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A device for rapid extraction of nucleic acid and a method for rapid extraction of nucleic acid

A nucleic acid extraction device and nucleic acid technology, applied in biochemical cleaning devices, biochemical equipment and methods, enzymology/microbiology devices, etc., can solve the problems of inaccurate detection and single combination mode, and achieve safe operation and ingenious conception , the effect of simple operation

Active Publication Date: 2017-01-18
INST OF PLA FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0023] CN101684463A proposes a device for quickly extracting nucleic acid from micro-clinical samples, which consists of three parts: a pointed suction pipe, a filter with membrane and a suction device (syringe or plastic bubble), which completes the nucleic acid adsorption, cleaning and eluting process through three suctions , can quickly and easily extract nucleic acid from the sample, but the fly in the ointment is that the combination of the three parts of the pointed suction pipe, the filter with membrane and the suction device is single and fixed installation, when the suction device is made of plastic bubbles (CN101684463A figure 2 , image 3 ) the third suction for nucleic acid elution cannot actually be completed (the waste liquid from the first two suctions cannot be discharged, and the eluent with nucleic acid is mixed in the waste liquid), and when using a syringe as a suction device, the eluent with nucleic acid Dehydration into the syringe will not only contaminate the syringe to form biological waste, but also cause the nucleic acid in the trace sample to be contaminated and lost to the syringe, resulting in inaccurate detection

Method used

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  • A device for rapid extraction of nucleic acid and a method for rapid extraction of nucleic acid
  • A device for rapid extraction of nucleic acid and a method for rapid extraction of nucleic acid
  • A device for rapid extraction of nucleic acid and a method for rapid extraction of nucleic acid

Examples

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example 1

[0081] Example 1: the extraction of Salmonella DNA

[0082] This example takes the extraction of Salmonella DNA as an example to verify whether the nucleic acid extraction device and method of the present invention are effective. In this example, a total of 4 samples were extracted (1 is negative control, 2‐4 is Salmonella), and the following operations were performed respectively:

[0083] 1. Lysis

[0084] Take 500 microliters of Salmonella overnight culture bacteria solution, add the same amount of lysate (composition: NaOH, 25mmol; Tris-Hcl, 10mmol; Triton X-100, 1%; NP-40, 1%; EDTA, 0.1mMol ; Chelex‐100, 2%.); After mixing, let it stand at room temperature for 5 minutes, use the suction form of the nucleic acid extraction device, pump the mixed solution to the adsorption membrane tube, let the mixed solution pass through the adsorption membrane, and the nucleic acid-free waste fluid into the syringe cavity.

[0085] 2. Cleaning

[0086] Use the suction method of the n...

example 2

[0093] Example 2: Ring-mediated constant temperature amplification method detects Trichinella spiralis in blood

[0094] In this example, the detection of Trichinella spiralis in blood by the loop-mediated constant temperature amplification method is taken as an example to illustrate the application of the nucleic acid extraction device and method of the present invention. A total of 8 samples were tested (1-6 samples were positive infected blood samples, and 7-8 samples were negative controls), and the following detection operations were performed respectively:

[0095] 1. Lysis

[0096] Take 500 microliters of blood samples, add the same amount of lysate (composition: NaOH, 25mmol; Tris-Hcl, 10mmol; Triton X-100, 1%; NP-40, 1%; EDTA, 0.1mMol; Chelex-100 , 2%.); After mixing, let stand at room temperature for 5 minutes, use the suction form of the nucleic acid extraction device, suck the mixed solution to the adsorption membrane tube, let the mixed solution pass through the ...

example 3

[0105] Example 3: Extraction and detection of influenza A virus H1N1RNA in throat swab

[0106] This example takes the extraction and detection of influenza A virus H1N1 RNA in throat swabs as an example to illustrate that the nucleic acid extraction device and method of the present invention are also suitable for extracting RNA. There are a total of 8 test samples (1 is negative control, 2-8 are positive samples), and the following extraction and detection operations are performed respectively:

[0107] 1. Lysis

[0108] Take a throat swab sample of 200u1, add an appropriate amount of lysate (composition: NaOH, 25mmol; Tris-Hcl, 10mmol; Triton X-100, 1%; NP-40, 1%; EDTA, 0.1mMol; Chelex-100, 2% ), RNA adsorption solution (composition: add an equivalent amount of 99% ethanol to the lysate) and RNase inhibitor (composition: Carrier RNA), mix well, and let stand at room temperature for 5 minutes; use the suction form of the nucleic acid extraction device, The mixed solution is...

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Abstract

The invention relates to a device and method used for rapid extraction of nucleic acid which are mainly used for rapidly extracting nucleic acid (including RNA and DNA) from each clinical sample. The device mainly comprises an injector, a hollow adaptor, a nucleic acid adsorption membrane tube and a sampling probe, after the steps of splitting a clinical sample, sucking by virtue of an injector, adsorbing nucleic acid by virtue of a nucleic acid adsorption film, rinsing the nucleic acid adsorption film with rinsing liquid and eluting nucleic acid with eluant are carried out, a pure nucleic acid solution can be obtained to be used in a downstream experiment.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a device and method for rapidly extracting nucleic acids from various clinical samples. Background technique [0002] (1) Nucleic acid extraction [0003] Nucleic acid extraction has become the most important and basic operation in molecular biology experimental techniques, but there are currently many nucleic acid extraction schemes, and they are complicated and simple. The completion of molecular diagnosis, nucleic acid sequencing, functional genomics and other related experiments all depend on the successful extraction of nucleic acid. The traditional operation process can be fully automated, but these expensive instruments cannot be widely used. In developing countries, the vast primary medical units cannot be equipped with complex instruments, so it is necessary to provide an economical nucleic acid extraction device. [0004] Nucleic acid includes two kinds of molecules, DNA ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/00C12N15/10
CPCC12N15/1017
Inventor 袁静刘威黄留玉
Owner INST OF PLA FOR DISEASE CONTROL & PREVENTION
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