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Nucleic acid composition for CYP2C19 gene typing detection as well as detection kit and application

A CYP2C19 and detection kit technology, applied in the field of medical testing, can solve the problems of high cost of reagent detection, false positives, false negatives, etc., and achieve the effects of convenient detection reaction, rapid genotyping, great use value and social value

Inactive Publication Date: 2018-03-13
SUREXAM BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have more or less defects such as high cost of reagent detection, complicated operation, easy to produce pollution, and certain false negatives and false positives in the promotion of these methods.

Method used

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  • Nucleic acid composition for CYP2C19 gene typing detection as well as detection kit and application
  • Nucleic acid composition for CYP2C19 gene typing detection as well as detection kit and application
  • Nucleic acid composition for CYP2C19 gene typing detection as well as detection kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] This embodiment provides a nucleic acid combination for CYP2C19 genotyping detection. The nucleic acid combination includes a detection primer combination and a detection probe combination.

[0056] The detection primer combination comprises the first primer pair, the second primer pair, the third primer pair and the fourth primer pair; the base sequences of the first primer pair are shown in SEQ ID No.1-2 respectively, and the base sequences of the second primer pair The base sequences are respectively shown in SEQ ID No.3-4, the base sequences of the third primer pair are respectively shown in SEQ ID No.5-6, and the base sequences of the fourth primer pair are respectively shown in SEQ ID No.7-8 shown.

[0057] The combination of detection probes includes a first wild type detection probe, a first mutant detection probe, a second wild type detection probe, a second mutant detection probe, a third wild type detection probe, a third mutant detection probe Probe, the fo...

Embodiment 2

[0064] This embodiment provides a detection kit for CYP2C19 genotyping detection. The kit contains the nucleic acid combination provided in Embodiment 1. The detection primers include a pair of primers. The base sequences of the pair of primers are respectively shown in SEQ ID No.1-2. Show.

[0065] The detection kit for CYP2C19 genotyping detection also includes at least one of PCR reaction buffer, Taq DNA polymerase, and dNTPs.

[0066] The PCR reaction buffer is a 10× reaction buffer consisting of 500mM KCl, 25mM MgCl 2 , 100mM Tris and double distilled deionized water according to the ratio of 1:1:1:2 after autoclaving.

[0067] The detection kit for CYP2C19 genotyping detection also includes genome extraction reagents.

[0068] The specific usage method of the detection kit for CYP2C19 genotyping detection provided in this example is as follows:

[0069] Sample genome extraction, the specific steps are as follows:

[0070] Sample preparation: Take the blood sample fro...

experiment example 1

[0089] This experimental example provides detection of the specificity of the nucleic acid combination in the kit provided in Example 2.

[0090] The first primer pair corresponds to the CYP2C19*2 gene, the mutation site is G681A; the second primer pair corresponds to the CYP2C19*3 gene, the mutation site G636A; the third primer pair corresponds to the CYP2C19*3 gene, the mutation site C-806T and the fourth primer All pairs correspond to the CYP2C19*17 gene mutation site C-3402T.

[0091] The wild-type plasmid, mutant plasmid and mixed plasmid corresponding to the four sites were respectively used as templates, and the detection kit provided in Example 2 was used for detection. Refer to Example 2 for the PCR reaction system and reaction procedure.

[0092] CYP2C19*2 gene, the experimental results of mutation site G681A are as follows Figure 1 to Figure 3 shown; figure 1 is the experimental result of the wild-type plasmid, figure 2 The experimental results of mutant plas...

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Abstract

The invention provides a nucleic acid composition for CYP2C19 gene typing detection as well as a detection kit and application, and belongs to the field of medical examination. The nucleic acid composition for CYP2C19 gene typing detection can accurately and specially participate in reaction, and can simply and quickly complete typing experimental detection. The nucleic acid composition for CYP2C19 gene typing detecting is applied to the detection kit for CYP2C19 gene typing detecting; and the specially manufactured detection kit can perform detection reaction more conveniently, can quickly perform gene typing, and has a relatively great using value and a relatively great social value.

Description

technical field [0001] The invention relates to the field of medical testing, in particular to a nucleic acid combination, a detection kit and an application for CYP2C19 genotyping detection. Background technique [0002] Clopidogrel (Plavix), as an oral antiplatelet drug, is widely used in the prevention and treatment of myocardial infarction, ischemic cerebral thrombosis, thromboembolism, atherosclerosis and vasculitis obliterans and other diseases. It is an ineffective prodrug. After oral administration, about 85% of clopidogrel is hydrolyzed by esterase, and only 15% of the drug is converted into an active ingredient by the cytochrome P450 enzyme system of the liver (cytochromeP450, CYP) to play a role. CYP2C19 is a very important drug-metabolizing enzyme, which mainly exists in liver microsomes. Many endogenous substrates, environmental pollutants and about 2% of clinical drugs are catalyzed by it. The FDA has added a black-box warning to the packaging of Plavix (clopi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/106C12Q2600/156C12Q2600/166
Inventor 陈海霞许嘉森吴诗扬
Owner SUREXAM BIO TECH
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