Centrifugal micro-fluidic chip and centrifugal nucleic acid detection system

Abstract

The utility model provides a centrifugal micro-fluidic chip and a centrifugal nucleic acid detection system. The centrifugal micro-fluidic chip comprises: a chip body; the following structures are arranged on the chip body: a liquid injection cavity; the detection modules are arranged at the far end of the liquid injection cavity, each detection module comprises a quantitative cavity, an amplification cavity and a detection cavity which are sequentially arranged from the near end to the far end and communicate with one another, and the near end of each quantitative cavity communicates with the far end of the liquid injection cavity; and a constant voltage module. According to the centrifugal micro-fluidic chip provided by the invention, by arranging the constant-pressure module, the first waste liquid cavity and the constant-pressure flow channel of the constant-pressure module can be used for collecting waste liquid, and meanwhile, when centrifugal detection is carried out, the liquid can smoothly flow from the near end to the far end under the condition that the centrifugal micro-fluidic chip is completely sealed; and the liquid to be detected is prevented from being polluted due to the vent holes and the like in the detection process, so that the detection result is prevented from being influenced.

Description

Technical Field

[0001] The present application relates to the field of nucleic acid detection technology, and in particular to a centrifugal microfluidic chip and a centrifugal nucleic acid detection system. Background Art

[0002] Nucleic acid molecule detection technology, especially detection methods based on polymerase chain reaction (PCR) technology. However, PCR detection technology has relatively high requirements for equipment and cannot meet the needs of instant detection in simple or non-professional scenarios. Isothermal amplification detection technology does not require precise temperature control, has low requirements for equipment, has better sensitivity and specificity than antigen detection, and its detection accuracy is close to PCR detection. Therefore, it is the most commonly used technology in current instant detection products.

[0003] At present, in isothermal amplification detection technology, etc., detection through centrifugal microfluidic chips is already an existing solution. However, as shown in the Chinese utility model application "A centrifugal microfluidic chip integrating nucleic acid extraction and fluorescence amplification" (application number: 202021071424.4), it is necessary to set up vents to ensure the pressure balance in the centrifugal microfluidic chip during use. However, the setting of vents makes the detection environment easily contaminated and affects the accuracy of the detection. Utility Model Content

[0004] In order to solve the problems in the prior art, the present application provides a centrifugal microfluidic chip and a centrifugal nucleic acid detection system. The centrifugal microfluidic chip does not need to be provided with vents, that is, the centrifugal microfluidic chip is in a sealed state during detection, thereby solving the above technical problems. The technical solution of the present application is as follows:

[0005] A centrifugal microfluidic chip, comprising:

[0006] A chip body; and the following structure arranged on the chip body:

[0007] Liquid injection chamber;

[0008] Two or more detection modules, wherein the detection modules are arranged at the distal end of the injection cavity, and the detection modules include a quantitative cavity, an amplification cavity, and a detection cavity which are arranged and connected in sequence from the proximal end to the distal end, and the proximal end of the quantitative cavity is connected to the distal end of the injection cavity; and,

[0009] Constant pressure module;

[0010] Wherein, the constant voltage module comprises:

[0011] More than two first waste liquid chambers, wherein the more than two first waste liquid chambers are located at the far end of the chip body;

[0012] A constant pressure flow channel, the constant pressure flow channel being connected to each of the first waste liquid chambers;

[0013] The proximal end of the injection cavity and the proximal end of the cavity body on the proximal side of the detection cavity are communicated with the constant pressure module.

[0014] Furthermore, at least one opening is provided at the proximal end of the first waste liquid chamber; more than two constant pressure flow channels are connected to each of the first waste liquid chambers through the openings of the first waste liquid chamber to form a closed and surrounding constant pressure module; the injection chamber and the detection module are arranged on the inner side of the constant pressure module.

[0015] Furthermore, the distal end of the quantitative chamber is connected to the proximal end of the amplification chamber through a first siphon flow channel; wherein the proximal end of the first siphon flow channel is located on the proximal side of the proximal end of the quantitative chamber.

[0016] Furthermore, it also includes the following structures arranged on the chip body: a distribution chamber, the distribution chamber is located between the injection chamber and the detection module; a second waste liquid chamber, the second waste liquid chamber is located at the distal end of the distribution chamber; wherein the inlet of the distribution chamber is connected with the injection chamber; the quantitative chamber and the second waste liquid chamber in each of the detection modules are connected with the distribution chamber, and the inlets of the second waste liquid chamber and the distribution chamber are located on different sides of the quantitative chamber of each of the detection modules, so that the liquid flowing into the distribution chamber from the injection chamber first flows into the quantitative chamber in each of the detection modules and then flows into the second waste liquid chamber.

