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Medication guidance and gene detection kit for antihypertensive drug irbesartan

A high blood pressure and drug technology, applied in the field of detection of multiple PCR single base extension products, can solve limitations and other problems

Inactive Publication Date: 2020-05-26
BIOYONG TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the method and kit provided by this invention patent can only detect polymorphic sites on the homocysteine ​​metabolism pathway, and cannot detect polymorphic sites not involved in homocysteine ​​metabolism , has limitations in clinical medication guidance

Method used

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  • Medication guidance and gene detection kit for antihypertensive drug irbesartan
  • Medication guidance and gene detection kit for antihypertensive drug irbesartan
  • Medication guidance and gene detection kit for antihypertensive drug irbesartan

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Example 1: Primer Design and Synthesis

[0082] The surrounding sequences of the target SNP of this kit are queried in the db_SNP (build 132) and Hapmap (Rel 28, Phase II + III, Aug 10) databases, and these sequences are used to design multiplex PCR primers and single base extension primers.

[0083] The corresponding specific PCR primer core sequence (SEQ 1a) and specific extension primer core sequence (SEQ 1b) were designed for the polymorphic site of rs1367117 related to the discrimination of the drug type. One pair of PCR primers and one extension primer constitute an independent reaction system: SEQ 1a / b. In this independent reaction system, SEQ 1a participates in an independent multiplex PCR reaction, and SEQ 1b participates in a subsequent independent single-base extension reaction.

[0084] Related primers were synthesized at Sangon Bioengineering (Shanghai) Co., Ltd.

Embodiment 2

[0085] Embodiment 2: sample DNA extraction

[0086] A total of 10 DNA samples were collected from ordinary Chinese people, marked as A1-A10. Among them, sample collection, DNA extraction, etc. were collected in accordance with the requirements of the instructions, and human venous blood was collected with EDTA anticoagulant tubes. According to the instructions, the collected blood should not be stored at 2-8°C for more than one week, and at -20°C for no more than one month, and can be transported in a curling box with ice or a foam box with ice. It is recommended to use fresh blood as much as possible. Genomic DNA extraction. Since this kit does not provide human genomic DNA extraction reagents, a commercial nucleic acid extraction kit (such as QIAGEN’s DNeasy Blood and Tissuekit) was used to extract human genomic DNA from 200 μL of whole blood of each patient, and the DNA was extracted using a NanoDrop 2000 ( Thermo Company) quantified, and standardized to 30ng / μL (respecti...

Embodiment 3

[0087] Embodiment three: biological experiment

[0088] Using ABI 9700 type PCR instrument, according to the instruction manual, check the 2 polymorphic sites for identifying the drug type.

[0089] The components used in the kit for PCR, PCR product purification and single base extension are:

[0090]

[0091]

[0092] The concentration of each primer pair is 500nmol / L.

[0093] According to the manual, the specific operation method is as follows:

[0094] 1. PCR amplification

[0095] 1.1 In the PCR preparation area, prepare 200 μL PCR reaction tubes according to the number of samples to be tested (including positive quality control, negative control, and blank control), and mark the sample number on the tube;

[0096] 1.2 Take out the PCR primer mixture and PCR reaction solution from the kit, let them thaw naturally, vortex and oscillate to mix well, and centrifuge instantly to the bottom of the tube;

[0097] 1.3 According to the number of samples, take out the P...

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Abstract

The invention provides a medication method and a kit of an antihypertensive drug irbesartan. The method comprises the following steps: respectively designing multiple amplification primers and extension primers according to a plurality of to-be-detected target SNP loci; preparing a multiplex amplification primer reaction system and an extension primer reaction system; in a reaction system, using aplurality of primers for simultaneously and respectively carrying out amplification and single base extension reaction on a plurality of target SNP sites; carrying out flight time mass spectrometry analysis on a product obtained after the single base extension reaction, identifying genotypes of SNP related to different drug metabolism according to products of extension primers with different molecular weights represented by mass spectrometry peaks, and guiding medication of the antihypertensive drug irbesartan. The invention also provides the detection kit using the method. The SNP locus related to metabolism of the antihypertensive drug irbesartan can be detected, and the SNP locus has the advantages of being low in cost, free of probe synthesis, short in consumed time, simple and convenient in result analysis and extremely wide in application field.

Description

technical field [0001] The invention belongs to the field of molecular biology detection, and relates to a method for detecting multiple PCR single-base extension products by using a characteristic peak diagram of a mass spectrum and a product thereof. More specifically, the method uses different time-of-flight mass spectrometry characteristic peaks generated by different target oligonucleotide fragments in the process of mass spectrometry typing, and simultaneously detects multiple target SNP sites to guide the antihypertensive drug Erbe Sartan medication. Background technique [0002] Human genetic information is stored in the genome. In 2002, the Human Genome Project, an international cooperation project, was finally completed, drawing a fine map of the human genome structure and providing a reference sequence for related research. The human genome sequence is composed of 3 billion bases. A large number of studies have shown that >1% of the genetic differences in the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6883C12Q2600/106C12Q2600/156C12Q2600/16C12Q2531/113C12Q2537/143C12Q2533/101C12Q2565/627
Inventor 马庆伟钟逾刘昕超
Owner BIOYONG TECH
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