A portable nucleic acid detection platform suitable for PCR chromatography

A detection platform and a portable technology, applied in specific-purpose bioreactors/fermenters, biochemical equipment and methods, biochemical instruments, etc., can solve problems such as pollution, high cost of fluorescent quantitative PCR instruments, and inapplicability, and achieve improved The effect of detection efficiency, intuitive readability and simple structure

Active Publication Date: 2021-03-26
宝瑞源生物技术(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For example, in terms of primary medical PCR detection, the current PCR detection method can only be used in large-scale medical institutions. , nucleic acid amplification and other intervals are strictly separated to avoid aerosol pollution, and are not suitable for detection in grassroots or remote areas;
[0005] In addition, in the application process of PCR detection technology in customs, agriculture, epidemic prevention, etc., in many cases, on-site sampling, on-site processing and interpretation of the results are required. The current technology cannot meet these requirements. It is necessary to invent and develop corresponding portable nucleic acid detection devices to solve the problem. these questions

Method used

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  • A portable nucleic acid detection platform suitable for PCR chromatography
  • A portable nucleic acid detection platform suitable for PCR chromatography
  • A portable nucleic acid detection platform suitable for PCR chromatography

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The platform introduced by the invention is used to detect the carrying rate of the epidemic Japanese encephalitis virus carried by pigs, and the field detection is carried out in the farm. The steps are as follows.

[0029] 1) Take 5ul of pig ear venous blood with a disposable syringe, add it to the sampling tube, add 30-50ul of sterilized liquid paraffin, centrifuge instantaneously, let the blood pass through the valve, and mix with the pre-installed RNA nucleic acid micro release agent.

[0030] The composition of described RNA trace nucleic acid releasing machine is: the BSA of the KCl of 500mM, 20%Triton X-100, 100mg / ml proteinase K, the NaOH of 120mM, the DMSO of pH value 6, final concentration 10%. Spin the above sampling tube into the PCR instrument and run the lysis program: 90°C for 10min-4°C for 5min.

[0031] 2) After the lysis procedure is completed, screw the sampling tube vertically into the reaction tube. The PCR amplification reagent pre-installed in t...

Embodiment 2

[0040] Detection of Shigella on surfaces of food production equipment using the platform introduced in this invention.

[0041] According to the characteristics of the present invention, its steps can be divided into the following steps.

[0042]1) Sampling: Use a disposable sterilized cotton swab, dip it in sterile water, smear the sample on the specified area, and then drop the water on the cotton swab into the sampling tube, about 5-10ul. Add 30-50ul sterile liquid paraffin for blocking.

[0043] 2) Lysis: Centrifuge briefly to make the blood pass through the valve and mix with the pre-installed RNA nucleic acid micro release agent.

[0044] The composition of described DNA trace nucleic acid releasing machine is: the BSA of the NaOH of the KCl of 50mM, 25%Triton X-100, 10mg / ml, the NaOH of 120mM, the DMSO of pH value 8, final concentration 15%. Spin the above sampling tube into the PCR instrument and run the lysis program: 90°C for 10min-4°C for 5min.

[0045] 3) After ...

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PUM

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Abstract

The invention relates to a portable nucleic acid detection platform applicable to a PCR (Polymerase Chain Reaction) chromatographic method. The portable nucleic acid detection platform is characterized in that reaction consumables comprise a sampling tube, a reaction tube and a detection card which are mounted in a sequential spiral sleeve manner; the sampling tube is provided with a sealed tube cover; a pyramid structure for puncturing through a bottom membrane of the sampling tube is arranged in the middle of the tube; an internal screw and another pyramid structure are arranged inside a detection card sample feeding hole; the reaction tube can be screwed in and punctured through, so as to ensure that a reaction environment is sealed and detection results are not polluted. The inventionfurther relates to a concept map of a portable temperature control circulator. The portable nucleic acid detection platform has the characteristics of small volume, rapid circulation and the like, after being combined with the consumables and corresponding pre-loaded reaction reagents, detection is implemented, pollution in the reaction process is avoided, results are direct and visible, more importantly, and the platform is convenient to carry over, very applicable to outdoor and on-site detection and applicable to point-of-care testing (POCT) popularization.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and relates to a portable detection platform integrated and improved by multiple technologies such as rapid processing of trace samples, rapid amplification and immunochromatographic interpretation, and is especially suitable for outdoor rapid detection. Background technique [0002] PCR, also known as polymerase chain reaction (Polymerase Chain Reaction), is an in vitro nucleic acid amplification technique developed in the mid-1980s. It has outstanding advantages such as specificity, sensitivity, high yield, rapidity, simplicity, good repeatability, and easy automation; it can amplify the target gene or a certain DNA fragment to be studied to 100,000 or even within a few hours in a test tube. A million times, so that the naked eye can directly observe and judge. The advent of PCR has caused a revolution in molecular biology, which has greatly accelerated the process of research on ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/00C12M1/38C12M1/34
Inventor 李雨峰陈立柱杨海侠
Owner 宝瑞源生物技术(北京)有限公司
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