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Gene detection kit for medicine application instruction of hypertension reduction medicine telmisartan

A technology for telmisartan and high blood pressure, applied in the field of oligonucleotide products, can solve limitations and other problems

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 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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  • Gene detection kit for medicine application instruction of hypertension reduction medicine telmisartan
  • Gene detection kit for medicine application instruction of hypertension reduction medicine telmisartan
  • Gene detection kit for medicine application instruction of hypertension reduction medicine telmisartan

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Example 1: Primer Design and Synthesis

[0081]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.

[0082] The corresponding specific PCR primer core sequence (SEQ1a) and specific extension primer core sequence (SEQ1b) were designed for the polymorphic site of rs5443 related to the discrimination of drug type. One pair of PCR primers and one extension primer constitute an independent reaction system: SEQ1a / b. In this independently performed reaction system, SEQ1a participates in an independent multiplex PCR reaction, and SEQ1b participates in a subsequent independent single-base extension reaction.

[0083] Related primers were synthesized at Sangon Bioengineering (Shanghai) Co., Ltd.

Embodiment 2

[0084] Embodiment 2: sample DNA extraction

[0085] 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

[0086] Embodiment three: biological experiment

[0087] Use ABI 9700 type PCR instrument, according to the instruction manual to test the polymorphic sites and distinguish the drug type.

[0088] The components used in the kit for PCR, PCR product purification and single base extension are:

[0089] 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

[0090] The concentration of each primer pair is 500nmol / L.

[0091] According to the manual, the specific operation method is as follows:

[0092] 1. PCR amplification

[0093...

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Abstract

The invention provides a medicine using method and a kit of hypertension reduction medicine telmisartan. The method comprises the following steps: separately designing multiple amplification primers and extension primers according to a plurality of target SNP locus to be detected; preparing multiple amplification primer reaction systems and extension primer reaction systems; simultaneously performing amplification and single base extension reaction on the plurality of target SNP locus by using multiple sets of primers in the reaction systems; performing flight time mass spectrometry analysis on the product subjected to the single base extension reaction, verifying gene types of metabolism-related SNP of different medicines, and guiding using of the hypertension reduction medicine telmisartan. Meanwhile, the invention provides the detection kit using the method. According to the medicine using method and the kit disclosed by the invention, SNP locus related with the metabolism of the hypertension reduction medicine telmisartan can be detected, and the kit has the advantages of being low in cost, having no need of synthetic probe, being short in time consumption, being simple and convenient in result analysis, and being extremely wide in application field, and can be used for aided diagnosis and treatment of hypertension patients who clinically need oral administration of the telmisartan.

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 one multiplex PCR reaction 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 temami 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 geno...

Claims

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Application Information

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