Method for individual medication of valsartan through detection product

A technology for medication guidance and valsartan, applied in biochemical equipment and methods, microbiological determination/testing, DNA/RNA fragments, etc., can solve limitations and other problems

Inactive Publication Date: 2020-05-05
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 path

Method used

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  • Method for individual medication of valsartan through detection product
  • Method for individual medication of valsartan through detection product
  • Method for individual medication of valsartan through detection product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Example 1: Primer Design and Synthesis

[0084] 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.

[0085] The corresponding specific PCR primer core sequences (SEQ1a to SEQ3a) and specific extension primer core sequences (SEQ1b to SEQ3b) were designed for the polymorphic sites of rs1799983, rs1042713, and rs1801253 related to the discrimination of drug types. Three pairs of PCR primers and three extension primers constitute three independent reaction systems: SEQ1a / b constitutes the first reaction system, and SEQ2a / b to SEQ3a / b constitute the second reaction system. In these 2 independent reaction systems, SEQ1a, SEQ2a to SEQ3a respectively participate in 2 independent multiplex PCR reactions, and SEQ1b, SEQ2b to SEQ3b respectively participate in subsequent 2 independent single ...

Embodiment 2

[0087] Embodiment 2: sample DNA extraction

[0088] 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

[0089] Embodiment three: biological experiment

[0090] Using ABI 9700 type PCR instrument, according to the instruction manual, check the 2 polymorphic sites for identifying the drug type.

[0091] The components used in the kit for PCR, PCR product purification and single base extension are:

[0092] 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

[0093] The concentration of each primer pair is 500nmol / L.

[0094] According to the manual, the specific operation method is as follows:

[0095] 1. PCR amplification

[0...

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Abstract

The invention provides an extension primer with different molecular weights by utilizing different single nucleotide polymorphism (SNP) sites, so that a plurality of sites are simultaneously detectedby time-of-flight mass spectrometry (MALDI-TOF MS), and finally a method for judging valsartan individualized medication by mass spectrometry of a detection product is obtained. The method comprises the following steps: respectively designing a multiple amplification primer and an extension primer according to a plurality of target SNP sites to be detected; preparing a multiple amplification primer reaction system and an extension primer reaction system; in the reaction system, simultaneously carrying out amplification and a single-base extension reaction on the plurality of target SNP sites by using a plurality of sets of primers; and carrying out time-of-flight mass spectrometry analysis on a product after the single-base extension reaction, identifying genotypes of SNP related to metabolism of different drugs, and guiding individualized medication of the hypertension-lowering drug valsartan. The method has the advantages of low cost, short time consumption, simple and convenient result analysis and extremely wide 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 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 target oligonucleotide fragments in the process of mass spectrometry typing, and simultaneously detects multiple target SNP sites to guide the antihypertensive drug valsa. Tan's personalized medicine. 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...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/106C12Q2600/156C12Q2600/16
Inventor 马庆伟钟逾刘昕超张海燕
Owner BIOYONG TECH
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