CN212560268U

CN212560268U - Double-layer micro-fluidic chip

Info

Publication number: CN212560268U
Application number: CN202021123672.9U
Authority: CN
Inventors: 韩琳; 强乐; 张宇; 刘宏; 王敏
Original assignee: Shandong University
Current assignee: Shandong University
Priority date: 2020-06-17
Filing date: 2020-06-17
Publication date: 2021-02-19
Anticipated expiration: 2030-06-17

Abstract

The utility model belongs to the technical field of the virus detects, a double-deck micro-fluidic chip for virus nucleic acid detects is related to. The double-layer micro-fluidic chip comprises a sample adding layer and a detection layer, wherein at least two sample adding ports are formed in the sample adding layer; the detection layer is provided with a detection cavity and a sample outlet; the detection cavity is connected with the sample adding port through a sample introduction micro-channel; the sample outlet is connected with the detection cavity through a sample outlet micro-channel. The utility model discloses a confined double-deck micro-fluidic chip detecting system, different article or reagent are added from the sample addition mouth of difference, can reduce the pollution interference that external nucleic acid brought.

Description

Double-layer micro-fluidic chip

Technical Field

The utility model belongs to the technical field of the virus detects, a double-deck micro-fluidic chip for virus nucleic acid detects is related to.

Background

Coronaviruses are a large family of viruses known to cause the common cold and more serious diseases such as Middle East Respiratory Syndrome (MERS) and Severe Acute Respiratory Syndrome (SARS). The novel coronavirus is a new strain of coronavirus that has not been previously discovered in humans. The confirmation of the novel coronavirus needs to be detected by nucleic acid and can be confirmed only if the coronavirus is positive. The existing real-time fluorescence quantitative PCR method for detecting nucleic acid has higher false negative rate, so that many infected patients can misunderstand that no infection exists, and further a wider range of human-borne infection is caused.

The existing virus detection kit has the problems of low accuracy, high false positive and false negative rate and the like in detection, and the detection process is time-consuming and cannot meet the requirement of rapid detection.

SUMMERY OF THE UTILITY MODEL

The utility model aims at providing a can be fast, simple and convenient, accurate detect the micro-fluidic chip of virus.

The utility model provides a double-layer micro-fluidic chip, which comprises a sample adding layer and a detection layer, wherein the sample adding layer is provided with at least two sample adding ports; the detection layer is provided with a detection cavity and a sample outlet; the detection cavity is connected with the sample adding port through a sample introduction micro-channel; the sample outlet is connected with the detection cavity through a sample outlet micro-channel.

As an optimized mode of the utility model, a sample introduction micro-channel is connected between each sample adding port and the detection cavity.

Further preferably, the double-layer microfluidic chip is provided with 10-50 detection units.

The utility model has the advantages that: the utility model discloses a confined double-deck micro-fluidic chip detecting system, different article or reagent are added from the sample addition mouth of difference, reduce cross contamination, can reduce the pollution interference that external nucleic acid brought. The utility model discloses a kit adopts nanotechnology and microfluidic to combine together, detects novel coronavirus (2019-nCoV), can obtain the testing result in 1 hour, and the specificity is strong, and sensitivity is high. The utility model discloses to contain detection solution, the sample that awaits measuring, cation solution, the flush fluid of RdRP gene and E gene fluorescence probe and detect in a detecting element, the process is accomplished in one step, and the reaction sequence seals completely, can reduce experiment operating procedure, saves time, has still avoided the sample pollution that the operation of uncapping probably brought simultaneously.

The probe used by the utility model is a fluorescent probe marked by CY3 at the 3' end, namely, the fluorescence emitted by the fluorescent group is absorbed by the nanometer material when in single-chain; but the reporter group can release fluorescence after the substance to be detected and the probe are subjected to hybridization reaction, and the fluorescence is detected by the fluorometer. Therefore, the utility model can be used for simple qualitative detection and quantitative detection of the specific content of the sample.

