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Microfluidic chip, manual centrifuging device and nucleic acid detection method

A technology of microfluidic chips and centrifugal devices, applied in enzymology/microbiology devices, chemical instruments and methods, biochemical cleaning devices, etc., which can solve the limitations of the use environment, high requirements for the use environment, and difficulty in ensuring test accuracy, etc. problems, to achieve the effect of easy control, convenient use and high test accuracy

Inactive Publication Date: 2018-05-18
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the above technical solution, on the one hand, the detection device uses an automatic mechanism for testing and is supported by a support frame. During the testing process, the requirements for the use environment are relatively high, and the testing accuracy is difficult to guarantee
[0006] On the other hand, the above-mentioned detection structure adopts a nucleic acid detection optical path structure, and its inlet is connected to the outlet of the sample reaction automatic mechanism. In actual testing, because the integrated chip structure is not used, it is difficult to mix and test the reaction solution as expected.
[0007] Finally, the above-mentioned detection device is electrically connected by an automatic sample reaction mechanism and a nucleic acid detection optical path structure, which requires real-time power supply, and the use environment is limited

Method used

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  • Microfluidic chip, manual centrifuging device and nucleic acid detection method
  • Microfluidic chip, manual centrifuging device and nucleic acid detection method
  • Microfluidic chip, manual centrifuging device and nucleic acid detection method

Examples

Experimental program
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Effect test

Embodiment 1

[0067] Water-based pigments were used to simulate nucleic acid detection reaction reagents to test the ability of the manual centrifuge device and the microfluidic chip of the embodiment of the present invention to manipulate liquids.

[0068] Step S1, liquid filling process, filling the first, second and third chambers with zeolite and preloaded simulation reagent, and introducing the simulated sample mixture into the first chamber;

[0069] Step S2, the centrifugation process, fix the microfluidic chip in the manual centrifugation device, and manually pull the pull rod of the centrifugation device to make the turntable drive the chip to rotate. During the rotation, the first chamber 121, the second chamber 122, the third chamber The liquid returned to 123 will mix and eventually pool in the fourth chamber.

[0070] see Figure 10 , by observation, before centrifugation, the preloaded mock reagent and the introduced mock sample exist in their respective chambers, they are well...

Embodiment 2

[0072] Lysis buffer (0.2N NaOH, 1% SDS) was added to samples containing S. aureus. Using a microfluidic chip preloaded with zeolite and reaction reagents, the sample mixture containing lysate is then introduced into the first chamber. Repeat steps S1-S4 above, using ultrapure water as a negative control. Fix the microfluidic chip into the manual centrifuge device, and manually pull the pull rod of the centrifuge device to make the turntable drive the chip to rotate. Afterwards, fill the empty space of each chamber with mineral oil and seal each injection hole with adhesive tape. Put the portable warm patch into the lower cavity of the manual centrifugal device and the upper space of the chip, and cover the top cover of the device to start the nucleic acid amplification reaction. After 4 hours, turn on the device, and observe the fluorescence of the reaction solution in the chip chamber 4 .

[0073] see Figure 11 As shown, it is a fluorescence shot of the chip when using t...

Embodiment 3

[0075] Lysis solution (0.2N NaOH, 1% SDS) was added to samples containing Pseudomonas aeruginosa. Using a microfluidic chip preloaded with zeolite and reaction reagents, the sample mixture containing lysate is then introduced into the first chamber. Repeat steps S1-S4 above, using ultrapure water as a negative control. Use ultrapure water as a negative control. Fix the microfluidic chip into the manual centrifuge device, and manually pull the pull rod of the centrifuge device to make the turntable drive the chip to rotate. Afterwards, fill the empty space of each chamber with mineral oil and seal each injection hole with adhesive tape. Put the portable warm patch into the lower cavity of the manual centrifugal device and the upper space of the chip, and cover the top cover of the device to start the nucleic acid amplification reaction. After 4 hours, turn on the device, use the mobile phone to take pictures of the fluorescence of the reaction solution in the chip chamber 4,...

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Abstract

The invention relates to a microfluidic chip, a manual centrifuging device and a nucleic acid detection method. The microfluidic chip comprises at least one sample adding layer and a channel layer arranged at the lower side of the sample adding layer, wherein the upper surface of the channel layer is provided with a plurality of test units which are in centered distribution and are used for loading a to-be-tested reagent and transmitting the to-be-tested reagent to a chamber; each test unit comprises at least two chambers which are interconnected through a channel; and a plurality of sample adding holes corresponding to the channel layer are arranged along the central circumference of the sample adding layer for injection of a reagent into one of the chambers of a corresponding test unit.The manual centrifuging device provided by the invention is a portable device which does not rely on an external power supply, integrates nucleic acid extraction, amplification and detection by utilizing a microfluidic chip technology, realizes rapid and convenient nucleic acid analysis, and is used for rapid response and field diagnosis of emergencies like large-scale infectious diseases, food safely affairs and environmental pollution accidents.

Description

technical field [0001] The invention relates to the technical field of nucleic acid detection, in particular to a microfluidic chip, a manual centrifugal device and a nucleic acid detection method. Background technique [0002] In the prior art, commonly used nucleic acid detection methods include amplification-based rapid nucleic acid detection and hybridization-based gene chip technology, etc.; among them, the amplification-based rapid nucleic acid detection technology is favored by its advantages of sensitivity, high efficiency, rapidity, and convenience. Widely used in pathogen detection, genotyping and other fields. [0003] In the prior art, commonly used nucleic acid detection devices mainly include large-scale instruments such as PCR instruments, constant temperature nucleic acid amplification detectors, gel imaging systems, gene chip detection platforms, and genomics sequencers. These instruments have high requirements on the working environment and operators, whic...

Claims

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Application Information

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IPC IPC(8): B01L3/00C12M1/00
CPCB01L3/5027B01L3/502753B01L2200/10B01L2300/0861B01L2300/0887C12N15/1003C12Q1/6844C12Q2563/159
Inventor 孙佳姝田飞张璐刘超
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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