[0017] Furthermore, the centrifugal microfluidic chip further comprises: a connecting chamber, which is arranged between the amplification chamber and the detection chamber, and the connecting chamber is connected to the amplification chamber and the detection chamber respectively through a flow channel.

[0018] Furthermore, the distal end of the amplification chamber is connected to the proximal end of the connection chamber through a second siphon channel; wherein the proximal end of the second siphon channel is located on the proximal side of the proximal end of the amplification chamber.

[0019] Furthermore, the distal end of the connecting cavity is connected to the detection cavity through more than two flow channels.

[0020] Furthermore, the proximal end of the distribution chamber, the proximal end of the amplification chamber and / or the proximal end of the connection chamber are connected to the constant pressure module through a flow channel.

[0021] Furthermore, the centrifugal microfluidic chip is fan-shaped.

[0022] The present application also provides a centrifugal nucleic acid detection system, comprising the centrifugal microfluidic chip described in any of the above items.

[0023] The above-mentioned centrifugal microfluidic chip provided in the present application, by setting a constant pressure module, its first waste liquid chamber and constant pressure flow channel can be used to collect waste liquid. At the same time, when performing centrifugal detection, it can ensure that the liquid flows smoothly from the proximal end to the distal end when the centrifugal microfluidic chip is completely sealed, and prevent the liquid to be tested from being contaminated during the detection process due to the setting of vents, etc., thereby affecting the detection results.

[0024] The above description is only an overview of the technical solution of the present application. In order to make the technical means of the present application clearer and to enable those skilled in the art to implement it according to the contents of the specification, and to make the above and other purposes, features and advantages of the present application more obvious and easy to understand, the specific implementation methods of the present application are exemplified below. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 : A schematic diagram of the structure of a centrifugal microfluidic chip in one embodiment.

[0026] Description of reference numerals:

[0027] 10. Chip body;

[0028] 20. Liquid injection chamber;

[0029] 31. Quantification chamber; 32. Amplification chamber; 33. Detection chamber; 34. Connection chamber;

[0030] 41. a first waste liquid chamber; 42. a constant pressure flow channel;

[0031] 50. The first siphon flow channel;

[0032] 60. Distribution chamber;

[0033] 70. Second waste liquid chamber;

[0034] 80. Second siphon channel. DETAILED DESCRIPTION

[0035] The following embodiments of the present application are only used to illustrate the specific embodiments of the present application, and these embodiments cannot be understood as limiting the present application. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present application are deemed to be equivalent replacement methods and fall within the scope of protection of the present application.

[0036] Those skilled in the art should understand that, in the disclosure of the present application, the terms "first", "second", "third", "fourth", "fifth", etc. are only used to distinguish different structures, but do not limit the number, connection relationship, etc. of specific structures; in addition, the directions or positional relationships indicated by "longitudinal", "transverse", "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc. are based on the directions or positional relationships shown in the accompanying drawings, which are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation. Therefore, the above terms cannot be understood as limitations on the present application.

[0037] In this application, "proximal end" refers to the end of the centrifugal microfluidic chip that is close to the rotating shaft when it is set on the centrifugal nucleic acid detection device. "Distal end" refers to the end of the centrifugal microfluidic chip that is far away from the rotating shaft when it is set on the centrifugal nucleic acid detection device. During centrifugal detection, the liquid in the centrifugal microfluidic chip gradually moves from the proximal chamber to the distal chamber.

[0038] This embodiment provides a centrifugal microfluidic chip, such as Figure 1 As shown, including:

[0039] A chip body 10; and the following structures arranged on the chip body:

[0040] Liquid injection chamber 20;

[0041] Two or more detection modules, wherein the detection modules are arranged at the distal end of the injection chamber 20, and the detection modules include a quantitative chamber 31, an amplification chamber 32, and a detection chamber 33 which are arranged and connected in sequence from the proximal end to the distal end, and the proximal end of the quantitative chamber 31 is connected to the distal end of the injection chamber 20; and,

[0042] Constant pressure module;

[0043] Wherein, the constant voltage module comprises:

[0044] More than two first waste liquid chambers 41, wherein the more than two first waste liquid chambers 41 are located at the far end of the chip body 10;

[0045] A constant pressure channel 42, wherein the constant pressure channel 42 is connected to each of the first waste liquid chambers 41;

[0046] The proximal end of the injection cavity 20 and the proximal end of the cavity on the proximal side of the detection cavity 33 are both connected to the constant pressure module.

