CN220126240U - Chip bottom shell of micro-fluidic chip and micro-fluidic chip - Google Patents

Abstract

The utility model discloses a chip bottom shell of a microfluidic chip and the microfluidic chip, which relate to the technical field of nucleic acid detection, and the chip bottom shell and the microfluidic chip comprise: a substrate; the extraction cavity is arranged on the substrate; and the detection reagent storage module is arranged on the substrate and comprises a plurality of columnar storage cavities which can be matched with the piston covers and are communicated with the extraction cavity, the axes of the columnar storage cavities are parallel to the surface of the substrate, the piston covers of the columnar storage cavities are matched with openings on the thickness surface of the substrate, and the thickness surface of the substrate is perpendicular to the surface of the substrate. The chip bottom shell of the microfluidic chip and the microfluidic chip are reasonable and simple in structure, convenient to manufacture, high in space utilization rate in stacking and convenient to transport and store.

Description

Chip bottom shell of micro-fluidic chip and micro-fluidic chip

Technical Field

The utility model relates to the technical field of nucleic acid detection, in particular to a chip bottom shell of a microfluidic chip and the microfluidic chip.

Background

The microfluidic chip technology integrates basic operation units of sample preparation, reaction, separation, detection and the like in biological, chemical and medical analysis processes onto a micron-scale chip, and automatically completes the whole analysis process. The microfluidic chip has the characteristics of small volume, low components, high efficiency, automation, integration and the like, and is a detection chip which integrates nucleic acid extraction, purification, elution and detection into a whole by using a complex fluid operation system and a functional unit module on a chip with a small area. However, if the extraction and amplification of nucleic acid are to be realized on the microfluidic chip, the microfluidic chip mostly comprises complex liquid paths and control valve designs, so that the manufacturing cost of the microfluidic chip is high, the whole volume is difficult to further reduce, and the transportation and storage costs are high.

Disclosure of Invention

The utility model aims to overcome the technical problems and provide the chip bottom shell of the micro-fluidic chip and the micro-fluidic chip, which have reasonable and simple structures, are convenient to manufacture, have high space utilization rate in stacking and are convenient to transport and store.

In order to achieve the above object, the present utility model provides a chip bottom case of a microfluidic chip, the chip bottom case comprising:

a substrate;

the extraction cavity is arranged on the substrate; and

the detection reagent storage module is arranged on the substrate and comprises a plurality of columnar storage cavities which can be matched with the piston cover and are communicated with the extraction cavity, the columnar storage cavities are longitudinally arranged side by side along the plate surface of the substrate, the axes of the columnar storage cavities are transversely arranged along the plate surface of the substrate, the piston cover matched openings of the columnar storage cavities are arranged on the thickness surface of the substrate, and the thickness surface of the substrate is perpendicular to the plate surface of the substrate.

Further, the chip bottom shell can be an integrated part, and the chip bottom shell further comprises a sample adding cavity, a waste liquid cavity and a plurality of amplification cavities which are communicated with the extraction cavity respectively, wherein the substrate comprises a first plate surface and a second plate surface which are oppositely arranged along the thickness direction of the substrate, the sample adding cavity, the extraction cavity, the detection reagent storage module and the waste liquid cavity are all arranged on the first plate surface and are used for being communicated with a micro-channel between the cavities, and the amplification cavities are arranged on the second plate surface.

Further, the sample adding cavity and the extracting cavity can be arranged side by side with the detection reagent storage module along the transverse direction of the plate surface of the substrate.

Optionally, the sample adding cavity, the extracting cavity and the waste liquid cavity can be in an open top shape, the cavity bottom wall is arranged on the first plate surface, the cavity peripheral walls of the sample adding cavity, the extracting cavity and the waste liquid cavity are all convex and formed on the first plate surface, and the amplifying cavities are in an open shape and concave and formed on the second plate surface.

In some embodiments, the extraction chamber and the detection reagent storage module may be arranged adjacently and share a portion of the chamber wall.

Further, the plurality of columnar storage chambers may include:

the first storage cavities are communicated with the extraction cavity through first flow channels, the first flow channels comprise first main flow channels and first branch flow channels, first ends of the first branch flow channels are respectively communicated with the first storage cavities one by one, and the first main flow channels are communicated between the extraction cavity and second ends of the first branch flow channels; and

the second storage cavity is communicated with the extraction cavity through a second flow channel.

The plurality of first branch flow passages are provided with first branch flow passage cutting portions for cutting off the flow passages, and the second flow passage is provided with second flow passage cutting portions for cutting off the flow passages.

The utility model also provides a microfluidic chip, which comprises the chip bottom shell.

Further, the detection reagent storage module may be provided with a vesicle extraction reagent, a vesicle washing liquid reagent and a vesicle eluting liquid reagent, the amplification cavity is provided with a nucleic acid amplification freeze-drying detection reagent ball, and the microfluidic chip may further include:

the first coating is welded on one side of the bottom shell of the chip and is used for sealing the waste liquid cavity and the extraction cavity;

the second coating is welded on the other side of the bottom shell of the chip and is used for sealing the micro-flow channel and the plurality of amplification cavities of the second plate surface;

the sample adding sealing plug is used for sealing a sample adding port of the sample adding cavity and can perform piston movement in the sample adding cavity;

the upper cover is arranged on the bottom shell of the chip and is provided with a sample adding hole and an ultrasonic hole, the sample adding hole is aligned with the sample adding cavity, and the ultrasonic hole is aligned with the extraction cavity; and

the piston covers are sealed in the columnar storage cavities in a one-to-one fit manner.

Further, one end of the piston cover may be a plunger end, and the other end may be provided with a piston rod connecting portion capable of being connected to the piston rod.

