Microfluidic luminescent detection reagent disc and luminescent detection method

Abstract

The application relates to the technical field of sample detection, in particular to a microfluidic luminescence detection reagent disc and a luminescence detection method. The reagent disc comprises a disc body, a flow channel group and a sample detection cavity are arranged on the disc body, the flow channel group comprises a pre-excitation liquid flow channel, an excitation liquid flow channel, a reconstitution liquid flow channel, a cleaning liquid flow channel, a sample liquid flow channel and a waste liquid flow channel, one or more cavities are arranged in each flow channel of the flow channel group, each cavity and the sample detection cavity are connected through micro flow channels, wherein the pre-excitation liquid flow channel, the excitation liquid flow channel, the reconstitution liquid flow channel, the cleaning liquid flow channel and the sample liquid flow channel are connected in parallel and converge at the inlet side of the sample detection cavity, and the waste liquid flow channel is connected to the outlet side of the sample detection cavity. The application can improve the adaptability of the microfluidic luminescence detection reagent disc to different detection items, improve the adaptability to different sample detection analysis scenes, and reduce the cost of different adaptability.

Description

Technical Field

[0001] This application relates to the field of sample detection technology, and in particular to a microfluidic luminescence detection reagent disk and luminescence detection method. Background Technology

[0002] Microfluidic reagent trays typically refer to reagent trays used in microfluidic detection instruments, and may also take the form of microfluidic chips. In existing microfluidic analysis equipment, the microfluidic reagent tray is driven by centrifugal force. The microfluidic reagent tray is disc-shaped, with a fixing hole in the center of the chip. During operation, this fixing hole is mounted on the analyzer's motor shaft, allowing it to rotate at high speed along with the motor shaft. Under the action of centrifugal force, the sample flows from the inner circle to the outer circle of the tray. Multiple interconnected reaction chambers are also arranged along the circumference of the microfluidic reagent tray. Under the action of centrifugal force, the sample sequentially enters each chamber for reaction and is then used for detection. By controlling the speed and direction of rotation, [the following can be achieved / achieved]. The movement / stopping or oscillating mixing of fluids demonstrates that, compared to liquid mixing in test tubes, the microfluidic reagent tray is more complex in its fluid control process. Therefore, the channel design is generally based on a single channel serving a single detection item. For example, Chinese Patent CN103323605B provides a microfluidic reagent tray for the immunoassay of glycated hemoglobin in serum, which includes multiple interconnected and centrally symmetrically distributed channels. Although it can detect multiple samples simultaneously, each channel completes the detection operation independently. Therefore, the same microfluidic reagent tray can only perform detection of the same type or the same physicochemical properties, and is not suitable for complex detection items.

[0003] In summary, existing microfluidic analysis equipment has the following drawbacks: the flow channel configuration of microfluidic reagent trays is limited and difficult to apply to complex detection projects such as multi-stage luminescence detection; the same microfluidic reagent tray can only perform detection of the same type of project, and cannot meet multiple detection needs at the same time; and it has weak adaptability to different sample detection and analysis scenarios. Summary of the Invention

[0004] In view of the shortcomings of the prior art, the purpose of this application is to provide a microfluidic luminescence detection reagent disk and luminescence detection method to solve the problems that the existing microfluidic reagent disk has a single flow channel setting method, which is difficult to apply to complex detection projects such as multi-process luminescence detection. The same microfluidic reagent disk can only perform detection of the same type of project, which cannot meet multiple detection needs at the same time, and has weak adaptability to different sample detection and analysis scenarios.

[0005] To achieve the above and other related objectives, in a first aspect, this application provides a microfluidic luminescent detection reagent disk, comprising:

[0006] The disc body is provided with a flow channel group and a sample detection chamber. The flow channel group includes a pre-excitation liquid flow channel, an excitation liquid flow channel, a rehydration solution flow channel, a cleaning liquid flow channel, a sample liquid flow channel, and a waste liquid flow channel. Each flow channel in the flow channel group includes one or more chambers. Each chamber and the sample detection chamber are connected by microchannels. The pre-excitation liquid flow channel, the excitation liquid flow channel, the rehydration solution flow channel, the cleaning liquid flow channel, and the sample liquid flow channel are connected in parallel and converge at the inlet side of the sample detection chamber. The waste liquid flow channel is connected to the outlet side of the sample detection chamber.

[0007] Furthermore, the microfluidic luminescence detection reagent disk is divided into multiple zones, and multiple groups of flow channels are provided, with each flow channel group corresponding to one another in each zone.

[0008] Furthermore, the microfluidic luminescent detection reagent disk is divided into a sample detection area and a calibration detection area within the location of each of the flow channel groups. In each flow channel group, in addition to the sample liquid flow channel, each of the other flow channels is provided with two groups, which are respectively located in the sample detection area and the calibration detection area. The sample detection cavity is located in the sample detection area, and the calibration detection area is also provided with a calibration detection cavity.

