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Ultra-fast real-time nucleic acid detection method for Streptococcus pneumoniae

A Streptococcus pneumoniae, detection method technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganism determination/inspection, etc., can solve the detection limitation of sudden infectious diseases, the results are not easy to repeat, the amplification process Complex problems, to achieve the effect of valuable treatment time, short response 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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  • Ultra-fast real-time nucleic acid detection method for Streptococcus pneumoniae
  • Ultra-fast real-time nucleic acid detection method for Streptococcus pneumoniae

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Embodiment 1

[0015] Ultra-rapid quantitative detection of Streptococcus pneumoniae in human blood

[0016] 1. Pathogen DNA extraction

[0017] (1) Sample treatment: Take 100μL-500μL blood sample, add 1.4mL AS buffer, and mix repeatedly;

[0018] (2) Incubate the resuspension at 70°C for 5 minutes;

[0019] (3) Vortex for 15 seconds and place at room temperature for 1 minute. Centrifuge at the highest speed for 1 minute to precipitate;

[0020] (4) Transfer 900 μl supernatant to a 1.5ml centrifuge tube, add 100 μl impurity scavenger, vortex immediately for 1 minute, and place at room temperature for 1 minute. Centrifuge at the highest speed for 3 minutes to remove impurities;

[0021] (5) Transfer all supernatants to a 1.5ml centrifuge tube and centrifuge at top speed for 3 minutes;

[0022] (6) Transfer 210 μl of supernatant to a 1.5ml centrifuge tube, add 20 μl of proteinase K (20 mg / ml) solution, mix well, add 200 μl of binding solution, vortex for 15 seconds, and mix well. Incubat...

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Abstract

The invention relates to an ultra-fast real-time nucleic acid detection method for Streptococcus pneumoniae. 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 ina 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 thespecial 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 Streptococcus pneumoniae infectionis 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 Streptococcus pneumoniae 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 Streptococcus pneumoniae. It can be used for the quantitative detection of nucleic acid of Streptococcus pneumoniae, and is especially suitable for the rapid detection of Streptococcus pneumoniae 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 tec...

Claims

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Application Information

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IPC IPC(8): C12Q1/6851C12Q1/689C12Q1/14C12R1/46
CPCC12Q1/6851C12Q1/689C12Q2527/125C12Q2561/113C12Q2563/107
Inventor 冯长访
Owner 美科生物医学技术无锡有限公司
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