The chip bottom shell and the microfluidic chip comprise a substrate, an extraction cavity and a detection reagent storage module, wherein the extraction cavity and the detection reagent storage module are arranged on the substrate, the detection reagent storage module comprises a plurality of columnar storage cavities which can be matched with a piston cover and are communicated with the extraction cavity, the columnar storage cavities are longitudinally arranged side by side along the surface of the substrate, the axes of the columnar storage cavities are transversely arranged along the surface of the substrate, the piston cover matched openings of the columnar storage cavities are arranged on the thickness surface of the substrate, and the thickness surface of the substrate is vertical to the surface of the substrate, so that the microfluidic chip is in a flat plate shape, has small thickness, and has high space utilization rate during storage or stacking and is convenient to transport and store. The chip bottom shell and the microfluidic chip of the utility model have reasonable and simple structures and are convenient to manufacture. In addition, the columnar storage cavity is used for storing reactive reagent vesicles, the piston driving motor is driven, the vesicles in the columnar storage cavity are extruded by the plunger, after the vesicles are pressed and broken, the reactive reagents in the vesicles enter the extraction cavity from the flow channel, namely, the reactive reagents in the vesicles in the corresponding storage cavities are selectively led in by the corresponding piston driving motor to be mixed in the extraction cavity by a sample, so that sample processing is realized in a closed space, the risk of pollution during transfer of the reactive reagents is effectively avoided, and the processing flow is simplified.

Additional features and advantages of the utility model will be set forth in the detailed description which follows.

Drawings

The accompanying drawings are included to provide a further understanding of the utility model, and are incorporated in and constitute a part of this specification, illustrate the utility model and together with the description serve to explain, without limitation, the utility model. In the drawings:

fig. 1 illustrates an exploded view of the mounting of a microfluidic chip according to one embodiment of the present utility model;

FIG. 2 is a perspective view of the bottom case of the chip of FIG. 1;

FIG. 3 shows a first plate of the bottom case of FIG. 2;

FIG. 4 shows a second plate of the bottom case of FIG. 2;

FIG. 5 is a perspective view of the loaded sealing plug of FIG. 1;

fig. 6 is a perspective view of the upper cover of fig. 1.

Description of the reference numerals

100. Chip bottom case 101 substrate

1011 first panel 1012 second panel

1013 substrate thickness surface 102 sample adding cavity

1021 sample adding cavity liquid outlet hole 1022 sample adding cavity liquid outlet micro-channel

103. Extraction chamber 1031 extraction chamber external vent

1032 extraction chamber vent flow channel 1033 extraction chamber vent

1034 extraction chamber drain 1035 eluent discharge port

1036 extraction Chamber liquid feed hole 1037 first reagent liquid feed hole

1038 second reagent inlet 1071 wax outlet

104. Detection reagent storage module 1041 piston cap mating opening

1042 first storage Chamber 1043 second storage Chamber

1044 first flow path 1045 second flow path

1046 first branch flow path cut-off portion 1047 second branch flow path cut-off portion

105. Waste liquid chamber 1051 waste liquid chamber outer vent

1052 waste liquid cavity exhaust runner 1053 waste liquid cavity exhaust hole

1054 waste liquid cavity liquid inlet 1055 waste liquid cavity micro-channel

1056 waste liquid chamber micro-channel cutting part 106 amplification chamber

1061 amplification chamber micro-channel 10611 amplification chamber main channel

10612 amplification chamber branch flow channel 1062 amplification chamber micro flow channel cutting part

1063 amplification chamber exhaust micro-channel 1064 amplification chamber exhaust hole

1065 amplification chamber branch channel cutting part 107 wax valve cavity

108. Positioning column 200 sample-adding sealing plug

300. Upper cover 301 sample-adding hole

302. Ultrasonic hole 400 piston cover

303. Positioning pin

Detailed Description

The following describes specific embodiments of the present utility model in detail with reference to the drawings. It should be understood that the detailed description and specific examples, while indicating and illustrating the utility model, are not intended to limit the utility model.

The microfluidic chip in the prior art mostly comprises complicated liquid path and control valve designs, and has higher design and material use requirements, so that the manufacturing cost of the microfluidic chip is higher, and the volume is difficult to further miniaturize.

In view of this, the present utility model provides a chip bottom case using a microfluidic chip, as shown in fig. 1 to 4, the chip bottom case 100 includes a substrate 101, a sample loading cavity 102, an extraction cavity 103, a detection reagent storage module 104, a waste liquid cavity 105, and a plurality of amplification cavities 106, where the sample loading cavity 102, the extraction cavity 103, the detection reagent storage module 104, the waste liquid cavity 105, and the plurality of amplification cavities 106 are all disposed on the substrate 101, and the sample loading cavity 102, the detection reagent storage module 104, the waste liquid cavity 105, and the plurality of amplification cavities 106 are respectively communicated with the extraction cavity 103 through micro channels on the substrate 101, and the chip bottom case 100 is an integral molding. Namely, as shown in fig. 1 to 4, the sample adding cavity 102, the detection reagent storage module 104, the waste liquid cavity 105 and the amplification cavities 106 are respectively communicated with the extracting cavity 103 through micro-channels on the substrate 101, the extracting cavity 103 is directly communicated with other cavities through the micro-channels, the flow channel arrangement is simple, a control valve with a complex structure is not needed, the structure is simple and reasonable, and the requirements on design and material use are not high. And the chip bottom shell 100 is an integral molding piece, so that the manufacturing cost is low.

The detection reagent storage module 104 includes a plurality of columnar storage chambers capable of being mated with the piston cover 400 and communicating with the extraction chamber 103, axes of the plurality of columnar storage chambers are parallel to a plate surface of the substrate 101, the plurality of columnar storage chambers are longitudinally arranged side by side along the plate surface of the substrate 101, axes of the plurality of columnar storage chambers are transversely arranged along the plate surface of the substrate 101, piston cover mating openings 1041 of the plurality of columnar storage chambers are provided on a substrate thickness surface 1013 of the substrate 101, and the substrate thickness surface 1013 is perpendicular to the plate surface of the substrate 101. The axes of the columnar storage cavities are transversely arranged along the plate surface of the substrate 101, so that the structure is reasonable and simple, the manufacture is convenient, the space utilization rate in stacking is high, and the transportation and storage are convenient.

