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Ultra-fast real-time nucleic acid detection method for parainfluenza virus

A detection method, an ultra-fast technology, applied in the rapid detection of parainfluenza virus infection, the field of ultra-fast nucleic acid real-time fluorescence quantitative detection of parainfluenza virus, can solve the detection limit of sudden infectious diseases, the results are not easy to repeat, and amplify Problems such as complex process, to achieve the effect of valuable treatment time, short reaction time, and improved detection efficiency

Inactive Publication Date: 2018-06-15
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 parainfluenza virus 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, ...

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Abstract

The invention relates to an ultra-fast real-time nucleic acid detection method for parainfluenza virus. According to the ultra-fast real-time nucleic acid detection method, nucleic acid is extract, amplification is performed with fluorescent labeled primers in a special buffer solution in a disk-type reaction container under the catalysis of an enzyme, and the nucleic acid template content in a sample is real-timely and quantitatively detected, wherein the reaction time is substantially shortened compared to the previous fluorescence quantitative nucleic acid detection method through the special buffer solution and the disk-type reaction container, and is shortened to 5-10 min from the original time of more than 1 h, such that the rapid detection of parainfluenza virus infection is truly achieved. According to the present invention, the ultra-fast real-time nucleic acid detection method has advantages of short reaction time, simple operation, high specificity and high promotion value,and is suitable for the detection of sudden acute parainfluenza virus infection.

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 for parainfluenza virus. It can be used for the quantitative detection of nucleic acid of parainfluenza virus, especially suitable for the rapid detection of parainfluenza virus infection. 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 o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851
CPCC12Q1/6851C12Q2527/125C12Q2561/113
Inventor 冯长访
Owner 美科生物医学技术无锡有限公司
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