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Ultrafast detection method for nucleic acid

A detection method, ultra-fast technology, applied in fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of detection limitations of sudden infectious diseases, difficult to repeat results, complicated amplification process, etc., to achieve valuable treatment time and response The effect of shortening the time and improving the detection efficiency

Inactive Publication Date: 2012-09-19
美科生物医学技术无锡有限公司
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Problems solved by technology

However, nucleic acid detection technology still faces many challenges. In the existing nucleic acid detection technology, the entire amplification time is more than one hour, and the amplification process is relatively complicated. The results are not easy to repeat, especially in the detection of sudden infectious diseases. restricted in terms of

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Embodiment 1

[0015] Ultra-rapid quantitative detection of hsa-miR-29a miRNA in human blood

[0016] 1. Total RNA extraction

[0017] (1) Sample treatment: Take 100 μL ~ 500 μL blood sample, add 1 mL TRIzol reagent, and mix repeatedly;

[0018] (2) RNA isolation: place the above sample solution at room temperature for 10 minutes, add 200 μL of chloroform, shake vigorously for about 1 minute, let stand at room temperature for 5 minutes, centrifuge at 12,000 g at 4°C for 15 minutes, carefully take out the sample, and observe that the sample is divided into three layers. The top layer contains RNA components;

[0019] (3) RNA precipitation: Carefully pipette about 450 μL of the supernatant into a new centrifuge tube containing 600 μL of ice-cold isopropanol, mix well, and centrifuge at 12,000 g at 4°C for 10 minutes;

[0020] (4) RNA washing: remove the supernatant, add 500 μL of frozen 75% ethanol, bounce the precipitate, centrifuge at 12000g at 4°C for 5 minutes, remove the supernatant, ce...

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Abstract

The invention relates to an ultrafast detection method for nucleic acid. The method is characterized by extracting nucleic acid, performing an amplification in a special buffer solution and a disk type reactor under the catalysis of an enzyme using a fluorescent-labeled primer and detecting quantitatively the content of nucleic acid templates in samples in real time. The special buffer solution and the disk type reactor enable the reaction time to be shortened greatly compared with that of conventional fluorescent quantitative detection method for the nucleic acid, to be shortened from an original time of over one hour to a present time of 5-10 minutes, so that a fast detection for pathogen infections is really realized. The method has the advantages of short reaction time, simple process and high specificity, is particularly suitable for detecting emerging acute infectious diseases, and has high popularization value.

Description

technical field [0001] The invention relates to a nucleic acid detection method, in particular to an ultra-fast nucleic acid real-time fluorescence quantitative detection method. It can be widely used in nucleic acid quantitative detection of diseases, and is especially suitable for rapid detection of sudden infectious diseases. Background technique [0002] In 1985, Mullis and others from the Human Genetics Laboratory of PE-Cetus Company in the United States invented the epoch-making polymerase chain reaction (Polymerase Chain Reaction, PCR), which allows people to purposefully and infinitely amplify specific nucleic acid fragments in vitro. Its principle is similar to the in vivo replication of DNA, except that a suitable condition is provided for the in vitro synthesis of DNA in a centrifuge tube. After years of development, this technology has become very mature and is now the most important and commonly used technology in the field of molecular biology research and cli...

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64
Inventor 冯长访肖乐义王保君康飞蔡颖颖巩红铃谢侃冯晓程陶建国
Owner 美科生物医学技术无锡有限公司
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