Drawings

Fig. 1 is a schematic structural diagram of a nucleic acid detection double-layer microfluidic chip according to an embodiment of the present invention;

FIG. 2 is an exploded view of FIG. 1;

fig. 3 is a structural front view of a nucleic acid detection double-layer micro-fluidic chip according to an embodiment of the present invention;

FIG. 4 is a schematic view of the structure of a sample application layer;

FIG. 5 is a schematic view of a detection layer structure.

Detailed Description

In order to facilitate understanding of the present invention, the present invention will be described in more detail with reference to the accompanying drawings and specific embodiments. Preferred embodiments of the present invention are shown in the drawings. The invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

The embodiment provided by the utility model is: a double-layer microfluidic chip is shown in figures 1-3 and comprises a sample adding layer 1 and a detection layer 2. The sample adding layer 1 is mainly used for adding a DNA probe, a sample to be detected and other reagents; the detection layer 2 is mainly used for providing a reaction container and detecting a fluorescence signal of a reactant after reaction.

As shown in fig. 1, 12 detection units are disposed on one double-layer microfluidic chip, wherein, as shown in fig. 4, each detection unit includes: the sample adding layer 1 is provided with 3 sample adding ports, namely a first sample adding port 3, a second sample adding port 5 and a third sample adding port 7. Each sample adding port is respectively connected with a sample injection micro-channel, namely a first micro-channel 4, a second micro-channel 6 and a third micro-channel 8. The 3 micro channels are independent from each other, and samples are added through respective sample adding ports, so that cross contamination can be avoided.

As shown in FIG. 5, the detection layer 2 is provided with a detection chamber 9, a sample outlet 12, and a micro flow channel 11 for sample outlet. The 3 micro-channels on the sample adding layer 1 are all connected with the detection cavity 9. The detection reagent and the sample can be injected into the detection cavity 9 through the 3 sample adding ports and the corresponding micro flow channels.

As shown in fig. 1-3, the micro-flow channel 11 of each detection unit is connected to the main flow channel 10, and the main flow channel 10 is further connected to the sample outlet 12.

The double-layer microfluidic chip of the embodiment is prepared from Polydimethylsiloxane (PDMS) and a curing agent matched with the PDMS, wherein the weight ratio of the PDMS to the curing agent is 10: 1 proportion, and the preparation.

The working principle of the double-layer micro-fluidic chip of the embodiment is as follows: firstly, a CY3 modified DNA probe (excitation wavelength is 532 nm, green light is emitted) and a nano material solution are respectively injected into a detection cavity 9 from a second sample inlet 5, a third sample inlet 7, a second flow channel 6 and a third micro flow channel 8, and the DNA probe is adsorbed by the nano material and is quenched by fluorescence.

After 5 minutes, 5 μ L of the sample to be detected is injected from the first sample inlet 3 and the first microchannel 4, and enters the detection chamber 9.

After the chip is incubated at 60 ℃ for 30 minutes, the fluorescence signal of the microfluidic chip is detected by using a spectrometer with an excitation wavelength of 532 nm.

After the detection is finished, the detection cavity 9 can be washed through the sample outlet main flow channel, the sample outlet micro flow channel 11 and the sample outlet 12, so that the purpose of repeatedly using the detection cavity 9 is achieved.

It should be noted that in this embodiment, at least 2 sample addition ports on the sample addition layer 1 are needed, one is used for adding a sample to be detected, and the other is used for adding a detection reagent, for example: DNA probe solution, nano-materials or other reagents, etc. One port can be added as a spare port.

Claims

1. A double-layer micro-fluidic chip is characterized in that: the chip comprises a sample adding layer and a detection layer, wherein at least two sample adding ports are formed in the sample adding layer; the detection layer is provided with a detection cavity and a sample outlet; the detection cavity is connected with the sample adding port through a sample introduction micro-channel; the sample outlet is connected with the detection cavity through a sample outlet micro-channel.

2. The dual-layer microfluidic chip of claim 1, wherein: and a sample introduction micro-channel is connected between each sample adding port and the detection cavity.

3. The dual-layer microfluidic chip of claim 1, wherein: each chip is provided with 10-50 detection units.