A microfluidic chip for pathogen enrichment and lysis

Abstract

The utility model provides a microfluidic chip for pathogen enrichment and lysis, and particularly relates to the technical field of virus enrichment and detection, comprising a chip lower cavity and a chip upper cavity. The microfluidic chip for pathogen enrichment and lysis has a closed lysis solution injection port, a liquid containing Escherichia coli is introduced from the sample injection port, the liquid flows into the enrichment cavity through a flow channel, the Escherichia coli cannot pass through due to the aperture barrier of a polycarbonate filter membrane, and thus is gathered in the enrichment cavity, while the liquid passes through the polycarbonate filter membrane and a porous structure, and flows out from a sample outlet hole on a sample outlet baffle, then the injection port is closed, a lysis solution is added from the lysis solution injection port, the Escherichia coli is immersed in the lysis, the chip is placed in a heating box or a heating plate for 10-15 minutes, then the lysis solution injection port and the sample outlet hole are closed, and a pipette is used to puncture the aluminum foil covering the sampling port, thereby obtaining a lysed Escherichia coli nucleic acid sample.

Description

Technical Field

[0001] The utility model relates to the technical field of virus enrichment and detection, in particular to a microfluidic chip used for enrichment and lysis of pathogenic bacteria. Background Art

[0002] Viruses are a non-cellular life form, consisting of a long nucleic acid chain and a protein shell. Viruses do not have their own metabolic machinery or enzyme system. Therefore, once they leave the host cell, they become chemical substances that have no life activities and cannot reproduce independently. Virus detection often uses bacterial detection microfluidic chips. The current pre-treatment method requires processes such as centrifugation, which is cumbersome to operate. Utility Model Content

[0003] The purpose of the utility model is to provide a microfluidic chip for pathogen enrichment and lysis, so as to solve the problem that virus detection often uses bacteria detection microfluidic chips in the above background technology, and the current pre-treatment method requires processes such as centrifugation, which is cumbersome to operate.

[0004] To achieve the above-mentioned purpose, the utility model provides the following technical scheme: comprising a chip lower chamber and a chip upper chamber, a sample liquid addition port is provided above the chip upper chamber, a lysis solution addition port is provided above the chip upper chamber, a liquid flow channel is provided on the inner side of the chip upper chamber, an enrichment chamber is provided above the chip upper chamber, a porous structure is provided on the inner side of the enrichment chamber, a sample outlet baffle is fixedly connected above the enrichment chamber, a sample outlet hole is provided through the top of the sample outlet baffle, a polycarbonate filter membrane is provided below the porous structure, a sampling port is provided below the chip lower chamber, and a sealing aluminum foil is bonded to the bottom of the sampling port of the chip lower chamber.

[0005] Preferably, the width of the flow channel is 1 mm-3 mm.

[0006] Preferably, the diameter of the pores in the porous structure is 1 mm to 2 mm.

[0007] Preferably, the diameter of the sampling port is 1 mm-3 mm.

[0008] Preferably, the thickness of the sealing aluminum foil is 0.08 mm-0.2 mm.

[0009] Compared with the prior art, the utility model has the following beneficial effects: the microfluidic chip for pathogen enrichment and lysis, the chip upper cavity, the chip lower cavity, and the sample baffle can be made of thermoplastic materials with good biocompatibility and low price, such as PP, PC, PMMA, etc., when in use, the lysis solution injection port is closed, and the liquid containing Escherichia coli is introduced from the sample injection port, and the liquid flows into the enrichment cavity through the flow channel. Due to the pore size barrier of the polycarbonate filter membrane, the Escherichia coli cannot pass through, and thus accumulate in the enrichment cavity, and the liquid passes through the polycarbonate filter membrane and the porous structure, and flows out from the sample outlet hole on the sample baffle, and then the injection port is closed, and the lysis solution injection port is added to the lysis solution injection port. coli in the lysis solution, immerse the E. coli in the lysis solution, put the chip into a heating box or heating plate for 10-15 minutes to lyse the E. coli, then seal the liquid addition port, lysis solution addition port and sample outlet port, use a pipette to puncture the aluminum foil covering the sampling port to obtain the lysed E. coli nucleic acid sample. The pore size of the polycarbonate filter membrane is about 0.2 microns. When the liquid containing E. coli flows through the filter membrane, the liquid can pass through the filter membrane, but the E. coli cannot, thereby achieving the enrichment of E. coli. The chip integrates the enrichment and lysis functions of the target bacteria, which can quickly and efficiently obtain the DNA of the target bacteria and save the pre-treatment steps of the PCR detection chip. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1 It is a structural schematic diagram of the appearance of the utility model;

[0011] Figure 2 This is a schematic diagram of the structure of the chip cavity and the liquid flow channel cooperating with each other in the utility model;

[0012] Figure 3 This is a schematic diagram of the structure in which the porous structure and the enrichment cavity cooperate with each other in the utility model.

[0013] In the figure: 1. Sample liquid addition port; 2. Lysis solution addition port; 3. Sample outlet hole; 4. Sample outlet baffle; 5. Lower cavity of chip; 6. Upper cavity of chip; 7. Liquid flow channel; 8. Porous structure; 9. Enrichment cavity; 10. Polycarbonate filter membrane; 11. Sampling port; 12. Sealing aluminum foil. DETAILED DESCRIPTION

[0014] The following will be combined with the drawings in the embodiments of the utility model to clearly and completely describe the technical solutions in the embodiments of the utility model. Obviously, the described embodiments are only part of the embodiments of the utility model, not all of the embodiments. Based on the embodiments in the utility model, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the utility model.

