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Instrument-free nucleic acid extraction device and trace nucleic acid extraction method

A technology of nucleic acid extraction device and extraction method, which is applied in the field of non-instrument nucleic acid extraction device and trace nucleic acid extraction, and can solve problems such as inconvenient use, complicated structure, and complex structure of the device

Inactive Publication Date: 2011-05-04
USTAR BIOTECHNOLOGIES (HANGZHOU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the prior art, nucleic acid extraction devices are generally bulky and expensive
The invention application with the patent application number 200580016159.1 discloses a method and device for extracting nucleic acid. The device has a complex structure and is a high-throughput, automated instrument suitable for high-level hospitals and research institutes.
The patent application number is 200710103868.4, which discloses a nucleic acid extraction device, which contains two interconnected containers, a mechanism for sending in / out solution, and two pressurization and decompression devices. The nucleic acid extraction device of this invention is relatively It is small, but the structure is still relatively complicated, and it is not very convenient to use: it is necessary to use the mechanism of sending in / out solution to send the sample to be tested, and it is necessary to use the pressurization and decompression device to simultaneously pressurize and decompress at the same time. pressure operation

Method used

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  • Instrument-free nucleic acid extraction device and trace nucleic acid extraction method
  • Instrument-free nucleic acid extraction device and trace nucleic acid extraction method
  • Instrument-free nucleic acid extraction device and trace nucleic acid extraction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Example 1 Treatment of sputum and extraction of pulmonary tuberculosis DNA

[0085] 1. Liquefaction of sputum

[0086] Take the sputum of the tubercle bacillus (TB) to be examined, transfer it to a centrifuge tube, add 1 volume of sodium hydroxide sputum liquefaction solution, shake and mix thoroughly, heat at 95℃-100℃ for 10 minutes, and cool to room temperature ;

[0087] 2. Cracking

[0088] Take 1ml of liquefied sputum and add the same amount of lysate; after mixing, let it stand at room temperature for 5 minutes; use the micro-clinical sample nucleic acid rapid extraction device to suck the above-mentioned mixed solution, and push out the filtrate in the suction device and discard;

[0089] 3. Cleaning

[0090] Use the rapid extraction device of micro clinical sample nucleic acid to suck in the centrifuge tube containing the cleaning solution, push out the filtrate in the suction device and discard;

[0091] 4. DNA elution

[0092] Continue to use the micro clinical sample nuc...

Embodiment 2

[0104] Example 2 Treatment of sexually transmitted disease samples (cotton swabs) and extraction of pathogen DNA

[0105] 1. Elution

[0106] Take a cotton swab sample from a patient to be tested for chlamydia (CT), add 1ml of normal saline into the sample collection tube, shake and mix well to ensure that the secretions on the cotton swab are fully eluted;

[0107] 2. Cracking

[0108] Add an equal amount of lysate to the above eluent; let it stand for 5 minutes at room temperature after mixing; use a micro-clinical sample nucleic acid rapid extraction device to absorb the above mixed solution, and push out the filtrate in the suction device to discard;

[0109] 3. Cleaning

[0110] Use the rapid extraction device of micro clinical sample nucleic acid to suck in the centrifuge tube containing the cleaning solution, push out the filtrate in the suction device and discard;

[0111] 4. DNA elution

[0112] Continue to use the micro-clinical sample nucleic acid rapid extraction device to suck...

Embodiment 3

[0130] Serum sample processing and hepatitis C virus (HCV) RNA extraction

[0131] 1. Cracking

[0132] Take 200ul of HCV serum sample (thawed at room temperature before using the frozen serum, shake and mix for 10 seconds), add the same amount of lysate, mix well, and let stand at room temperature for 5 minutes; use a micro clinical sample nucleic acid rapid extraction device to absorb the mixture Liquid, push out the filtrate in the suction device and discard;

[0133] 2. Cleaning

[0134] Use the rapid extraction device of micro clinical sample nucleic acid to suck in the centrifuge tube containing the cleaning solution, push out the filtrate in the suction device and discard;

[0135] 3. RNA elution

[0136] Continue to use the micro-clinical sample nucleic acid rapid extraction device to suck 100ul of eluent, and push the liquid in the suction device to a new centrifuge tube for use;

[0137] 4. Nucleic acid amplification

[0138] Use RT-HCV constant temperature amplification reagent...

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Abstract

The invention relates to an instrument-free nucleic acid extraction device and a trace nucleic acid extraction method. The trace nucleic acid extraction device comprises a tubular main body 1, wherein one end of the tubular main body is provided with a joint 2 connected with a suction device and the other end of the tubular main body is provided with a plug 3; and a filter element device 4 for combining nucleic acid is arranged inside the tubular main body or the plug. The trace nucleic acid extraction device replaces a high-rotating speed centrifugal machine, can achieve good separation effect in a heterogeneous liquid phase system, is low in cost, easy to operate and short in operating time, and can be widely applied to rapid nucleic acid diagnosis in primary hospitals and emergency rooms under relatively bad experiment conditions. A nucleic acid detection method is a simple means of extracting a biological sample of nucleic acid, the purity of the extracted sample meets the requirement on nucleic acid amplification, other instruments are not needed, the device is used at one time, and the method can be widely applied to extracting pathogen or host nucleic acid, and is used for clinical detection and diagnosis.

Description

Technical field [0001] The invention relates to an instrumentless micro nucleic acid extraction device and a micro nucleic acid extraction method. Background technique [0002] It is often necessary to extract genomic DNA or RNA from various biological tissues or cells in clinical practice, 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 the clinical specimens contain proteins, lipids and other substances, which will interfere with the downstream amplification reaction, biological sample processing and nucleic acid extraction must be carried out before the detection reaction. Since the processing and storage of samples have a greater impact on DNA and RNA (especially RNA), proper pretreatment and storage of nucleic acid specimens have a decisive effect on the successful extraction of nucleic acid templates. Common clinical specimens in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/68C12Q1/70C12R1/93C12R1/01C12R1/32
Inventor 胡林尤其敏徐高连石坚王宏莹钟华燕贾子冬祁军
Owner USTAR BIOTECHNOLOGIES (HANGZHOU) CO LTD
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