In addition, the columnar storage cavity is used for storing reactive reagent vesicles, the piston driving motor is driven, the vesicles in the columnar storage cavity are extruded by the plunger, after the vesicles are pressed and broken, the reactive reagents in the vesicles enter the extraction cavity from the flow channel, namely, the reactive reagents in the vesicles in the corresponding storage cavities are selectively led in by the corresponding piston driving motor to be mixed in the extraction cavity by a sample, so that sample processing is realized in a closed space, the risk of pollution during transfer of the reactive reagents is effectively avoided, and the processing flow is simplified.

Alternatively, as shown in fig. 4 and 5, the substrate 101 may include a first plate 1011 and a second plate 1012 opposite to each other along the thickness direction of the substrate, where the sample loading chamber 102, the extraction chamber 103, the detection reagent storage module 104, and the waste liquid chamber 105 are all disposed on the first plate 1011, and the micro flow channel and the plurality of amplification chambers 106 are disposed on the second plate 1012. In this way, the arrangement of the chambers and the arrangement of the channels on the chip bottom shell 100 are more reasonable, which is more beneficial to the further miniaturization of the chip bottom shell 100.

In addition, along the longitudinal direction of the plate surface of the substrate 101, the sample loading chamber 102, the extraction chamber 103, the waste liquid chamber 105 and the amplification chamber 106 are sequentially arranged from top to bottom, the plurality of amplification chambers 106 are positioned under the longitudinal direction of the extraction chamber 103, and the sample loading chamber 102 and the extraction chamber 103 are arranged side by side with the detection reagent storage module 104 along the transverse direction of the plate surface of the substrate 101. Therefore, the structural arrangement of the microfluidic chip is simpler and more reasonable, the transfer of liquid between the chambers can be realized by means of gravity, the operation is convenient, and the usability is good.

Further, as shown in fig. 3 and 4, the loading chamber 102 and the extraction chamber 103 are arranged side by side with the detection reagent storage module 104 in the lateral direction of the plate surface of the substrate 101. Thus, the transverse ends of the plurality of columnar storage cavities are provided with the piston cover matching openings 1041 and are located on the substrate thickness surface 1013 of the substrate 101, and the transverse ends of the plurality of columnar storage cavities are communicated with the extraction cavity 103 through the short flow channels, so that the flow channel arrangement between the plurality of columnar storage cavities and the extraction cavity 103 is more concise and reasonable.

Further, the plurality of columnar storage chambers includes a plurality of first storage chambers 1042 and second storage chambers 1043. The plurality of first storage cavities 1042 are communicated with the extraction cavity 103 through a first flow channel 1044 and a first reagent inlet 1037, the first flow channel comprises a first main flow channel and a plurality of first branch flow channels, first ends of the plurality of first branch flow channels are respectively communicated with the plurality of first storage cavities 1042 one by one, and the first main flow channel is communicated between the extraction cavity 103 and second ends of the plurality of first branch flow channels. The second storage chamber 1043 communicates with the extraction chamber 103 through the second flow passage 1045 and the second reagent feed 1038. The plurality of first branch flow passages are provided with first branch flow passage cutting portions 1046 for cutting off flow passages, and the second flow passage 1045 is provided with second flow passage cutting portions 1047 for cutting off flow passages.

Alternatively, the piston cover 400 has a cylinder end at one end and a piston rod connecting portion capable of connecting with a piston rod at the other end.

In some embodiments, the first storage cavities 1042 may set a certain number of reserved slots according to the number actually required to adapt to different kinds of detection requirements. As shown in fig. 1-5, the detection reagent storage module 104 includes five first storage chambers 1042 and one second storage chamber 1043. The reagents required by nucleic acid extraction are pre-loaded into the first storage cavity 1042 in a vesicle form, eluent is placed into the second storage cavity 1043, then the piston cover 400 is plugged into the piston cover matching openings 1041 of the plurality of columnar storage cavities, and a tearable dustproof paper film is stuck on the surface to finish the packaging. After the sealing, the liquid reagent is sealed by a flow path membrane valve formed by the piston cover 400, the first and second branch flow path cut-off portions 1046 and 1047. In the experiment, the testing instrument pushes a specific plunger through a screw motor, extrudes the vesicle to break, and the liquid reagent rushes the membrane breaking valve to flow into the extraction cavity 103.

Further, as shown in fig. 1 to 4, the sample loading chamber 102, the extraction chamber 103 and the waste liquid chamber 105 may be opened at the top, and the bottom walls of the sample loading chamber 102, the extraction chamber 103 and the waste liquid chamber 105 are disposed on the first plate surface 1011, the peripheral walls of the sample loading chamber 102, the extraction chamber 103 and the waste liquid chamber 105 are all formed on the first plate surface 1011 in a protruding manner, and the amplification chambers 106 are opened and concave on the second plate surface 1012. Thus, the arrangement of the chambers and the arrangement of the flow channels on the chip bottom shell 100 are more reasonable, the manufacturing materials and the weight of the chip bottom shell 100 can be reduced, the manufacturing cost is low, the further miniaturization and the light weight arrangement of the chip bottom shell 100 are also more facilitated, the space utilization rate in stacking is high, and the transportation and the storage are facilitated.

Optionally, the extraction chamber 103 is arranged adjacent to the detection reagent storage module 104 and shares part of the chamber wall. In this way, the arrangement of the chambers and the arrangement of the flow channels on the chip bottom shell 100 are more reasonable, the manufacturing materials and the weight of the chip bottom shell 100 can be reduced, the manufacturing cost is low, and the further miniaturization and the light-weight arrangement of the chip bottom shell 100 are more facilitated.