[0015] It should be noted that the structures, proportions, sizes, etc. illustrated in the drawings of this specification are only used to match the contents disclosed in the specification for people familiar with this technology to understand and read, and are not used to limit the conditions under which this application can be implemented. Therefore, they have no substantive technical significance. Any structural modification, change in proportional relationship or adjustment of size should still fall within the scope of the technical content disclosed in this application without affecting the effects and purposes that can be achieved by this application.

[0016] Example

[0017] See also Figure 1-3 The utility model discloses a microfluidic chip for enriching and lysing pathogens: it comprises a chip lower chamber 5 and a chip upper chamber 6, a sample liquid adding port 1 is provided above the chip upper chamber 6, a lysate adding port 2 is provided above the chip upper chamber 6, a liquid passage 7 is provided on the inner side of the chip upper chamber 6, an enrichment chamber 9 is provided above the chip upper chamber 6, a porous structure 8 is provided on the inner side of the enrichment chamber 9, a sample outlet baffle 4 is fixedly connected above the enrichment chamber 9, a sample outlet hole 3 is provided through the upper part of the sample outlet baffle 4, a polycarbonate filter membrane 10 is provided below the porous structure 8, a sampling port 11 is provided below the chip lower chamber 5, a sealing aluminum foil 12 is bonded to the bottom of the sampling port 11 of the chip lower chamber 5, the chip upper chamber 6, the chip lower chamber 5, and the sample outlet baffle 4 can be made of thermoplastic materials with good biocompatibility and low price, such as PP, PC, PMMA, etc., when in use, the lysate adding port 2 is closed, and liquid is introduced from the sample liquid adding port 1 The liquid containing Escherichia coli flows into the enrichment chamber 9 through the flow channel 7. Due to the pore size obstruction of the polycarbonate filter membrane 10, the Escherichia coli cannot pass through and thus accumulate in the enrichment chamber 9. The liquid passes through the polycarbonate filter membrane 10 and the porous structure 8 and flows out from the sample outlet 3 on the sample outlet baffle 4. Then the liquid addition port 1 is closed, and the lysis solution is added from the lysis solution addition port 2 to immerse the Escherichia coli 13 in the lysis. The chip is placed in a heating box or on a heating plate for 10-15 minutes to lyse the Escherichia coli. Then the liquid addition port 1, the lysis solution addition port 2 and the sample outlet 3 are closed, and the aluminum foil 12 covering the sampling port 11 is punctured with a pipette to obtain the lysed Escherichia coli nucleic acid sample. The pore size of the polycarbonate filter membrane 10 is about 0.2 microns. When the liquid containing Escherichia coli flows through the filter membrane 10, the liquid can pass through the filter membrane, but the Escherichia coli cannot pass through, thereby achieving the enrichment of the Escherichia coli.

[0018] Furthermore, the width of the flow channel 7 is 1 mm-3 mm.

[0019] Furthermore, the diameter of the small holes in the porous structure 8 is 1 mm to 2 mm.

[0020] Furthermore, the diameter of the sampling port 11 is 1 mm-3 mm.

[0021] Furthermore, the thickness of the sealing aluminum foil 12 is 0.08 mm-0.2 mm.

[0022] Working principle: Escherichia coli is injected into the second sample mixture inlet 7 through a pressure pump or an injection pump, and coliform bacteria is injected into the first sample mixture inlet 6. Escherichia coli is mixed with the oil phase and discharged through the first droplet outlet 8 to the first droplet storage chamber 2. Coliform bacteria is mixed with the oil phase and discharged through the second droplet outlet 9 to the second droplet storage chamber 3, thereby completing the simultaneous detection of Escherichia coli and coliform bacteria. The structure of the commercial cell counting plate 15 itself is a semicircular hole, and then the droplets are sucked into the storage chamber through the siphon action to achieve the effect of non-interference. At the same time, the cell counting plate can be directly placed in a cell counter to count and read the results. The droplet-based detection method also greatly shortens the detection time of Escherichia coli and coliform bacteria from the traditional 24 hours to 3 hours, greatly saving time and labor costs. Because the detection is carried out in microdroplets, the sample volume is reduced. Under the same reaction time, the concentration of the generated substance to be detected is higher and can be detected in 3 hours.

[0023] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the implementation rules without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A microfluidic chip for pathogen enrichment and lysis, comprising a chip lower chamber (5) and a chip upper chamber (6), characterized in that: A sample liquid injection port (1) is provided above the upper chamber (6) of the chip, a lysate injection port (2) is provided above the upper chamber (6), a liquid flow channel (7) is provided inside the upper chamber (6), an enrichment chamber (9) is provided above the upper chamber (6), a porous structure (8) is provided inside the enrichment chamber (9), a sample outlet baffle (4) is fixedly connected above the enrichment chamber (9), a sample outlet hole (3) is provided through the upper portion of the sample outlet baffle (4), a polycarbonate filter membrane (10) is provided below the porous structure (8), a sampling port (11) is provided below the lower chamber (5) of the chip, and a sealing aluminum foil (12) is bonded to the bottom of the sampling port (11) of the lower chamber (5) of the chip.

2. A microfluidic chip for pathogen enrichment and lysis according to claim 1, characterized in that: The width of the flow channel (7) is 1 mm-3 mm.

3. A microfluidic chip for pathogen enrichment and lysis according to claim 1, characterized in that: The diameter of the small holes on the porous structure (8) is 1 mm to 2 mm.

4. A microfluidic chip for pathogen enrichment and lysis according to claim 1, characterized in that: The diameter of the sampling port (11) is 1 mm-3 mm.

5. A microfluidic chip for pathogen enrichment and lysis according to claim 1, characterized in that: The thickness of the sealing aluminum foil (12) is 0.08 mm-0.2 mm.

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