[0047] The chip body 10 may include a substrate and a cover layer covering the substrate, wherein each cavity and each flow channel are formed on the substrate, and the cover layer covers the substrate to seal each cavity and each flow channel on the substrate. Specifically, the liquid to be detected may be added to the injection cavity 20, the air-dried reagent / lyophilized reagent for amplification may be added to the amplification cavity 32, the probe reagent may be added to the detection cavity 33, and then the substrate may be covered with the cover layer for sealing, and then detected by the machine (centrifugal nucleic acid detection device).

[0048] Regarding the cavity on the proximal side of the detection cavity, those skilled in the art know that when the amplification cavity 32 is directly connected to the detection cavity 33 through a flow channel, the cavity on the proximal side of the detection cavity is the amplification cavity 32; when, as described below, a connecting cavity 34 is further provided between the amplification cavity 32 and the detection cavity 33, and the connecting cavity 34 is respectively connected to the amplification cavity 32 and the detection cavity 33 through a flow channel, that is, when the amplification cavity 32 is connected to the detection cavity 33 through the connecting cavity 34, the cavity on the proximal side of the detection cavity is the connecting cavity 34.

[0049] The present application has no specific restrictions on the shape of the centrifugal microfluidic chip, but in order to facilitate the simultaneous arrangement of multiple groups of centrifugal microfluidic chips in the centrifugal nucleic acid detection device for centrifugal detection, the centrifugal microfluidic chip is preferably fan-shaped. When the centrifugal microfluidic chip is fan-shaped, multiple centrifugal microfluidic chips can be arranged around the rotating shaft of the centrifugal nucleic acid detection device, so that the centrifugal nucleic acid detection device performs the same treatment on multiple centrifugal microfluidic chips, so as to perform the same detection on multiple centrifugal microfluidic chips at the same time.

[0050] This embodiment provides a new centrifugal microfluidic chip. When the centrifugal microfluidic chip after adding the liquid to be detected and the detection reagent and sealing is set on a centrifugal nucleic acid detection device, the detection can be carried out by the following steps:

[0051] Step 1: Use centrifugal force to transfer the liquid to be tested from the injection chamber 20 to the quantitative chamber 31. The quantitative chamber 31 can quantitatively separate the liquid in the injection chamber 20. The volume of the quantitative chamber 31 determines the upper limit of the subsequent liquid transfer volume.

[0052] Step 2: Using centrifugal force to transfer the liquid to be detected from the quantitative chamber 31 to the amplification chamber 32, and then stop the centrifugation to perform amplification;

[0053] Step 3: After a preset time, restart the centrifugation to transfer the liquid in the amplification chamber 32 to the detection chamber 33, and use an instrument to detect the fluorescence or visible light in the detection chamber 33 to obtain the nucleic acid detection result.

[0054] Among them, in the centrifugal microfluidic chip provided by the present application, the first waste liquid chamber 41 and the constant pressure flow channel 42 in the constant pressure module can not only be used to collect waste liquid, but also, since in the process of liquid transfer from the proximal chamber to the distal chamber, the liquid will push the gas on its distal side to move toward the distal end. At this time, since the proximal end of the injection chamber 20 and the proximal end of the cavity on the proximal side of the detection chamber 33 are connected to the constant pressure module, the gas at the distal end of the liquid can flow back to the proximal side of the liquid through the first waste liquid chamber 41 and / or the constant pressure flow channel 42 of the constant pressure module, thereby ensuring that the liquid flows smoothly from the proximal end to the distal end when the centrifugal microfluidic chip is completely sealed, and preventing the liquid to be tested from being contaminated due to the setting of vents during the detection process, thereby affecting the detection results.

[0055] In one embodiment, Figure 1 As shown, at least one opening is provided at the proximal end of the first waste liquid chamber 41; more than two constant pressure flow channels 42 are connected to each of the first waste liquid chambers 41 through the openings of the first waste liquid chamber 41 to form a closed and surrounding constant pressure module; the injection chamber 20 and the detection module are arranged on the inner side of the constant pressure module.

[0056] Specifically, in this embodiment, when the centrifugal microfluidic chip is fan-shaped, Figure 1 As shown, the first waste liquid chamber 41 and the constant pressure channel 42 are in a closed and surrounding shape arranged at the fan-shaped edge of the centrifugal microfluidic chip, and other chambers are arranged inside it. Therefore, when multiple detection modules are set, the chambers of the detection modules can be conveniently connected with the first waste liquid chamber 41 and / or the constant pressure channel 42 of the constant pressure module, thereby facilitating the collection of waste liquid and balancing the internal pressure of the centrifugal microfluidic chip.

