Medication guide of antihypertensive drug ramipril and gene detection test kit

A high blood pressure and drug technology, applied in the field of oligonucleotide products, can solve the problems of difficult to meet the needs of fast and accurate, cumbersome operation, limited detection objects, etc.

Inactive Publication Date: 2020-05-01
BIOYONG TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the detection method and kit provided by this invention patent only target the polymorphic sites of key enzyme genes on the homocysteine ​​metabolic pathway, and the detection objects are limited, and the operation is cumbersome, and it is difficult to meet the requirements of clinical fast and accurate needs

Method used

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  • Medication guide of antihypertensive drug ramipril and gene detection test kit
  • Medication guide of antihypertensive drug ramipril and gene detection test kit
  • Medication guide of antihypertensive drug ramipril and gene detection test kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Example 1: Primer Design and Synthesis

[0083] 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 multiple PCR primers and single base extension primers.

[0084] The corresponding specific PCR primer core sequences (SEQ1a to SEQ2a) and specific extension primer core sequences (SEQ1b to SEQ2b) were designed for the polymorphic sites of rs4359 and rs4344 related to the discrimination of drug types. 2 pairs of PCR primers and 2 extension primers constitute an independent reaction system: SEQ1a / b to SEQ2a / b. In this independently performed reaction system, SEQ1a to SEQ2a participate in an independent multiplex PCR reaction, and SEQ1b to SEQ2b participate in a subsequent independent single base extension reaction.

[0085] Related primers were synthesized at Sangon Bioengineering (Shanghai) Co., Ltd.

Embodiment 2

[0086] Embodiment 2: sample DNA extraction

[0087] 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

[0088] Embodiment three: biological experiment

[0089] Using ABI 9700 type PCR instrument, according to the instruction manual, check the 2 polymorphic sites for identifying the drug type.

[0090] The components used in the kit for PCR, PCR product purification and single base extension are:

[0091] serial number component name main ingredient Specification 1 PCR Primer Mix PCR primers 24μl / tube x1 tube 2 PCR reaction solution Taq enzyme, dNTP 72μl / tube x1 tube 3 Enzyme digestion reaction solution SAP enzyme 48μl / tube x1 tube 4 Extension Primer Mix extension primer 24μl / tube x1 tube 5 Extension reaction solution Single base elongase, ddNTP 24μl / tube x1 tube 6 positive control Human Genomic DNA (30ng / μL) 10μL / tube x1 tube

[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

[0...

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Abstract

The invention provides a method and a medication method of antihypertensive drug ramipril and a gene detection test kit, wherein extension primers with different molecular weights at different singlenucleotide polymorphism (SNP) sites are utilized, therefore a plurality of sites can be simultaneously detected by a matrix-assisted laser desorption-ionization time of flight mass spectrometry(MALDI-TOF MS). The kit can simultaneously detect two ramipril drug metabolism related SNP sites, has the advantages of low cost, no need of probe synthesis, short time consumption, simple and convenient result analysis and extremely wide application field, and can effectively reduce occurrence of adverse events related to ramipril use amount and prevention of thrombotic diseases. The ramipril injectioncan be used for auxiliary diagnosis and treatment of hypertension patients who need to take ramipril clinically.

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 mass spectrometry characteristic peaks and its products. The method can simultaneously detect multiple PCR single-base extensions in two multiplex PCR reactions Amplified oligonucleotide products. More specifically, the method uses different time-of-flight mass spectrometry characteristic peaks generated by different purpose oligonucleotide fragments in the process of mass spectrometry typing, and simultaneously detects multiple target SNP sites to guide the antihypertensive drug Remy Puli's 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 geno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/106C12Q2600/156C12Q2600/16
Inventor 马庆伟钟逾
Owner BIOYONG TECH
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