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Fast detection method of microRNA-208b

A detection method and rapid technology, applied in the direction of microbial determination/inspection, biochemical equipment and methods, etc., can solve the problems of shortening the detection time of microRNA-208b myocardial injury markers, long detection time, heavy workload, etc., to achieve shortened The detection time, the detection method is simple, and the effect of less time is used

Inactive Publication Date: 2016-12-07
WUXI NO 2 PEOPLES HOSPITAL
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Problems solved by technology

[0005] In view of the above-mentioned problems, the present invention provides a rapid detection method of microRNA-208b, which is applied to the detection of myocardial injury markers of acute myocardial infarction, so as to overcome the detection method of microRNA-208b in the prior art. The markers lead to a long detection time and a large workload, which greatly shortens the detection time of microRNA-208b, a myocardial injury marker, and simplifies the detection method of the above-mentioned microRNA-208b, a myocardial injury marker. On the basis of quality, the detection efficiency is improved

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Embodiment 1

[0025] The rapid detection method of microRNA-208b provided by Example 1 of the present invention comprises:

[0026] Preparation: prepare patient serum, RNase-free water, RNase inhibitor at a concentration of 18uM / uL and DSN enzyme at a concentration of 0.18uM / uL;

[0027] Design probes: Design a single-stranded DNA sequence (ACAAAC CTT TTG TTC GTC TTAT) that is completely complementary to the microRNA-microRNA-208b sequence of the target to be tested, and then modify the fluorescent groups at both ends of the single-stranded DNA sequence (5 '-TAMRA) and a fluorescent quencher (Eclipse-3'), to obtain a Taqman probe with a concentration of 200nmol / ul——Probe microRNA-208b: 5'-TAMRA-ACAAAC CTT TTG TTC GTC TTAT-Eclipse-3' ;

[0028] Reaction: Mix 50 ul of patient serum, 139 ul of RNase-free water, 1 ul of DSN enzyme, 10 ul of Taqman probe Probe microRNA-208b and 0.5 ul of RNase inhibitor to prepare a reaction volume at a reaction temperature of 85°C React for 25 minutes;

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Abstract

The invention relates to a fast detection method of microRNA-208b, and belongs to the technical field of biological detection. The fast detection method of microRNA-208b is characterized in that according to the sequence of a target microRNA-208b to be tested, a completely complementary single-chain DNA (deoxyribonucleic acid) sequence is designed; then, a fluorophore and a fluorescent quencher are respectively modified at two ends of the single-chain DNA sequence, so that a Taqman probe corresponding to the target microRNA-208b to be tested is obtained; then, patient serum, RNase-free water, DSN enzymes, Taqman probes and RNA enzyme inhibitors take reaction according to a certain proportion under the specific condition; under the stimulation effect of laser, the fluorescence intensity of the reaction liquid is detected; the detection result of the target microRNA-208b to be tested is obtained. The detection method has the advantages that the detection time of the myocardial damage landmark microRNA-208b is greatly shortened; on the basis of ensuring the detection quality, the detection efficiency is improved.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a rapid detection method for microRNA-208b. Background technique [0002] Acute myocardial infarction is myocardial necrosis caused by acute and persistent coronary ischemia and hypoxia. Clinically, there are often severe and persistent substernal pains, which cannot be completely relieved by rest and nitrates, accompanied by increased serum myocardial enzyme activity and progressive ECG changes, which may be complicated by arrhythmia, shock or heart failure, often life-threatening. The disease is most common in Europe and the United States, and about 1.5 million people in the United States suffer from myocardial infarction every year. China has shown a clear upward trend in recent years, with at least 500,000 new cases and at least 2 million existing cases each year. [0003] It can be seen that the early detection of acute myocardial infarction is particularly important. I...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/68C12Q1/6883C12Q2600/178C12Q2563/107C12Q2561/101
Inventor 韩志君蒋廷旺任天丽黄红宇高明珠魏云玉
Owner WUXI NO 2 PEOPLES HOSPITAL
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