The chip bottom case 100 and the microfluidic chip 1000 of the present utility model include a plurality of amplification chambers 106, and as shown in fig. 1, 3 to 5, the chip bottom case 100 and the microfluidic chip 1000 of the present utility model include 8 amplification chambers 106, but the present utility model is not limited thereto, and the number of amplification chambers 106 may be more, for example, 10, 12, etc. based on reasonable flow channels and chamber layouts of the present utility model. The chip bottom shell 100 and the microfluidic chip 1000 can be pre-embedded with a liquid reagent for nucleic acid extraction by a magnetic bead method and a freeze-dried reagent for nucleic acid amplification, one consumable material is used, one sample is added, the operation time of a person is less than one minute, and the fluorescent PCR detection of up to 32 targets can be realized by matching with an instrument to finish the nucleic acid extraction and amplification flow of a sample inlet and a sample outlet, so that the detection efficiency is high.

The loading chamber 102 is provided with a loading port, and the loading chamber 102 is located longitudinally above the extraction chamber 103 along the plate surface of the substrate 100. The extraction chamber 103 is communicated with an extraction chamber exhaust passage capable of exhausting outwards, and the extraction chamber exhaust passage comprises an extraction chamber external exhaust hole 1031, an extraction chamber exhaust runner 1032 and an extraction chamber internal exhaust hole 1033. The extraction chamber exhaust flow channel 1032 is formed on the second plate surface 1012 in a concave manner, and the extraction chamber external exhaust hole 1031 and the extraction chamber internal exhaust hole 1033 are both provided on the substrate 101 in a penetrating manner in the substrate thickness direction. The extraction chamber outside air outlet 1031 is located longitudinally above the extraction chamber 103 along the plate surface longitudinal direction of the substrate 101. The extraction chamber inner vent 1033 is provided to penetrate the bottom wall of the chamber body above the longitudinal direction of the extraction chamber 103 in the thickness direction of the substrate, and the extraction chamber 103, the extraction chamber inner vent 1033, the extraction chamber vent runner 1032, and the extraction chamber outer vent 1031 are sequentially communicated.

Thus, since the extraction cavity 103 is provided with the exhaust channel, after the sample liquid is added into the sample adding cavity 102, the sample liquid can be injected into the extraction cavity 103 by utilizing the gravity and the forward extrusion action of the sample to be added through the plug-in sample adding sealing plug 200, and no negative pressure is required to be provided by other cavities. The microfluidic chip disclosed by the utility model has the advantages of simple structure, low manufacturing cost, capability of using one consumable, one-time sample adding, less than one minute of personnel operation time, convenience in operation and good user experience.

In addition, the waste cavity 105 may be connected to a waste cavity exhaust channel capable of exhausting outwards, and the waste cavity exhaust channel includes a waste cavity external exhaust hole 1051, a waste cavity exhaust flow channel 1052, and a waste cavity internal exhaust hole 1053. The waste liquid chamber external vent hole 1051 is provided penetrating the substrate 101 in the substrate thickness direction along the longitudinal direction of the plate surface of the substrate 101 and is located longitudinally above the waste liquid chamber 105. The waste liquid chamber inner vent 1053 penetrates through the substrate 101 and the bottom wall of the chamber body of the waste liquid chamber 105 along the thickness direction of the substrate, the waste liquid chamber vent channel 1052 is concavely formed on the second plate surface 1012, and the waste liquid chamber 105, the waste liquid chamber inner vent 1053, the waste liquid chamber vent channel 1052 and the waste liquid chamber outer vent 1051 are sequentially communicated. Therefore, the microfluidic chip of the utility model can transfer liquid and liquid between the extraction cavity 103 and the waste liquid cavity 105 by gravity without providing additional power, is convenient for users to use, and has a simple and reasonable structure by integrally forming the exhaust channel on the chip bottom shell 100.

Further, the second plate 1012 may be concavely formed with a waste liquid chamber micro flow channel 1055, the extraction chamber 103 is provided with an extraction chamber liquid drain hole 1034 penetrating the substrate 101 and the bottom wall of the cavity of the extraction chamber 103 along the thickness direction of the substrate, the waste liquid chamber 105 is provided with a waste liquid chamber liquid inlet hole 1054 penetrating the substrate 101 and the bottom wall of the cavity of the waste liquid chamber 105 along the thickness direction of the substrate, and the extraction chamber 103 is communicated with the waste liquid chamber 105 sequentially through the extraction chamber liquid drain hole 1034, the waste liquid chamber micro flow channel 1055 and the waste liquid chamber liquid inlet hole 1054.

Further, the second plate 1012 is concave downward to form an amplification chamber microchannel, the amplification chambers 106 are communicated with the extraction chamber 103 through the amplification chamber microchannel, and an extraction chamber microchannel cutting part capable of forming a membrane valve is formed on the extraction chamber microchannel.

Alternatively, a plurality of amplification chambers 106 may be arranged side by side in the lateral direction of the plate surface of the substrate 101.

In some embodiments, the extraction chamber 103 may be in communication with the waste chamber 105 through a waste chamber microchannel 1055, the waste chamber microchannel 1055 is provided with a waste chamber valve capable of controlling the flow passage to be opened or closed, the extraction chamber 103 is in communication with the plurality of amplification chambers 106 through an amplification chamber microchannel 1061, and the amplification chamber microchannel 1061 is provided with an amplification chamber valve capable of controlling the flow passage to be opened or closed. The waste liquid chamber micro flow channel 1055 and the amplification chamber micro flow channel 1061 are both concaved downwards on the second plate surface 1012, a waste liquid chamber micro flow channel cutting part 1056 for cutting off the flow channel is integrally arranged on the waste liquid chamber micro flow channel 1055, the waste liquid chamber micro flow channel cutting part 1056 and the second coating form a waste liquid chamber valve, an amplification chamber micro flow channel cutting part 1062 for cutting off the flow channel is integrally arranged on the amplification chamber micro flow channel 1061, and the amplification chamber micro flow channel cutting part 1062 and the second coating form an amplification chamber valve.

