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Primer composition for detecting nitroglycerin drug related gene and kit

A technology of primer composition and nitroglycerin, which is applied in the direction of recombinant DNA technology, microbial measurement/testing, biochemical equipment and methods, etc., can solve the problems of limitation and high price, and achieve rapid detection, simple method and accurate results Effect

Inactive Publication Date: 2018-07-24
宁波美丽人生医学检验所有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, similar kits currently on the market are expensive and have limited applications

Method used

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  • Primer composition for detecting nitroglycerin drug related gene and kit
  • Primer composition for detecting nitroglycerin drug related gene and kit
  • Primer composition for detecting nitroglycerin drug related gene and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] 1. Sample processing and nucleic acid extraction:

[0050] Use a commercial DNA extraction kit to process the sample. For specific operations, refer to the kit instruction manual. The concentration and purity of the extracted DNA need to be measured with a UV spectrophotometer. The DNA OD 260 / OD 280 The value of DNA should be between 1.8 and 2.0, and the concentration should be between 5 and 50 ng / μL. If the DNA quality of the sample is unqualified, it should not be used for detection. If it is lower than 5 ng / μL, the nucleic acid should be extracted again, and if it is higher than 50 ng / μL, it should be properly diluted. To the specified concentration range, the extracted DNA should be detected immediately, otherwise it should be stored below -20°C, and the storage time should not exceed 6 months. The quality control product of the kit was thawed at room temperature before use, vortexed for 10 seconds, and centrifuged at 2000rpm for 15 seconds before use.

[0051] 2...

Embodiment 2

[0069] The primer composition for detecting nitroglycerin drug-related genes includes an ALDH2 primer set, and the ALDH2 primer set includes the following primers:

[0070] P1-FW: 5'-CGGGCTGCAGGCATACACAG-3';

[0071] P1-FS: 5'-CGGGCTGCAGGCATACACAA-3';

[0072] P1-R: 5'-GTCCTGAACTTCCAGCAG-3'.

[0073] Application of the above primer composition in the preparation of reagents for detecting gene polymorphisms of nitroglycerin drug-related genes.

[0074] The kit for detecting nitroglycerin drug-related genes includes the above-mentioned primer composition and PCR reaction liquid, and the PCR reaction liquid includes PCR buffer, Taq enzyme, MgCl 2 , dNTPs and SYBR Green I dye.

Embodiment 3

[0076] The primer composition for detecting nitroglycerin drug-related genes includes an ALDH2 primer set, and the ALDH2 primer set includes the following primers:

[0077] P1-FW: 5'-CGGGCTGCAGGCATACACAG-3';

[0078] P1-FS: 5'-CGGGCTGCAGGCATACACAA-3';

[0079] P1-R: 5'-GTCCTGAACTTCCAGCAG-3'.

[0080] The primer composition also includes an internal control primer set, which includes the following primers:

[0081] CF: 5'-AGCAAGCAGGAGTATGACG-3';

[0082] CR: 5'-GAAAGGGTGTAACGCAACT-3'.

[0083] Application of the above primer composition in the preparation of reagents for detecting gene polymorphisms of nitroglycerin drug-related genes.

[0084] The kit for detecting nitroglycerin drug-related genes includes the above-mentioned primer composition and PCR reaction liquid, and the PCR reaction liquid includes PCR buffer, Taq enzyme, MgCl 2 , dNTPs and SYBR Green I dye.

[0085] The kit also includes a positive control solution and a negative control solution.

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Abstract

The invention relates to a primer composition for detecting a nitroglycerin drug related gene and a kit. The primer composition comprises a primer group ALDH2 (acetaldehyde dehydrogenase 2). The kit comprises the primer composition. Compared with the prior art, the primer composition and the kit have the advantages that the kit capable of rapidly, sensitively, simply and conveniently detecting gene polymorphism of a nitroglycerin drug treatment effect related gene (ALDH2) is obtained by adopting an ARMS (amplification refractory mutation system) technology and SYBR dye combined method; the kitcomprises a specific ARMS detection primer, an internal control primer and a PCR (polymerase chain reaction) solution, the detecting cost is reduced greatly through designing the ARMS primer and changing an Scorpions probe into an SYBR dye, thus, the kit is more applicable to the detection on polymorphism of the gene ALDH2 of a Chinese patient, has the advantages of high detection speed, high sensitivity, high specificity, simple method and accurate result and is applicable to popularization and application.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and relates to a primer composition and a kit for detecting nitroglycerin drug-related genes. Background technique [0002] Nitroglycerin (Glycery1 Trinitrate, GTN) is the drug of choice for the treatment of angina pectoris. It can directly relax smooth muscle, especially vascular smooth muscle, and inhibit the spasm and expansion of local stenosis of coronary arteries. It is the first nitrate ester used to treat coronary heart disease. drug. [0003] Aldehyde dehydrogenase 2 (ALDH2) is a key enzyme in the process of converting nitroglycerin into nitric oxide. Its coding gene is located at 12q24 on the chromosome, and there is a high degree of genetic polymorphism in this gene. ALDH2 can catalyze the hydrolysis of nitroglycerin to generate 1,2-dinitroglycerin and nitrite, and then generate NO to play a vasodilation effect. The study found that the ALDH2 gene has a G-A point mutation at p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6883C12Q2600/106C12Q2600/156C12Q2535/137C12Q2563/107C12Q2545/114
Inventor 田晓丽
Owner 宁波美丽人生医学检验所有限公司
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