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Kit for detecting polymorphism of hypertension medication related genes

A gene polymorphism and kit technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., to achieve the effects of low cost, simple and feasible interpretation method, and simple operation

Pending Publication Date: 2020-05-05
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Sensitivity only reaches 0.1ng / μL

Method used

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  • Kit for detecting polymorphism of hypertension medication related genes
  • Kit for detecting polymorphism of hypertension medication related genes
  • Kit for detecting polymorphism of hypertension medication related genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0104] Embodiment 1: the composition of kit

[0105] The composition of kit of the present invention is as follows:

[0106] Table 2 Composition of the kit

[0107]

[0108] Table 3 Primers and Probes

[0109]

[0110]

Embodiment 2

[0111] Embodiment 2: Extraction of sample genomic DNA

[0112] In this example, human whole blood / peripheral blood samples were collected, and genomic DNA was extracted therefrom.

[0113] Genomic DNA was extracted from EDTA anticoagulated plasma and used as a template for PCR detection. Use pretreatment reagents and nucleic acid extraction and purification reagents (magnetic bead method) from Zhengzhou Antu Bioengineering Co., Ltd., and follow the instructions. The details are as follows:

[0114] 1. Sample pretreatment:

[0115] 1) Pretreatment of frozen-thawed whole blood with red blood cell lysate:

[0116] ①Take 200 μL clinical frozen-thawed or hemolyzed whole blood sample, add 600 μL red blood cell lysate, and gently invert and mix 10 times. After the red blood cell is lysed, the solution should be clear and transparent. 12000rpm / min, centrifuge for 3min, discard the supernatant;

[0117] ②Add 600μL 0.2mg / ml proteinase K, incubate at room temperature 20-25℃ for 5min,...

Embodiment 3

[0129] Embodiment 3: the detection of sample genomic DNA

[0130] In this example, the combination of primers and probes listed in Example 1 is used to amplify the genomic DNA sample extracted in Example 2. The detection steps are as follows:

[0131] ① Build a reaction system

[0132] Calculate the number of samples required, increase the amount of reagents in proportion to the amount in the table below, and only do one copy of negative and positive quality control.

[0133] Table 4 wild type reaction system W1

[0134]

[0135] Table 5 mutant reaction system M1

[0136]

[0137]

[0138] Table 6 Wild type reaction system W2

[0139]

[0140] Table 7 Mutant reaction system M2

[0141] PCR reaction solution (M2) PCR reaction solution (E) sample DNA positive control negative control 20 μL 10μL 50μL - - 20 μL 10μL - 50μL - 20 μL 10μL - - 50μL

[0142] ② On-board inspection

[0143] Directly detect on the f...

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PUM

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Abstract

The invention relates to the technical field of in vitro diagnosis, in particular to a kit for detecting polymorphism of human CYP2D6, CYP2C9, ADRB1, AGTR1, ACE genes by a multiple fluorescent PCR method. The kit is used to detect CYP2D6*10, CYP2C9*3, ADRB1 (1165G > C), AGTR1 (1166A > C), ACE (I / D) polymorphism sites. The primer and the probe have high sensitivity and high specificity, and can accurately detect genomic DNA as low as 0.1 ng / <mu>L. The kit is easy to operate, and can cooperate with an automated instrument for detection.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis, in particular to a kit for detecting human CYP2D6, CYP2C9, ADRB1, AGTR1 and ACE gene polymorphisms by a multiplex fluorescent PCR method. Background technique [0002] Individual differences in hypertension drugs are also very common in clinical practice. It is reported that about 20%-50% of hypertensive patients fail to achieve good blood pressure control after receiving drug treatment. The main reason for this phenomenon is also the genetic variation of drug-related drug-metabolizing enzymes and receptors. At present, the research on genetic variation of drug-metabolizing enzymes and receptors in hypertension mainly focuses on CYP2C9*3, ADRBI (1165G>C) , AGTRI (166A>C), CYP2D6*10, ACE (I / D) and other genes, the mutation of these genes is the root cause of individual differences in drug response in hypertensive patients. [0003] CYP2C9 (CytochromeP4502C9) is an important membe...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6858
CPCC12Q1/6883C12Q1/6858C12Q2600/106C12Q2600/156C12Q2600/16C12Q2600/166C12Q2531/113C12Q2537/143C12Q2535/137C12Q2561/101C12Q2521/531C12Q2545/101C12Q2545/113
Inventor 王双双王义聪薛方方王玮李振红付光宇杨增利苗拥军
Owner AUTOBIO DIAGNOSTICS CO LTD
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