[0057] In one embodiment, Figure 1 As shown, the distal end of the quantitative chamber 31 is connected to the proximal end of the amplification chamber 32 through the first siphon channel 50 ; wherein the proximal end of the first siphon channel 50 is located on the proximal side of the proximal end of the quantitative chamber 31 .

[0058] This embodiment specifically provides a communication method between the distal end of the quantitative chamber 31 and the proximal end of the amplification chamber 32, that is, communication through the first siphon channel 50. When the proximal end of the first siphon channel 50 is located on the proximal side of the proximal end of the quantitative chamber 31, during the centrifugation process in the above step 1, it can be ensured that the liquid in the quantitative chamber 31 will not easily flow to the chamber at its distal end. After stopping the centrifugation, in the above step 2, the first siphon channel 50 is used to attract the liquid to cross the proximal side of the first siphon channel 50, and then the centrifugation is started, so as to transfer the liquid to be detected from the quantitative chamber 31 to the amplification chamber 32.

[0059] In one embodiment, Figure 1As shown, the centrifugal microfluidic chip also includes the following structure arranged on the chip body 10:

[0060] A distribution chamber 60, wherein the distribution chamber 60 is located between the injection chamber 20 and the detection module;

[0061] A second waste liquid chamber 70, wherein the second waste liquid chamber 70 is located at a distal end of the distribution chamber 60;

[0062] Among them, the entrance of the distribution chamber 60 is connected with the injection chamber 20; the quantitative chamber 31 and the second waste liquid chamber 70 in each of the detection modules are connected with the distribution chamber 60, and the entrances of the second waste liquid chamber 70 and the distribution chamber 60 are located on different sides of the quantitative chamber of each of the detection modules so that the liquid flowing into the distribution chamber from the injection chamber first flows into the quantitative chamber in each of the detection modules and then flows into the second waste liquid chamber.

[0063] Then, in the above step 1, when the liquid is moved from the injection chamber 20 to the distal chamber thereof by centrifugation, the liquid first enters the distribution chamber 60 through the inlet of the distribution chamber 60, and then flows into the quantitative chamber 31 in each of the detection modules. After the quantitative chamber 31 is filled with liquid, the excess liquid will flow into the second waste liquid chamber 70. Thus, the quantification of the liquid to be detected and the collection of the excess liquid (waste liquid) are achieved.

[0064] In one embodiment, Figure 1 As shown, the centrifugal microfluidic chip further includes: a connecting chamber 34, which is disposed between the amplification chamber 32 and the detection chamber 33, and the connecting chamber 34 is connected to the amplification chamber 32 and the detection chamber 33 through a flow channel.

[0065] This embodiment provides a centrifugal microfluidic chip in which a connecting chamber 34 is provided between the amplification chamber 32 and the detection chamber 33 .

[0066] In one embodiment, Figure 1 As shown, the distal end of the amplification chamber 32 is connected to the proximal end of the connection chamber 34 through the second siphon channel 80 ; wherein the proximal end of the second siphon channel 80 is located on the proximal side of the proximal end of the amplification chamber 32 .

[0067] This embodiment specifically provides a communication method between the distal end of the amplification chamber 32 and the proximal end of the connecting chamber 34, that is, through the second siphon channel 80. In the above step 2, when the proximal end of the second siphon channel 80 is located on the proximal side of the proximal end of the amplification chamber 32, during the centrifugation process, it can be ensured that the liquid in the amplification chamber 32 will not easily flow to the chamber at its distal end (connecting chamber 34). After stopping the centrifugation, the second siphon channel 80 is used to attract the liquid to cross the proximal side of the second siphon channel 80, and then the centrifugation is started, so as to transfer the liquid from the amplification chamber 32 to the connecting chamber 34.

[0068] In one embodiment, Figure 1 As shown, the distal end of the connecting cavity 34 is connected to the detection cavity 33 through more than two flow channels.

[0069] When the distal end of the connecting cavity 34 is connected to the detection cavity 33 through only one flow channel, it is difficult for the liquid to enter the detection cavity 33 from the connecting cavity 34 due to the gas in the detection cavity 33. When the distal end of the connecting cavity 34 is connected to the detection cavity 33 through two or more flow channels, there is a difference in the liquid pressures of the two or more flow channels during the centrifugation process, so that the liquid can easily flow into the detection cavity 33 from some flow channels (such as one flow channel), and the gas in the detection cavity 33 can flow out of the detection cavity 33 from other flow channels (such as another flow channel), thereby facilitating the transfer of the liquid from the connecting cavity to the detection cavity 33.