Further, the multiple amplification chambers 106 are communicated with the extraction chamber 103 through the amplification chamber micro flow channel 1061, the amplification chamber micro flow channel 1061 may include an amplification chamber main flow channel 10611 and multiple amplification chamber sub flow channels 10612, the first ends of the amplification chamber main flow channels 10611 are communicated with the extraction chamber 103, the first ends of the multiple amplification chamber sub flow channels 10612 are respectively communicated with the multiple amplification chambers 106 one by one, the second ends of the multiple amplification chamber sub flow channels 10612 are communicated with the second ends of the amplification chamber main flow channels 10611, and the amplification chamber micro flow channel cut-off portion 1062 is arranged on the amplification chamber main flow channel 10611. Each amplification chamber branch channel 10612 is provided with an amplification chamber branch channel cutting portion 1065 for cutting off the channel, and each amplification chamber branch channel cutting portion 1065 and the second coating film form an amplification chamber branch channel valve.

Still further, the extraction chamber 103 may be provided with an eluent discharge hole 1035 penetrating the bottom wall of the chamber and the substrate 101 in the thickness direction of the substrate, the eluent discharge hole 1035 being located at a longitudinally lower portion of the extraction chamber 103, the amplification chamber main channel 10611 being communicated with the extraction chamber 103 through the eluent discharge hole 1035, and the amplification chamber micro channel cut-off portion 1062 being provided between the amplification chamber main channel 10611 and the eluent discharge hole 1035.

Optionally, the amplification chambers 106 are communicated with an amplification chamber exhaust channel capable of exhausting outwards, and the amplification chamber exhaust channel comprises a plurality of amplification chamber exhaust micro-channels 1063 and a plurality of amplification chamber exhaust holes 1064 arranged on the amplification chamber exhaust micro-channels 1063. Along the longitudinal direction of the plate surface of the substrate, a plurality of amplification cavity exhaust holes 1064 are formed in the substrate 101 in a penetrating manner along the thickness direction of the substrate and are positioned above the longitudinal direction of the plurality of amplification cavities 106, and the plurality of amplification cavities 106, the plurality of amplification cavity exhaust micro-channels 1063 and the plurality of amplification cavity exhaust holes 1064 are sequentially communicated in a one-to-one correspondence manner. The microfluidic chip 1000 may further include a waterproof and breathable film covering the amplification chamber exhaust hole 1064 located on the first plate surface 1011.

In some embodiments, the waste liquid chamber micro flow channel 1055 and the amplification chamber micro flow channel 1061 are both concaved on the second plate surface 1012, the waste liquid chamber micro flow channel 1055 is integrally provided with a waste liquid chamber micro flow channel cutting part 1056 for cutting off the flow channel, the waste liquid chamber micro flow channel cutting part 1056 and the second coating form a waste liquid chamber valve, the amplification chamber micro flow channel 1061 is integrally provided with an amplification chamber micro flow channel cutting part 1062 for cutting off the flow channel, the amplification chamber micro flow channel cutting part 1062 and the second coating form an amplification chamber valve, and the waste liquid chamber valve and the amplification chamber valve form a normally open membrane valve. Wherein, when the microfluidic chip 1000 is placed at the detection position, the waste liquid chamber valve and the amplification chamber valve can be respectively pressed by the cutting-portion driving motor to close the second cover film on the waste liquid chamber flow channel cutting portion 1056 and the amplification chamber micro flow channel cutting portion 1062.

Optionally, the plurality of amplification chambers 106 may be communicated with an amplification chamber exhaust channel capable of exhausting outwards, the amplification chamber exhaust channel is provided with a wax valve chamber 107, the plurality of wax valve chambers 107 are integrally formed on the substrate 101, the wax valve chamber 107 is in an open top shape, and the bottom wall of the chamber is arranged on the first plate surface 1011. The waste liquid chamber 105 is located between the extraction chamber 103 and the wax valve chamber 107 and extends from one end to the other end in the widthwise direction of the plate surface of the substrate 101. The plurality of wax valve chambers 107 and the plurality of amplification chambers 106 are arranged side by side in the lateral direction of the plate surface of the substrate 101, respectively.

The middle part of the flow channel of each amplification cavity branch flow channel 10612 and the end of the flow channel of each amplification cavity exhaust micro flow channel 1063 are respectively communicated with a wax valve cavity 107, paraffin is filled in the wax valve cavity 107, and the extraction cavity 103, the plurality of wax valve cavities 107, the plurality of amplification cavity exhaust holes 1064 and the plurality of amplification cavities 106 are sequentially arranged from top to bottom along the longitudinal direction of the plate surface of the substrate 101. The flow channel end of the amplification chamber exhaust micro flow channel 1063 extends longitudinally upward along the plate surface of the substrate 101, and the amplification chamber exhaust vent 1064 is located on the flow channel between the flow channel end of the amplification chamber exhaust micro flow channel 1063 and the amplification chamber 106.

Nucleic acid amplification freeze-drying detection reagent balls containing different primers are pre-buried in the amplification cavity 106, because the front surface of the amplification cavity exhaust hole 1064 is covered with a waterproof and breathable film, when the wax valve cavity 107 is not heated, solid paraffin in the wax valve cavity 107 can respectively block a hole communicated with the wax valve cavity 107 at the end of a runner of the amplification cavity exhaust micro runner 1063 and a wax outlet 1071 communicated with the wax valve cavity 107 in the middle of a runner of the amplification cavity branch runner 10612, and the inside freeze-drying balls of the amplification cavity 106 are in a relatively airtight state with the outside. After the nucleic acid-containing eluent is injected from the bottom of the amplification cavity 106 to fill the amplification cavity 106, a heating module on the instrument melts paraffin in the paraffin valve cavity 107, and the melted paraffin correspondingly flows into the channels along the amplification cavity exhaust micro-channel 1063 and the amplification cavity branch channel 10612 so as to seal the amplification cavity exhaust hole 1064, the amplification cavity exhaust micro-channel 1063 and the amplification cavity branch channel 10612, thereby forming a complete seal of the amplification cavity 106.

