Kit for evaluating medicine of glonoin

Abstract

The invention discloses a kit for detecting the sensibility of an individual to glonoine medicines. The kit comprises specific primer pair for detecting SNP polymorphism genotype rs671 on ALDH2 gene, a specific fluorescent probe pair and a routine component used for fluorescent quantitative PCR detection. The sensibility of the individual to the glonoine medicines is analyzed and evaluated by detecting the genotype of SNP locus rs671on the ALDH2 gene.

Description

technical field

[0001] The invention relates to the fields of molecular biology and medicine, and more specifically, the invention relates to a kit for detecting the drug sensitivity of an individual to nitroglycerin. Background technique

[0002] Nitroglycerin (GTN, glyceryl trinitrate, nitroglycerin) has been used in the treatment of angina pectoris and heart failure for 130 years. Sublingual nitroglycerin tablets are the routine first-choice prescription for anti-angina pectoris acute attack of coronary heart disease, but its clinical effectiveness is often due to It varies from person to person. Some people saved their lives in time because of it, but after some people took it, the effect was not very obvious. In the past, the domestic and foreign medical circles have always believed that the poor curative effect of nitroglycerin on some patients is caused by its "drug resistance", but the research results prove that this is not the case. Recent studies have shown that...

Examples

Embodiment 1

[0023] Example 1. Use of detection kits

[0024] Step 1: Extraction of DNA template

[0025] Genomic DNA of oral epithelial cells was extracted by silica gel adsorption.

[0026] Step 2: Real-time quantitative PCR reaction

[0027] Use the detection kit to carry out an independent fluorescent quantitative PCR reaction. The reaction system has a total volume of 10 μl, including 2 μl of DNA template with a concentration of 20 ng / μl, 1 μl of 10 X fluorescent quantitative PCR reaction buffer, 0.1 μl of 25 mM dNTP mixture, 0.6 μl 25 mM MgCl 2 solution, 0.025 μl (5 units / μl) Taq DNA polymerase, 0.225 μl each of 20 μM sense primer and antisense primer, 0.25 μl each of 10 μM fluorescent probe with VIC and fluorescent probe with FAM, and 5.325 μl of deionized water.

[0028] The reaction was carried out on a PCR amplification instrument, and the reaction conditions were 50° C. for 2 minutes, 95° C. for 10 minutes, and 60 cycles of 95° C. for 30 seconds and 60° C. for 1 minute. Afte...

Embodiment 2

[0032] Example 2. Service to evaluate people for nitroglycerin medication

[0033] Step 1: DNA Extraction

[0034] The physicians in the laboratory department of the hospital instructed the subjects to use oral swabs to sample oral epithelial cells, and the silica gel adsorption method was used to extract DNA from oral epithelial cells.

[0035] Step 2: Genotyping Assays

[0036] Using the kit provided by the invention, perform fluorescence quantitative PCR detection on the SNP site rs671 on the ALDH2 gene of the subject's genomic DNA, and determine the genotype of the SNP site.

[0037] Step 3: Evaluation Analysis of Individual Nitroglycerin Medication

[0038] Through the analysis of the SNP genotype of the tested subjects, an evaluation and analysis report of individual nitroglycerin medication is issued. The report details the effectiveness of the tested subject's nitroglycerin medication, which can be used as a reference for physicians to adjust the patient's medicatio...

technical field

[0001] The invention relates to the fields of molecular biology and medicine, and more specifically, the invention relates to a kit for detecting the drug sensitivity of an individual to nitroglycerin. Background technique

[0002] Nitroglycerin (GTN, glyceryl trinitrate, nitroglycerin) has been used in the treatment of angina pectoris and heart failure for 130 years. Sublingual nitroglycerin tablets are the routine first-choice prescription for anti-angina pectoris acute attack of coronary heart disease, but its clinical effectiveness is often due to It varies from person to person. Some people saved their lives in time because of it, but after some people took it, the effect was not very obvious. In the past, the domestic and foreign medical circles have always believed that the poor curative effect of nitroglycerin on some patients is caused by its "drug resistance", but the research results prove that this is not the case. Recent studies have shown that...

Examples

Embodiment 1

[0023] Example 1. Use of detection kits

[0024] Step 1: Extraction of DNA template

[0025] Genomic DNA of oral epithelial cells was extracted by silica gel adsorption.

[0026] Step 2: Real-time quantitative PCR reaction

[0027] Use the detection kit to carry out an independent fluorescent quantitative PCR reaction. The reaction system has a total volume of 10 μl, including 2 μl of DNA template with a concentration of 20 ng / μl, 1 μl of 10 X fluorescent quantitative PCR reaction buffer, 0.1 μl of 25 mM dNTP mixture, 0.6 μl 25 mM MgCl 2 solution, 0.025 μl (5 units / μl) Taq DNA polymerase, 0.225 μl each of 20 μM sense primer and antisense primer, 0.25 μl each of 10 μM fluorescent probe with VIC and fluorescent probe with FAM, and 5.325 μl of deionized water.

[0028] The reaction was carried out on a PCR amplification instrument, and the reaction conditions were 50° C. for 2 minutes, 95° C. for 10 minutes, and 60 cycles of 95° C. for 30 seconds and 60° C. for 1 minute. Afte...

Embodiment 2

[0032] Example 2. Service to evaluate people for nitroglycerin medication

[0033] Step 1: DNA Extraction

[0034] The physicians in the laboratory department of the hospital instructed the subjects to use oral swabs to sample oral epithelial cells, and the silica gel adsorption method was used to extract DNA from oral epithelial cells.

[0035] Step 2: Genotyping Assays

[0036] Using the kit provided by the invention, perform fluorescence quantitative PCR detection on the SNP site rs671 on the ALDH2 gene of the subject's genomic DNA, and determine the genotype of the SNP site.

[0037] Step 3: Evaluation Analysis of Individual Nitroglycerin Medication

[0038] Through the analysis of the SNP genotype of the tested subjects, an evaluation and analysis report of individual nitroglycerin medication is issued. The report details the effectiveness of the tested subject's nitroglycerin medication, which can be used as a reference for physicians to adjust the patient's medicatio...