYP450. The chip detection method disclosed by the invention is rapid and accurate and is capable of identifying helicobacter pylori etiology, carrying out medicine-resistant analysis of clarithromycin and carbostyril, and analyzing metabolic individual differences of the proton pump inhibitor, so that the detection gene chip is used for guiding individualized administration of helicobacter pylori infection triplex process treatment.

Description

technical field [0001] The invention relates to a gene chip for individualized treatment and detection of Helicobacter pylori infection and its application, and relates to the technical field of medical examination and gene chip technology. It is a detection-type gene chip for individualized treatment of Helicobacter pylori infection. It can simultaneously detect CYP4502C19*2 and CYP4502C19*3 polymorphisms in the human genome and the resistance sites of clarithromycin and quinolones in Helicobacter pylori. Background technique [0002] Helicobacter pylori (H. pylori, HP) is a class I carcinogen identified by the WHO International Agency for Research on Cancer. The HP infection rate in my country is 40%-90%, with an average of 59%, which belongs to the hardest-hit area of ​​HP-related diseases. HP infection is an important cause of chronic gastritis and the occurrence and recurrence of peptic ulcer, and is closely related to the occurrence and development of low-grade gastri...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04C40B40/06C40B50/06
Inventor 王升启姚雪刘琪琦陈苏红张敏丽朱坤
Owner INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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