Optionally, the waste chamber 105 is located longitudinally above the wax valve chamber 107, the waste chamber 105 being arranged adjacent to the wax valve chamber 107 and sharing part of the chamber wall.

The utility model also provides a microfluidic chip, which comprises the chip bottom shell 100.

Further, the detection reagent storage module 104 is provided with an extraction reagent, a washing liquid reagent and an eluting liquid reagent, and the amplification chamber 106 is provided with a nucleic acid amplification freeze-dried detection reagent ball. As shown in fig. 1, 5 and 6, the microfluidic chip further includes a first coating, a second coating, a sample-loading sealing plug 200, an upper cover 300, and a plurality of piston covers 400. The first coating is welded on one side of the chip bottom shell 100 and is used for sealing the extraction cavity 103, the waste liquid cavity 105 and the wax valve cavity 107, and the second coating is welded on the other side of the chip bottom shell 100 and is used for sealing the micro flow channel of the second plate surface 1012 and the plurality of amplification cavities 106. The sample loading sealing plug 200 is used for sealing a sample loading port and can perform a piston movement in the sample loading cavity 101, the upper cover 300 is arranged on the chip bottom shell 100 in a covering manner and is provided with a sample loading hole 301 and an ultrasonic hole 302, the sample loading hole 301 is arranged in alignment with the sample loading cavity 102, and the ultrasonic hole 302 is arranged in alignment with the extraction cavity 103. The plurality of piston caps 400 are capped in a one-to-one fit in the plurality of columnar storage cavities.

The sample-adding sealing plug 200 and the piston cover 400 may be made of elastic rubber or silica gel, the upper cover 300 is a plastic housing, and the upper surface may be attached with or printed with information such as chip identification mark. The chip bottom shell 100 is a colorless transparent plastic chip bottom shell, and transparent pp films are coated on two sides of the chip bottom shell through ultrasonic welding, namely, the first coating film and the second coating film are transparent pp films coated through ultrasonic welding. The upper cover 300 and the bottom chip housing 100 may be connected and fixed by the positioning posts 108 and the positioning pins 303.

The utility model also provides a nucleic acid extraction fluorescence PCR detection system, and the nucleic acid extraction fluorescence PCR detection system is applied to the microfluidic chip.

The application of the microfluidic chip to the nucleic acid extraction fluorescence PCR detection system uses a microfluidic chip to detect by matching with an instrument, realizes multiple fluorescence PCR detection with the personnel operation time less than one minute, sample in and out, and has simple operation and high detection efficiency.

The nucleic acid extraction fluorescence PCR detection method comprises the following steps:

step S1, providing a microfluidic chip 1000;

s2, injecting a sample liquid to be detected into the extraction cavity 103 through the sample adding cavity 102;

step S3, injecting the extraction reagent, the washing liquid reagent and the eluting liquid reagent in the detection reagent storage module 104 into the extraction cavity 103 in batches to react with the sample to be detected, respectively discharging the reacted waste liquid into the waste liquid cavity 105, and injecting the obtained eluting liquid containing nucleic acid into the amplification cavities 106;

and S4, starting the fluorescent PCR module to detect the mixed solution in the amplification chambers 106.

Optionally, step S2 may specifically include:

placing the microfluidic chip 1000 flat so that the sample loading port faces upwards, and loading a sample to be tested from the sample loading port;

the sample-adding sealing plug 200 is plugged into the sample-adding port, and the microfluidic chip 1000 is vertically placed at the detection position along the longitudinal direction of the substrate, i.e. the amplification chamber 106 is vertically placed in the detection instrument in the downward direction, so that the sample liquid to be detected in the sample-adding chamber 102 flows down into the extraction chamber 103. That is, a user uses a quantitative sample adding tool to add sample liquid into the sample adding cavity 102, plugs the sample adding sealing plug 200, vertically places the microfluidic chip 1000, and the liquid flows into the extracting cavity 103 along the vertical direction of the liquid outlet micro-channel 1022 of the sample adding cavity under the action of gravity and extrusion.

Further, step S3 may specifically include:

starting a piston driving motor to push a piston of a columnar storage cavity storing the extraction reagent to move, and injecting the extraction reagent into the extraction cavity 103;

starting an ultrasonic instrument, and performing ultrasonic pyrolysis on the sample liquid to be detected in the extraction cavity 103;

closing the ultrasonic instrument, matching the magnetic attraction instrument with the extraction cavity 103 and discharging the waste liquid in the extraction cavity 103 into the waste liquid cavity 105;

starting a piston driving motor to push a piston cover 400 of a columnar storage cavity storing a washing liquid reagent to move, and injecting the washing liquid reagent into an extraction cavity;

removing the magnetic attraction instrument and starting the ultrasonic instrument to wash the substances in the extraction cavity 103;

closing the ultrasonic instrument, matching the magnetic attraction instrument with the extraction cavity 103 and discharging the waste liquid in the extraction cavity 103 into the waste liquid cavity 105;

starting a piston driving motor to push a piston cover 400 of a columnar storage cavity storing the eluting liquid reagent to move, and injecting the eluting liquid reagent into an extraction cavity 103;

removing the magnetic attraction instrument and starting the ultrasonic instrument to elute the substances in the extraction cavity 103;

the ultrasound instrument is turned off, the magnetic attraction instrument is mated with the extraction chamber 103 and the eluate containing the nucleic acids in the extraction chamber 103 is discharged into the amplification chamber 106.

