Method and kit for rapidly extracting nucleic acids from trace clinical samples

A sample-in-kit technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA preparation, etc., can solve the problems of inability to completely remove PCR inhibitors, high DNA purity, complicated steps, etc., and achieve operation time. Short, easy to operate, low cost effect

Inactive Publication Date: 2010-03-31
USTAR BIOTECHNOLOGIES (HANGZHOU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

spinning centrifuge column method Silica carrier adsorption most specimens DNA extraction and purification DNA purity is high, but it is time-consuming and the steps are cumbersome. Efficiency of elution is key
glass powder method Glass powder adsorption, centrifugal separation soil sample The method is simple and can be used for bacterial spore DNA in soil Extraction of PCR inhibitors cannot be completely removed

Method used

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  • Method and kit for rapidly extracting nucleic acids from trace clinical samples
  • Method and kit for rapidly extracting nucleic acids from trace clinical samples
  • Method and kit for rapidly extracting nucleic acids from trace clinical samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Sputum processing and TB DNA extraction

[0067] 1. Sputum liquefaction

[0068] Take the sputum of the tubercle bacillus (TB) subject to be tested, transfer it to a centrifuge tube, add 1 times the volume of sputum liquefaction solution, shake and mix well, heat at 95°C-100°C for 10 minutes, and cool to room temperature;

[0069] 2. Lysis

[0070] Take 1ml of liquefied sputum, add the same amount of lysate; mix well and let it stand at room temperature for 5 minutes; use the micro-clinical sample nucleic acid rapid extraction device to suck up the above mixed solution, and push out the filtrate in the suction device through a pointed pipette and discard Lose;

[0071] 3. Cleaning

[0072] Use the micro-clinical sample nucleic acid rapid extraction device to suck in the centrifuge tube containing the cleaning solution, and then push out the filtrate in the suction device through a pointed suction pipe and discard it;

[0073] 4. DNA Elution

[0074] Continue to use...

Embodiment 2

[0086] Treatment of sexually transmitted disease samples (cotton swabs) and extraction of pathogen DNA

[0087] 1. Elution

[0088] Take a cotton swab sample from a Chlamydia trachomatis (CT) patient to be tested, add 1ml of normal saline to the sample collection tube, shake and mix well to ensure that the secretions on the cotton swab are fully eluted;

[0089] 2. Lysis

[0090] Add an equal amount of lysate to the above eluent; mix well and let stand at room temperature for 5 minutes; use a micro-clinical sample nucleic acid rapid extraction device to suck up the above mixed solution, and push out the filtrate in the suction device through a pointed pipette and discard;

[0091] 3. Cleaning

[0092] Use the micro-clinical sample nucleic acid rapid extraction device to suck in the centrifuge tube containing the cleaning solution, and then push out the filtrate in the suction device through a pointed suction pipe and discard it;

[0093] 4. DNA Elution

[0094] Continue to...

Embodiment 3

[0108]Serum sample processing and extraction of hepatitis C virus (HCV) RNA

[0109] 1. Lysis

[0110] Take 200ul of HCV serum sample (thaw the frozen serum at room temperature before use, shake and mix for 10 seconds), add an equal amount of lysate, mix well, and let stand at room temperature for 5 minutes; use the micro-clinical sample nucleic acid rapid extraction device to absorb the above mixed Liquid, the filtrate in the suction device is pushed out through the pointed suction pipe and discarded;

[0111] 2. Cleaning

[0112] Use the micro-clinical sample nucleic acid rapid extraction device to suck in the centrifuge tube containing the cleaning solution, and then push out the filtrate in the suction device through a pointed suction pipe and discard it;

[0113] 3. RNA Elution

[0114] Continue to use the micro-clinical sample nucleic acid rapid extraction device to suck 100ul of the eluent, and push the liquid in the suction device into a new centrifuge tube through ...

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Abstract

The invention relates to a method for rapidly processing and extracting nucleic acids from various trace clinical samples. The method comprises the following steps: cracking cells; absorbing by a filter membrane; and leaching and extracting nucleic acids (DNA and RNA). The invention also relates to a kit for rapidly extracting nucleic acids from clinical samples and application thereof.

Description

technical field [0001] The invention relates to a rapid extraction method for nucleic acid in various clinical samples. More specifically, the present invention relates to a method for rapidly processing and extracting nucleic acids from various micro clinical samples using a device for rapidly extracting nucleic acid from a small amount of clinical samples, including the steps of cell lysis, filter membrane adsorption and elution to extract nucleic acids (DNA and RNA). The invention also relates to a kit for rapid extraction of nucleic acid from clinical samples and its application. Background technique [0002] (1) Pretreatment of clinical samples [0003] Clinically, it is often necessary to extract genomic DNA or RNA from various biological tissues or cells, and use nucleic acid amplification technology to determine its components. This is a highly sensitive and most direct detection method for auxiliary analysis and diagnosis of diseases. . Since clinical samples con...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/68C12Q1/04
Inventor 胡林尤其敏王宏莹罗英钟华燕石坚
Owner USTAR BIOTECHNOLOGIES (HANGZHOU) CO LTD
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