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Mass spectrometry method for identifying personalized medication of trandolapril through product detection

A technology of trandolapril and mass spectrometry, applied in the field of oligonucleotide products, can solve problems such as difficulty in meeting fast and accurate requirements, limitations of detection objects, and cumbersome operations

Inactive Publication Date: 2020-05-29
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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  • Mass spectrometry method for identifying personalized medication of trandolapril through product detection
  • Mass spectrometry method for identifying personalized medication of trandolapril through product detection
  • Mass spectrometry method for identifying personalized medication of trandolapril through product detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Example 1: Primer Design and Synthesis

[0086] 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.

[0087] The corresponding specific PCR primer core sequences (SEQ1a to SEQ5a) and specific extension primer core sequences (SEQ1b to SEQ5b) were designed for five polymorphic sites related to the discrimination of drug types, including rs699, rs2016848, rs11209716, rs4742610, and rs8012552. 5 pairs of PCR primers and 5 extension primers form 3 independent reaction systems: SEQ1a / b to SEQ2a / b form the first reaction system, SEQ3a / b to SEQ4a / b form the second reaction system, and SEQ5a / b forms the The third reaction system. In these 3 independent reaction systems, SEQ1a to SEQ2a, SEQ3a to SEQ4a, and SEQ5a respectively participate in 3 independent multiplex PCR reactions, and SEQ1b to...

Embodiment 2

[0089] Embodiment 2: sample DNA extraction

[0090] 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

[0091] Embodiment three: biological experiment

[0092] Using ABI 9700 type PCR instrument, according to the instructions, check the 5 polymorphic sites that are used to identify the drug type.

[0093] The components used in the kit for PCR, PCR product purification and single base extension are:

[0094] 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

[0095] The concentration of each primer pair is 500nmol / L.

[0096] According to the manual, the specific operation method is as follows:

[0097] 1. PCR amplification ...

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Abstract

The invention provides a mass spectrometry method for identifying personalized medication of trandolapril through product detection. The method includes the steps: designing multiple-amplification primers and extension primers separately according to multiple to-be-detected target SNP sites, preparing a multiple amplification primer reaction system and an extension primer reaction system, performing amplification and single-base extension reactions at the target SNP sites simultaneously through multiple sets of primers in the reaction systems, performing time-of-flight mass spectrometry analysis of products obtained after the single-base extension reactions so as to identify the genotypes of SNP related to metabolism of different drugs, and guiding personalized medication of the antihypertensive drug trandolapril. Five SNP sites related to metabolism of the trandolapril drugs can be detected simultaneously, and the method has the advantages of low cost, no needs for synthetic probes, short time consumption, simple and convenient result analysis and extremely wide application fields, and can be applied to auxiliary diagnosis and treatment of hypertensive patients who need to take trandolapril 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 three multiplex PCR reactions Amplified oligonucleotide products. More specifically, the method utilizes 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 groups. 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 h...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6858
CPCC12Q1/6858C12Q1/6883C12Q2600/106C12Q2600/156C12Q2600/16C12Q2531/113C12Q2537/143C12Q2533/101C12Q2565/627
Inventor 马庆伟钟逾刘昕超张海燕
Owner BIOYONG TECH
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