The extraction cavity 103 is a cavity for extracting nucleic acid, and is communicated with the sample adding cavity 102 through a sample adding cavity liquid micro-channel 1022, and due to the existence of an extraction cavity exhaust channel, a sample solution is extruded by the sample adding sealing plug 200 to flow downwards into the extraction cavity 103 after the user closes the cover during sample adding; the extraction reagent enters the extraction cavity 103 through the first flow passage 1044 under the control of a motor on the detection instrument, and the steps of extracting nucleic acid such as ultrasonic, mixing and heating are realized by matching with an ultrasonic, magnetic attraction and heating module on the instrument. In the extraction process, the extraction cavity 103 is communicated with the waste liquid cavity 105, a flow channel membrane valve waste liquid cavity valve of the extraction cavity 103 and an amplification cavity valve are pressed by an instrument motor to ensure that liquid does not flow downwards, and the motor of the waste liquid cavity valve is loosened when waste liquid is discharged, so that the waste liquid flows downwards into the waste liquid cavity 105 due to the existence of a waste liquid cavity exhaust channel. After the nucleic acid extraction is completed, the eluent enters the extraction chamber 103 through the second flow channel 1045, and after the elution process is completed, the motor of the amplification chamber valve is released, and the eluent containing nucleic acid breaks through the membrane valve formed by the amplification chamber branch flow channel cutting part 1065 and flows into the amplification chamber 106.

Wherein activating the ultrasonic instrument may comprise: after the ultrasonic instrument is matched with the extraction cavity 103 through the ultrasonic hole 302, the ultrasonic instrument is started again.

Further, the fitting of the magnetic attraction instrument with the extraction chamber 103 and the discharge of the waste liquid in the extraction chamber 103 into the waste liquid chamber 105 include:

matching a magnetic attraction instrument with the extraction cavity 103;

the amplification chamber valve is controlled to be kept closed, the waste liquid chamber valve is controlled to be opened, and the waste liquid in the extraction chamber 103 is discharged into the waste liquid chamber 105.

In addition, the coupling of the magnetic attraction instrument with the extraction chamber 103 and the discharge of the nucleic acid-containing eluent in the extraction chamber 103 into the amplification chamber 106 includes:

matching a magnetic attraction instrument with the extraction cavity 103;

the waste liquid cavity valve is controlled to be kept closed, the amplification cavity valve is controlled to be opened, and the eluent containing the nucleic acid in the extraction cavity 103 is discharged into the amplification cavity 106.

Specifically, the instrument motor pushes the piston cap 400, so that the pre-packaged vesicle extraction reagent (lysis solution, proteinase K and magnetic bead mixed solution) in the first storage cavity 1042 is extruded along the first flow channel 1044 into the extraction cavity 103, and is in ultrasonic fit with the ultrasonic instrument to perform the ultrasonic lysis process. Then, the magnetic beads are matched with the instrument magnetic attraction module, the nucleic acid adsorbed by the magnetic beads stays on the wall of the extraction cavity 103, the motor pressing the waste liquid cavity micro-channel cutting part 1056 is removed, the waste liquid enters the waste liquid cavity 105 along the waste liquid cavity micro-channel 1055, and the membrane valve of the waste liquid cavity micro-channel cutting part 1056 is sealed again; then the instrument motor pushes the piston cover 400, so that the pre-packaged vesicle washing liquid reagent in the first storage cavity 1042 is extruded to enter the extraction cavity 103 along the first flow channel 1044, the instrument magnetic suction module is removed, and the magnetic beads adsorbed with nucleic acid are washed by ultrasonic mixing; restarting the instrument magnetic suction module, removing the motor pressing the waste liquid cavity micro-channel cutting part 1056, enabling washing waste liquid to enter the waste liquid cavity 105 along the waste liquid cavity micro-channel 1055, and resealing the membrane valve of the waste liquid cavity micro-channel cutting part 1056; the instrument motor pushes the piston cover 400 to enable the pre-packaged vesicle eluting liquid reagent in the second storage cavity 1043 to be extruded along the second flow channel 1045 to enter the extraction cavity 103, ultrasonic mixing is carried out, the instrument magnetic suction module is restarted, the motor pressing the amplification cavity micro-channel cutting part 1062 is removed, the eluting solution containing nucleic acid enters the amplification cavity main flow channel 10611, the membrane valve of the amplification cavity sub-channel cutting part 1065 is broken, and the eluting solution is injected into the amplification cavity 106 from the bottom along the amplification cavity sub-channel 10612.

Wherein, after injecting the resulting nucleic acid-containing eluate into the plurality of amplification chambers 106, and before starting the fluorescent PCR module, the nucleic acid extraction fluorescent PCR detection method further comprises:

the heating instrument is started to heat the plurality of wax valve cavities 107, so that the wax in the plurality of wax valve cavities 107 is melted and flows downwards into the plurality of amplification cavity branch channels 10611 and the plurality of amplification cavity exhaust micro channels 1063 correspondingly, so as to seal the plurality of amplification cavities 106.

Specifically, the instrument heating module located at the position of the wax valve cavity 107 is started to melt the solid paraffin in the wax valve cavity 107, and fills the solid paraffin downwards along the direction of the amplification cavity exhaust micro-channel 1063 and the amplification cavity branch channels 10612 and 506, so as to seal the amplification cavity exhaust hole 1064, the amplification cavity exhaust micro-channel 1063 and the amplification cavity branch channels 10612, and complete the complete sealing of the amplification cavity 106.

Optionally, step S4 includes:

and starting the instrument fluorescent PCR module, and completing the PCR process of nucleic acid amplification detection through thermal cycling, lighting and detection.

