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Anti-pollution portable gene detection method and device

A gene detection, portable technology, applied in the field of detection of infectious disease pathogens, can solve the problems of fluorescent probes, expensive equipment, complex device structure, etc., to achieve the effect of less labor costs, simple operation, and reduced pollution

Inactive Publication Date: 2015-12-30
GUANGZHOU HEAS BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these devices are complex in structure, require the use of fluorescent probes, and the equipment is expensive
In addition, special equipment and skilled inspection techniques are required in the detection of nucleic acid amplification products.

Method used

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  • Anti-pollution portable gene detection method and device
  • Anti-pollution portable gene detection method and device

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Isothermal Amplification Detection of HPV18 Virus

[0035] 1) The specific components of the recombinase-dependent constant temperature amplification reaction reagent are as follows:

[0036]

[0037]2) Prepare two tubes of reagents according to the reaction system in Example 1, and add HPV18 forward primer, reverse primer and fluorescent probe to the system. The two tube systems are respectively numbered as reaction tube No. 1 and reaction tube No. 2 , Then, add 2ulhela cells to No. 1 reaction tube, add 2ul human genomic DNA to No. 2 reaction tube as a negative control, and make up the system to 20ul with sterile water.

[0038] Forward primer (biotin marker): 20pmol

[0039] Reverse primer: 20pmol

[0040] FITC-labeled probe 15pmol

[0041] 3) After mixing, put the No. 1 reaction tube and No. 2 reaction tube into a 37°C constant temperature water bath for reaction, take out the reaction tube after 20 minutes, and denature at 95°C for 10 minutes. Put the two rea...

Embodiment 2

[0043] General PCR amplification detection of HPV18 virus

[0044] 1) The specific ingredients are as follows:

[0045]

[0046] 2) Prepare two tubes of reagents according to the reaction system in Example 2. The two tube systems are respectively numbered No. 1 reaction tube and No. 2 reaction tube. Then, add 2ulhela cells to No. 1 reaction tube, and add 2ul of human genomic DNA was used as a negative control, and the system was made up to 20ul with sterile water.

[0047] Forward primer (biotin marker): 4 pmol

[0048] Reverse primer: 4pmol

[0049] 3) Set the amplification conditions on the RCR amplification instrument as follows:

[0050] 95°C———15min94°C——30s; 55°C——30s; 72°C——45s72°C——10min

[0051] (40 cycles)

[0052] 4) Detection:

[0053] Put the two reaction tubes into the two airtight devices of the present invention respectively, cover the screw caps, and then squeeze the outer wall of the solution storage room, the result shows that there is a C line (qua...

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Abstract

The invention relates to an anti-pollution portable gene detection method and device. The operation procedure comprises the steps that a PCR pipe containing amplification products is put into a device to conduct sealing, and by means of a series of simple manual operations, a detection result visible by eyes is obtained; target nucleic acid amplification products can be detected rapidly, nucleic acid amplification products can be prevented from being polluted, and a false positive result is avoided. Application can be achieved in the aspects of species identification on the gene level in the fields such as pathogen detection on clinical infectious diseases, food pathogenic microbe detection, agriculture, industry, customs and animal husbandry.

Description

technical field [0001] The present invention relates to an anti-pollution portable gene detection method and device, more specifically, to a method for rapidly detecting target nucleic acid sequence amplification in an anti-pollution portable gene detection device using nucleic acid thin film chromatography (test strip) technology. The invention also relates to a kit for rapidly detecting target nucleic acid amplification products using the anti-pollution portable gene detection device and its use in detecting infectious disease pathogens. Background technique [0002] Nucleic acid research has a history of more than 100 years. In the late 1960s and early 1970s, people devoted themselves to the study of gene in vitro isolation technology. In 1983, American scientist Kary Mullis drove on the winding interstate highway and gave birth to PCR. prototype. After two years of hard work, the idea of ​​PCR was confirmed experimentally, and the first PCR patent was applied for in 198...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12M1/34C12Q1/68
CPCC12Q1/686C12Q2547/101C12Q2565/625C12Q2563/179
Inventor 陈华云刘淑园陈嘉昌肖湘文丁渭曾烨叶映玲陆嫚云黄爽彭俊然
Owner GUANGZHOU HEAS BIOTECH CO LTD
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