Primer group for detecting gene polymorphism sites relevant to drug metabolic enzyme activity, application thereof, product applying primer group, and detection method

A technology of primer sets and metabolic enzymes, applied in the field of genetic engineering, can solve the problems of simultaneous detection of multiple sites, time-consuming and labor-intensive problems, and achieve the effects of wide clinical application range, accurate detection, and simple operation

Inactive Publication Date: 2018-03-27
山东龙辰生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The first object of the present invention is to provide a primer set for detecting gene polymorphic sites related to the activity of drug-metabolizing enzymes, so as to alleviate the time-consuming and labor-intensive problems in the prior art that multiple sites cannot be detected simultaneously. technical issues

Method used

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  • Primer group for detecting gene polymorphism sites relevant to drug metabolic enzyme activity, application thereof, product applying primer group, and detection method
  • Primer group for detecting gene polymorphism sites relevant to drug metabolic enzyme activity, application thereof, product applying primer group, and detection method
  • Primer group for detecting gene polymorphism sites relevant to drug metabolic enzyme activity, application thereof, product applying primer group, and detection method

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Experimental program
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Effect test

Embodiment 1

[0069] The processing of embodiment 1 sample

[0070] In the Inpatient Department of Cardiology Department of Linyi People's Hospital, the clinical medical records of patients who routinely took clopidogrel after percutaneous coronary stenting and acute coronary syndrome, as well as the ADP-induced platelets detected by thromboelastography (TEG) Inhibition rate, according to the selection criteria, 100 patients were selected, and about 5mL of peripheral venous blood was collected, white blood cells were separated and DNA was extracted.

[0071] DNA extraction was completed in the Central Laboratory of Linyi People's Hospital.

[0072] DNA Extraction Kit: Human-derived DNA Quantitative Kit (Applied Biosystems, USA) is used for DNA quantitative detection, including the following reagents and materials: Buffer AL, Buffer AW1, Buffer AW2, AE, proteinase K, spin column, collection tube .

[0073] DNA extraction method (spin column method):

[0074] (1) Take a new sterilized cent...

Embodiment 2

[0086] The detection of embodiment 2 samples

[0087] CYP2C19*2, *3, ABCB1, PON1 were genotyped by real-time quantitative PCR technology. The sequences of primers and probes used in the experiments are provided by the present invention.

[0088] (1) Configuration of PCR reaction system

[0089] Add dUTP, Buffer, MgCl in the kit to each reaction well 2 , HsTaq, template DNA, internal reference primers and internal reference probe PCR reaction system is as follows:

[0090] The wild-type PCR amplification system is:

[0091] Reagent

[0092] The mutant PCR amplification group system is:

[0093]

[0094]

[0095] The PCR reaction procedure is:

[0096]

[0097] Result after reading:

[0098] genotype

[0099] In summary, the primer set provided by the present invention for detecting gene polymorphic sites related to drug-metabolizing enzyme activity can simultaneously detect different kinds of gene polymorphic sites related to drug-metabolizing e...

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Abstract

The invention provides a primer group for detecting gene polymorphism sites relevant to drug metabolic enzyme activity, application thereof, a product applying the primer group, and a detection method, which relate to the field of a genetic engineering technology. The primer group for detecting the gene polymorphism sites relevant to drug metabolic enzyme activity provided by the invention comprises a primer pair and a probe for detecting a CYP2C19 gene CYP2C19*2 site, a CYP2C19*3 site, an ABCB1 gene 3435C site, a PON1 gene rs854560 site, can be used for detecting different species of gene polymorphism sites relevant to drug metabolic enzyme activity at the same time, and is comprehensive in detecting and wide in application, and a detection result can be used for clinic guiding pharmacy.A reagent and a kit for detecting a medicine poor metabolizer provided by the invention comprise the primer group or a reagent provided by the invention, and are low in cost, high in efficiency, capable of quickly and accurately detecting the medicine poor metabolizer, high in sensitivity and high in specificity.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a primer set for detecting gene polymorphic sites related to drug metabolizing enzyme activity, its application, products applied thereto, and a detection method. Background technique [0002] The drug metabolism process is completed by a series of enzymatic reactions, and there are two types of enzymes involved: microsomal enzymes and non-microsomal enzymes. Microsomal enzymes mainly exist in the liver, lungs, kidneys, small intestine, placenta, skin and other parts. The activity of microsomal enzymes in the liver is the highest, mainly catalyzing the metabolism of exogenous substances such as drugs, so they are also called drug metabolizing enzymes , referred to as drug enzyme. Drug-metabolizing enzymes play an important catalytic role in the metabolism of drugs. After drugs enter the human body, on the one hand, they affect the body to produce pharmacological eff...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/106C12Q2600/166C12Q2600/16C12Q2600/156
Inventor 张守村孙相鑫薛澄
Owner 山东龙辰生物技术有限公司
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