[0070] In one embodiment, Figure 1 As shown, the proximal end of the distribution chamber, the proximal end of the amplification chamber and / or the proximal end of the connection chamber are connected to the constant pressure module through a flow channel.

[0071] Through the configuration of this embodiment, it is possible to more conveniently ensure the pressure balance in the centrifugal microfluidic chip, thereby facilitating the transfer of liquid from the proximal chamber to the distal chamber.

[0072] Those skilled in the art know that a centrifugal detection system can be formed by the centrifugal microfluidic chip provided in the present application and the corresponding centrifugal nucleic acid detection equipment, that is, the present application can simultaneously provide a centrifugal nucleic acid detection system, which can include any of the above-mentioned centrifugal microfluidic chips.

[0073] Although the embodiments of the present application are described above, the present application is not limited to the above specific embodiments and application fields, and the above specific embodiments are merely illustrative and instructive, rather than restrictive. A person of ordinary skill in the art can make many forms under the guidance of this specification and without departing from the scope of protection of the claims of the present application, all of which belong to the scope of protection claimed in the present application.

Claims

1. A centrifugal microfluidic chip, characterized in that: include: Chip body; And, the following structure is arranged on the chip body: Liquid injection chamber; Two or more detection modules, wherein the detection modules are arranged at the distal end of the injection cavity, and the detection modules include a quantitative cavity, an amplification cavity, and a detection cavity which are arranged and connected in sequence from the proximal end to the distal end, and the proximal end of the quantitative cavity is connected to the distal end of the injection cavity; and, Constant pressure module; Wherein, the constant voltage module comprises: More than two first waste liquid chambers, wherein the more than two first waste liquid chambers are located at the far end of the chip body; A constant pressure flow channel, the constant pressure flow channel being connected to each of the first waste liquid chambers; The proximal end of the injection cavity and the proximal end of the cavity body on the proximal side of the detection cavity are communicated with the constant pressure module.

2. The centrifugal microfluidic chip according to claim 1, characterized in that: At least one opening is provided at the proximal end of the first waste liquid chamber; Two or more constant pressure flow channels are connected to each of the first waste liquid chambers through the opening of the first waste liquid chamber to form the constant pressure module in a closed and surrounding shape; The liquid injection cavity and the detection module are arranged on the inner side of the constant pressure module.

3. The centrifugal microfluidic chip according to claim 1, characterized in that: The distal end of the quantitative chamber is connected to the proximal end of the amplification chamber through a first siphon flow channel; Wherein, the most proximal end of the first siphon flow channel is located on the proximal side of the most proximal end of the quantitative cavity.

4. The centrifugal microfluidic chip according to claim 1, characterized in that: It also includes the following structure arranged on the chip body: A distribution chamber, the distribution chamber is located between the injection chamber and the detection module; a second waste liquid chamber, the second waste liquid chamber being located at a distal end of the distribution chamber; Wherein, the inlet of the distribution chamber is communicated with the injection chamber; The quantitative chamber and the second waste liquid chamber in each of the detection modules are connected to the distribution chamber, and the entrances of the second waste liquid chamber and the distribution chamber are located on different sides of the quantitative chamber of each of the detection modules, so that the liquid flowing into the distribution chamber from the injection chamber first flows into the quantitative chamber in each of the detection modules and then flows into the second waste liquid chamber.

5. The centrifugal microfluidic chip according to any one of claims 1 to 4, characterized in that: The centrifugal microfluidic chip also includes: A connecting chamber is provided between the amplification chamber and the detection chamber, and the connecting chamber is connected with the amplification chamber and the detection chamber respectively through a flow channel.

6. The centrifugal microfluidic chip according to claim 5, characterized in that: The distal end of the amplification chamber is connected to the proximal end of the connection chamber through a second siphon flow channel; Wherein, the most proximal end of the second siphon flow channel is located on the proximal side of the most proximal end of the amplification chamber.

7. The centrifugal microfluidic chip according to claim 5, characterized in that: The distal end of the connecting cavity is communicated with the detection cavity through more than two flow channels.

8. The centrifugal microfluidic chip according to claim 5, characterized in that: The proximal end of the amplification chamber and / or the proximal end of the connection chamber is communicated with the constant pressure module through a flow channel.

9. The centrifugal microfluidic chip according to claim 4, characterized in that: The proximal end of the distribution chamber is communicated with the constant pressure module through a flow channel.

10. The centrifugal microfluidic chip according to claim 1, characterized in that: The centrifugal microfluidic chip is fan-shaped.

11. A centrifugal nucleic acid detection system, characterized in that: The invention comprises the centrifugal microfluidic chip according to any one of claims 1 to 10.

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