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

A micro-nucleic acid, filtering technology, which is used in the field of rapid processing of various nucleic acid biological samples, can solve the problems of inability to completely remove PCR inhibitors, high DNA purity, complicated steps, etc., and achieves short operation cycle, simple operation and low cost. Effect

Active Publication Date: 2012-09-19
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 TB DNA extraction

[0071] 1. Sputum liquefaction

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

[0073] 2. Suction filtration

[0074] Aspirate 1.5ml of the above-mentioned liquefied sputum using a filter-type sample processing device, disconnect the device from the second connection part, and discard the filtrate in the syringe;

[0075] 3. Washing

[0076] Reconnect the device, suction 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. Lysis

[0078] Reconnect the device, pump repeatedly in the centrifuge tube containing the lysate, 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;

...

Embodiment 2

[0092] Treatment of sexually transmitted disease samples (cotton swabs) and extraction of pathogen DNA

[0093] 1. Elution

[0094] Take the sample of the patient to be tested (CT / UU), add 1ml of normal saline to the sample collection tube, vortex 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 middle;

[0095] 2. Suction filtration

[0096] Use a filter-type sample processing device to extract all the eluent in the centrifuge tube, and finally disconnect the device from the second connection part, and discard the filtrate in the syringe;

[0097] 3. Lysis

[0098] 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 centrifuge tube at 99°C for 10 minutes, and cool to room temperature;

[0099] 4. DNA extraction

[0100] Continue to use the device to suck out the liquid...

Embodiment 3

[0116] Serum sample processing and HBV DNA extraction

[0117] 1. Lysis

[0118] Take 100ul of HBV serum sample (thaw frozen serum at room temperature before use, shake and mix for 10 seconds), add an equal amount of DNA extraction solution, mix well, put in a boiling water bath for 10 minutes, and then cool to room temperature;

[0119] 2. DNA extraction

[0120] Insert the pointed suction tube of the filter-type sample processing device into the above-mentioned boiled centrifuge tube jelly, suck out the liquid as much as possible, disconnect the device from the second connection part, and transfer the liquid in the syringe to a new one. in a centrifuge tube, set aside.

[0121] 3. Nucleic Acid Amplification

[0122] Use the HBV constant amplification reagent to amplify the extracted HBV DNA, and the specific reaction is as follows:

[0123] HBV amplification reaction solution 10 microliters

[0124] Extract DNA solution 2 microliters

[0125] wxya 2 O 8 µl

[0126] 2...

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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 amounts of nucleic acid samples in clinical biological samples by means of filtration, including enriching cells in clinical biological samples and lysing cells to extract nucleic acids. [0002] The invention also relates to a kit for rapid processing of trace 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 samples contain proteins, lipids and other substances, which will interfere with the downstream amplification reactio...

Claims

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Application Information

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Patent Type & Authority Patents(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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