Reaction tube used in loop-mediated isothermal amplification technique and use method thereof

A loop-mediated isothermal amplification reaction technology, applied in the field of temperature amplification, can solve problems such as aerosol pollution in LAMP reaction, and achieve the effects of low cost, reduced pollution, and guaranteed detection.

Inactive Publication Date: 2010-05-26
GUANGZHOU HUAFENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to address the deficiencies of the prior art, to provide a cost increase of less than 3%, which can solve the problem of

Method used

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  • Reaction tube used in loop-mediated isothermal amplification technique and use method thereof
  • Reaction tube used in loop-mediated isothermal amplification technique and use method thereof
  • Reaction tube used in loop-mediated isothermal amplification technique and use method thereof

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Experimental program
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Effect test

Embodiment 1

[0038] Such as image 3 As shown, the reaction tube of this embodiment is composed of a tube body and a tube cover. The lower part of the inner cavity of the tube is provided with a longitudinally extending partition separating it into two cavities A and B, wherein: cavity A There are 22.0 microliters of LAMP reaction solution and 0.5 microliters of Bst DNA polymerase inside, and the upper layer of the liquid is sealed with paraffin; the cavity B is filled with 2.0 microliters of LAMP reaction chromogenic solution, and the upper layer of the liquid is also sealed with paraffin, and the reaction Store the tube at -20°C.

Embodiment 2

[0039] The detection of embodiment 2 Mycobacterium tuberculosis

[0040] This embodiment adopts the reaction tube prepared in Example 1 (primers in the reaction solution are selected from a known primer group for detecting Mycobacterium tuberculosis), and detects Mycobacterium tuberculosis by LAMP, and a positive control group and a negative control group are provided at the same time. Process such as Figure 4 As shown, it specifically includes the following steps:

[0041] Take three reaction tubes prepared in Example 1, and add 2.5 μL each of negative control sample, positive control sample and sample to be tested respectively. When adding the sample, the tip of the pipette penetrates the protective liquid layer, and the sample is added to the large cavity of the Huafeng tube. Tightly cap and label the tube and move to the reaction area.

[0042] The above reaction tubes were reacted at a constant temperature of 65° C. for 1 h.

[0043] At the end of the reaction, shake ...

Embodiment 3

[0046] The detection of embodiment 3 Salmonella

[0047]This embodiment adopts the reaction tube prepared in Example 1 (primers in the reaction solution are selected from a known primer group for Salmonella detection) to detect Salmonella by LAMP, and a positive control group and a negative control group are provided at the same time. The process is as follows: Figure 4 As shown, it specifically includes the following steps:

[0048] Take three reaction tubes prepared in Example 1, and add 2.5 μL each of negative control sample, positive control sample and sample to be tested respectively. When adding the sample, the tip of the pipette penetrates the protective liquid layer, and the sample is added to the large cavity of the Huafeng tube. Tightly cap and label the tube and move to the reaction area.

[0049] The above reaction tubes were reacted at a constant temperature of 65° C. for 1 h.

[0050] At the end of the reaction, shake the reaction tube upside down once, and th...

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Abstract

The invention discloses a reaction tube used in a loop-mediated isothermal amplification technique, wherein the reaction tube is composed of two parts of a tube body and a tube cover; the lower part of the inner cavity of the tube body is provided with a longitudinally outspread clapboard which separates the inner cavity of the tube body into two cavities of A and B; the cavity A and the cavity B are respectively provided with loop-mediated isothermal amplification reaction working solution or developing solution, the upper layers of the liquid in the two cavities are both sealed up by paraffin wax. Through the design of the clapboard and the paraffin wax, the reaction tube in the invention effectively separates the developing solution and the working solution, thus not only ensuring to relatively maintain complete closure in the storage and transportation process, but also having no need to open the tube cover to realize the developing reaction and greatly reduce the pollution of aerosol. The reaction tube of the invention ensures rapid and efficient detection of loop-mediated isothermal amplification, and has low cost and is beneficial for large-scale promotion.

Description

technical field [0001] The invention relates to a loop-mediated isothermal amplification technique, in particular to a reaction tube which can be used in the loop-mediated isothermal amplification technique and a method for using the same. Background technique [0002] The emergence of PCR technology has brought the medical detection of nucleic acid level into a new stage. However, as PCR technology is more and more widely used, it is also facing severe challenges. Due to the high sensitivity of nucleic acid amplification technology, PCR has emerged. The serious problem of aerosol pollution has led to an increase in the false positive rate of PCR test results. After years of hard work, researchers have invented fluorescent quantitative PCR, which completely solved the problem of PCR aerosol pollution. However, the instrument cost of fluorescent quantitative PCR is too high to be suitable for on-site rapid detection, so this technology cannot be promoted on a large scale. ...

Claims

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Application Information

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IPC IPC(8): C12M1/24C12Q1/68
CPCC12M23/38
Inventor 曹以诚陈洵郭庆辉李晓娜谢丽丽杜正平
Owner GUANGZHOU HUAFENG BIOTECH
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