The above description is a chip bottom shell 100, a microfluidic chip 1000 and a nucleic acid extraction fluorescence PCR detection system, the flow channel of the chip bottom shell 100 of the microfluidic chip 1000 and the nucleic acid extraction fluorescence PCR detection system is simple to arrange, a control valve with a complex structure is not required to be arranged, the structure is simple and reasonable, the requirements on design and material use are not high, and the chip bottom shell is an integrated part, and the manufacturing cost is low. In addition, the chip bottom shell provided by the utility model is provided with a plurality of amplification chambers, liquid reagents for extracting nucleic acid by a magnetic bead method and freeze-drying reagents for amplifying nucleic acid can be pre-buried, one consumable material is used, one sample is added at a time, the operation time of personnel is less than one minute, and the fluorescent PCR detection of a plurality of targets can be realized by matching with an instrument to finish the nucleic acid extraction and amplification flow of a sample in and a sample out, so that the detection efficiency is high.

The preferred embodiments of the present utility model have been described in detail above with reference to the accompanying drawings, but the present utility model is not limited to the specific details of the above embodiments, and various simple modifications can be made to the technical solution of the present utility model within the scope of the technical concept of the present utility model, and all the simple modifications belong to the protection scope of the present utility model.

In addition, the specific features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various possible combinations are not described further.

Moreover, any combination of the various embodiments of the utility model can be made without departing from the spirit of the utility model, which should also be considered as disclosed herein.

Claims (10)

1. A chip bottom case of a microfluidic chip, characterized in that the chip bottom case (100) comprises:

a substrate (101);

an extraction chamber (103) provided on the substrate (101); and

detection reagent storage module (104) is provided on base plate (101), detection reagent storage module (104) include a plurality of can with piston lid (400) cooperation and with draw the columnar storage chamber of chamber (103) intercommunication, a plurality of the axis in columnar storage chamber is all followed the face transverse arrangement of base plate (101), a plurality of columnar storage chamber is followed the face longitudinal side by side of base plate (101) arranges, a plurality of piston lid cooperation opening (1041) in columnar storage chamber sets up on base plate thickness face (1013) of base plate (101), base plate thickness face (1013) perpendicular to the face of base plate (101).

2. The chip bottom shell of the microfluidic chip according to claim 1, wherein the chip bottom shell (100) is an integrated part, the chip bottom shell (100) further comprises a sample adding cavity (102), a waste liquid cavity (105) and a plurality of amplification cavities (106) which are respectively communicated with the extracting cavity (103), the substrate (101) comprises a first plate surface and a second plate surface which are oppositely arranged along the thickness direction of the substrate, and the sample adding cavity (102), the extracting cavity (103), the detection reagent storage module (104) and the waste liquid cavity (105) are all arranged on the first plate surface (1011) and are used for communicating a micro-channel between the cavities and a plurality of amplification cavities (106) which are arranged on the second plate surface (1012).

3. The chip bottom case of the microfluidic chip according to claim 2, wherein the sample loading chamber (102) and the extraction chamber (103) are arranged side by side with the detection reagent storage module (104) along the plate surface of the substrate (101) in the lateral direction.

4. The chip bottom case of the microfluidic chip according to claim 2, wherein the sample addition chamber (102), the extraction chamber (103) and the waste liquid chamber (105) are all open-top and the bottom wall of the chamber is disposed on the first plate surface (1011), the peripheral walls of the sample addition chamber (102), the extraction chamber (103) and the waste liquid chamber (105) are all formed on the first plate surface (1011) in a protruding manner, and a plurality of amplification chambers (106) are formed on the second plate surface (1012) in an open-top and concave manner.

5. The chip bottom case of a microfluidic chip according to claim 4, wherein the extraction chamber (103) is arranged adjacent to the detection reagent storage module (104) and shares part of the chamber wall.

6. The microfluidic chip of claim 1, wherein a plurality of the columnar storage cavities comprise:

the first storage cavities (1042) are communicated with the extraction cavity (103) through first flow channels (1044), the first flow channels (1044) comprise first main flow channels and a plurality of first branch flow channels, first ends of the first branch flow channels are respectively communicated with the first storage cavities (1042) one by one, and the first main flow channels are communicated between the extraction cavity (103) and second ends of the first branch flow channels; and

a second storage chamber (1043) communicates with the extraction chamber (103) through a second flow passage (1045).

7. The microfluidic chip bottom case according to claim 6, wherein a plurality of the first branch flow channels are provided with first branch flow channel cutting portions (1046) for cutting off flow channels, and the second flow channels (1045) are provided with second flow channel cutting portions (1047) for cutting off flow channels.

8. A microfluidic chip, characterized in that the microfluidic chip (1000) comprises a chip bottom shell (100) of the microfluidic chip according to any one of claims 2 to 7.

9. The microfluidic chip according to claim 8, wherein the detection reagent storage module (104) is filled with a vesicle extraction reagent, a vesicle wash liquid reagent, and a vesicle elution liquid reagent, the amplification chamber (106) is filled with a nucleic acid amplification freeze-dried detection reagent pellet, and the microfluidic chip (1000) further comprises:

a first coating film welded on one side of the chip bottom shell (100) and used for sealing the waste liquid cavity (105) and the extraction cavity (103);

a second coating film welded on the other side of the chip bottom shell (100) and used for sealing the micro-flow channel of the second plate surface (1012) and a plurality of amplification cavities (106);

a loading plug (200) for closing a loading port of the loading chamber (102) and capable of a piston movement in the loading chamber (102);

the upper cover (300) is arranged on the chip bottom shell (100) in a covering manner and is provided with a sample adding hole (301) and an ultrasonic hole (302), the sample adding hole (301) is aligned with the sample adding cavity (102), and the ultrasonic hole (302) is aligned with the extraction cavity (103); and

a plurality of piston caps (400) are sealed in a one-to-one fit manner in a plurality of the columnar storage chambers.

10. The microfluidic chip according to claim 9, wherein one end of the piston cap (400) is a plunger end, and the other end is provided with a piston rod connection part capable of being connected with a piston rod.