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Method for quickly processing filter-type micro nucleic acid clinical samples

A technology of trace nucleic acid and processing method, which is applied in the field of rapid processing of various nucleic acid biological samples, which can solve the problems of inability to completely remove PCR inhibitors, high DNA purity, cumbersome steps, etc., and achieve short operation cycle, simple operation, and low cost Effect

Active Publication Date: 2009-09-23
USTAR BIOTECHNOLOGIES (HANGZHOU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

spin column Law 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 suction attached, centrifuged Leave soil sample The method is simple and can be used for bacterial spore DNA in soil extraction, cannot completely remove PCR inhibitors

Method used

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  • Method for quickly processing filter-type micro nucleic acid clinical samples
  • Method for quickly processing filter-type micro nucleic acid clinical samples
  • Method for quickly processing filter-type micro nucleic acid clinical samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Sputum processing and TR DNA extraction

[0071] 1. Liquefaction of sputum

[0072] Take the sputum of the TB candidate, transfer it to a centrifuge tube, add 1 volume of sputum liquefaction solution, shake and mix thoroughly;

[0073] 2. Suction filtration

[0074] Use a filtering sample processing device to suck 1.5ml of the above-mentioned liquefied sputum, disconnect the device from the second connection part, and discard the filtrate in the syringe;

[0075] 3. Washing

[0076] Reconnect the device, aspirate in the centrifuge tube containing the cleaning solution (normal saline), and finally disconnect the device from the second connection part, and discard the filtrate in the syringe;

[0077] 4. Cracking

[0078] Reconnect the device, repeatedly pump in the centrifuge tube with lysis solution, and finally push all the liquid in the injector back into the centrifuge tube, heat the centrifuge tube at 99°C for 10 minutes, and cool to room temperature;

[0079] 5. DNA ex...

Embodiment 2

[0093] Treatment of sexually transmitted disease samples (cotton swabs) and extraction of pathogen DNA

[0094] 1. Elution

[0095] Take a sample of the patient to be tested (CT / UU), add 1ml of saline to the sample collection tube, shake and mix to ensure that the secretions on the cotton swab are fully eluted, and transfer all the eluate to a 1.5ml centrifuge tube in;

[0096] 2. Suction filtration

[0097] Use a filtering sample processing device to extract all the eluate in the centrifuge tube, and finally disconnect the device from the second connection part, and discard the filtrate in the syringe;

[0098] 3. Cracking

[0099] Reconnect the device, use the device to repeatedly suck the lysate in the centrifuge tube, and finally push all the liquid in the injector back into the centrifuge tube, heat the tube at 99°C for 10 minutes, and cool to room temperature;

[0100] 4. DNA extraction

[0101] Continue to use the device to suck out the liquid in the heated tube, disconnect...

Embodiment 3

[0118] Serum sample processing and HBV DNA extraction

[0119] 1. Cracking

[0120] Take 100ul HBV serum sample (thawed at room temperature before using the frozen serum, shake and mix for 10 seconds), add the same amount of DNA extract, mix well, boil water for 10 minutes, then cool to room temperature

[0121] 2. DNA extraction

[0122] Deepen the pointed suction tube of the filtering sample processing device into the boiled centrifuge tube gel, suck out the liquid and try to suck it up, disconnect the device from the second connection part, and transfer the liquid in the syringe to a new one. In the centrifuge tube, set aside.

[0123] 3. Nucleic acid amplification

[0124] Apply HBV constant stability amplification reagent to amplify the extracted HBV DNA, the specific reaction is as follows:

[0125] HBV amplification reaction solution 10 μl

[0126] Extract DNA solution 2 μl

[0127] ddH 2 O 8 μl

[0128]

[0129] The total volume is 20 microliters

...

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Abstract

The invention relates to a method for quickly processing various nucleic acid biological samples, in particular to a method for quickly processing and extracting micro nucleic acid samples in clinical biological samples by adopting a filter-type nucleic acid extraction device consisting of a syringe filter and a clinical disposable syringe. The method can finish all steps including enrichment, cleaning, cracking and nucleic acid extraction of host cells, bacteria and viruses in cells in the clinical biological samples. The invention also relates to a kit for quickly processing the micro nucleic acid clinical samples and application thereof.

Description

Technical field [0001] The invention relates to a rapid processing method for various nucleic acid biological samples. More specifically, the present invention relates to a method for rapidly processing and extracting trace nucleic acid samples in clinical biological samples by filtering, including enrichment of cells in clinical biological samples and lysis of cells to extract nucleic acids. [0002] The invention also relates to a kit for rapidly processing micro nucleic acid clinical samples and applications thereof. Background technique [0003] Clinically, it is often necessary to extract genomic DNA 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 specimens contain proteins, lipids and other substances, which will interfere with the downstream amplification reaction, biological sample...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/68C12Q1/02C12M1/12
Inventor 胡林尤其敏王宏莹石翊轩
Owner USTAR BIOTECHNOLOGIES (HANGZHOU) CO LTD
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