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Method for detecting acute angina pectoris treating effect of nitroglycerin

A kit and in vitro detection technology, applied in the fields of molecular biology and medicine, can solve the problems of nitroglycerin treatment failure and unproven

Inactive Publication Date: 2006-03-22
CHINESE NAT HUMAN GENOME CENT AT SHANGHAI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, to date, the true cause of the failure of nitroglycerin therapy has not been identified

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] 1.1 Research object

[0064] Selected 80 Chinese Han patients with a definite diagnosis of coronary heart disease. They all took nitroglycerin as medicine for angina attack. According to the effect of sublingual nitroglycerin administration, these patients can be divided into an effective group within 10 minutes, and an ineffective group without a significant effect within 10 minutes. There were 59 patients in the effective group, including 47 males and 12 females. There were 21 patients in the invalid group, including 13 males and 8 females. For these patients, in terms of gender relationship, the effect of drug administration has no significant correlation with the patient's gender (p=0.107).

[0065] 1.2 Experimental methods and results

[0066] 1.2.1 DNA extraction

[0067] DNA was extracted from human blood by the conventional phenol-chloroform method, and the concentration was corrected to 20ng / ul for conventional PCR amplification.

[0068] 1.2.2 Design of ...

Embodiment 2

[0081] Activity determination of wild-type and mutant acetaldehyde dehydrogenase 2

[0082] From individuals heterozygous for the aldehyde dehydrogenase 2 genotype, the reverse transcription DNA (cDNA) of two different genotypes of the wild type and the 1951 G→A mutant type was obtained by reverse transcription, and sequenced. The resulting sequences were verified. Subsequently, a nested PCR (nest-PCR) method was used to amplify the target sequence and introduce restriction sites. Then, the amplified sequence was recombined into a plasmid (recombinantplasmid), and the plasmid was transfected into insect cells sf9 (purchased from Invitrogen). The sequences recombined into the plasmid were also verified by sequencing, which were SEQ ID NO: 5 (wild type, G at position 1510) and SEQ ID NO: 6 (mutant type, A at position 1510). The expression of the aldehyde dehydrogenase 2 gene from the plasmids transfected into the cells was verified using the conventional immunoblotting method....

Embodiment 3

[0090] Effectiveness prediction kit for nitroglycerin in the treatment of acute angina pectoris

[0091] As described in Example 1, the mutation at position 1951 G→A in SEQ ID NO: 1 is closely related to the efficacy of nitroglycerin in treating acute angina pectoris. Therefore, ALDH2 gene-specific primers can be designed based on this mutation and then amplified using the DNA of the individual to be tested as a template for prediction.

[0092] name

sequence

concentration

sense primer

SEQ ID NO: 3

100pmol

antisense primer

SEQ ID NO: 4

100pmol

PCR reaction solution

Containing Taq Enzyme dNTP Magnesium Ion PCR Reaction Buffer

[0093] Take 3ml of blood from the male patient to be tested, and use a conventional method (or use a specific kit) to extract DNA from the blood. Dilute the PCR primers in the Prediction Kit for Curative Effect of Nitroglycerin Treatment of Acute Angina Pectoris to 1 μmol / μl, and use the e...

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PUM

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Abstract

The method of predicting the effectiveness of nitroglycerin in treating acute angina pectoris includes detecting the individual acetaldehyde dehydrogenase 2 genes ALDH2, transcript and / or protein and the variation to the normal case, with the variation showing the lower effectiveness of nitroglycerin in treating acute angina pectoris compared with common acute angina pectoris herd. The present invention also discloses corresponding reagent kit.

Description

technical field [0001] The present invention relates to the fields of molecular biology and medicine. More specifically, it relates to the correlation between single nucleotide polymorphism (singlenucleotide polymorphism, SNP) of acetaldehyde dehydrogenase 2 gene (acetaldehyde dehydrogenase 2, ALDH2) and the curative effect of nitroglycerin in treating acute angina pectoris. The present invention also relates to methods and kits for detecting these SNPs. Background technique [0002] Nitroglycerin (NTG) has been widely used to treat acute angina since its first clinical application in 1876. Sublingual administration of nitroglycerin has become the standard treatment for acute angina. Generally, the sublingual administration of nitroglycerin can relieve the symptoms of acute angina within 5 to 10 minutes. The physiological therapeutic principle of nitroglycerin is mainly explained that it has the ability to release nitrogen oxide compounds (NO or NO congener) with pharmaco...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 黄薇金力李一峰金玮
Owner CHINESE NAT HUMAN GENOME CENT AT SHANGHAI
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