[0009] Furthermore, the pre-excitation liquid flow channel includes a pre-excitation liquid storage chamber, a pre-excitation liquid metering chamber, and a pre-excitation liquid quality control chamber, wherein the pre-excitation liquid storage chamber, the pre-excitation liquid metering chamber, and the sample detection chamber are connected in series, and the pre-excitation liquid quality control chamber is connected to the pre-excitation liquid metering chamber.

[0010] Furthermore, the excitation fluid channel includes an excitation fluid storage chamber connected to the sample detection chamber.

[0011] Furthermore, the reconstitution solution flow channel includes a reconstitution solution storage chamber, a reconstitution solution quantification chamber, and a reconstitution solution quality control chamber, wherein the reconstitution solution storage chamber, the reconstitution solution quantification chamber, and the sample detection chamber are connected in series, and the reconstitution solution quality control chamber is connected to the reconstitution solution quantification chamber.

[0012] Furthermore, the cleaning fluid flow channel includes a cleaning fluid storage chamber, a cleaning fluid metering chamber, and a cleaning fluid quality control chamber. The cleaning fluid storage chamber, the cleaning fluid metering chamber, and the sample detection chamber are connected in series, and the cleaning fluid quality control chamber is connected to the cleaning fluid metering chamber.

[0013] Furthermore, multiple cleaning fluid storage chambers are arranged in series, and multiple cleaning fluid quality control chambers are arranged in series, with the number of cleaning fluid storage chambers being the same as the number of cleaning fluid quality control chambers.

[0014] Furthermore, the sample liquid flow channel includes a plasma receiving chamber, a first plasma separation chamber, a second plasma separation chamber, and a plasma quality control chamber. The plasma receiving chamber, the first plasma separation chamber, and the second plasma separation chamber are connected in series. The sample detection chamber is connected in series between the first plasma separation chamber and the second plasma separation chamber. The plasma quality control chamber is connected to the first plasma separation chamber.

[0015] Secondly, this application also provides a luminescence detection method, which controls the microfluidic luminescence detection reagent disk as described above to rotate at different speeds to achieve liquid flow control within the flow channel assembly and the sample detection chamber. The luminescence detection method specifically includes the following steps:

[0016] S1, inject the sample liquid to be tested into the sample liquid channel, inject the pre-activation liquid, activation liquid, reconstitution solution and cleaning liquid into the pre-activation liquid channel, the activation liquid channel, the reconstitution solution channel and the cleaning liquid channel respectively, and place the reagent to be reacted into the sample detection chamber. The sample liquid to be tested includes plasma and the reagent to be reacted includes lyophilized reagent.

[0017] S2, the microfluidic luminescent detection reagent disk is mounted on a rotating stage, the rotating stage being used to drive the microfluidic luminescent detection reagent disk to rotate;

[0018] S3, the rotating stage drives the microfluidic luminescent detection reagent disk to rotate with a first rotation speed parameter, so that the reconstitution solution is quantitatively released into the sample detection chamber and mixed and incubated with the reagent to be reacted;

[0019] S4, the rotary stage drives the microfluidic luminescent detection reagent disk to rotate with a second rotation speed parameter, so that the liquid sample to be tested is quantitatively released into the sample detection chamber;

[0020] S5, after the reconstitution solution and the sample liquid to be tested are fully mixed in the sample detection chamber, the rotating stage drives the microfluidic luminescent detection reagent disk to rotate with the third rotation speed parameter, so that the cleaning solution is quantitatively released into the sample detection chamber;

[0021] S6, the rotary table drives the microfluidic luminescent detection reagent disk to rotate with a fourth rotational speed parameter, so that the mixed liquid in the sample detection chamber is discharged into the waste liquid channel;

[0022] S7, the rotating stage drives the microfluidic luminescent detection reagent disk to rotate with the fifth rotation speed parameter, so that the pre-excitation liquid is quantitatively released into the sample detection chamber and mixed with the reagent to be reacted for incubation;

[0023] S8, the rotary stage drives the microfluidic luminescent detection reagent disk to rotate with the sixth rotation speed parameter, so that the excitation liquid is quantitatively released into the sample detection chamber to generate a light signal;

[0024] S9, the optical signal is collected for component analysis.

[0025] The microfluidic luminescence detection reagent tray and luminescence detection method described above, by setting a flow channel group and a sample detection chamber on the microfluidic reagent tray, wherein the flow channel group includes a pre-excitation liquid flow channel, an excitation liquid flow channel, a reconstitution solution flow channel, a cleaning liquid flow channel, a sample liquid flow channel, and a waste liquid flow channel, and the chambers in each flow channel are connected by microchannels, so that the flow control of liquid in different chambers can be completed at different rotation speeds. This allows for the completion of various complex detection items such as luminescence detection by arranging and combining fluid control schemes in multiple channels and chambers, effectively meeting the requirements of different detection steps and detection items, effectively improving the adaptability of the microfluidic luminescence detection device to different detection items, greatly improving the adaptability to differential sample detection and analysis scenarios, and reducing the differential adaptation cost of the microfluidic reagent tray. Attached Figure Description

[0026] Figure 1 This is a schematic diagram of the overall structure of a microfluidic luminescent detection reagent disk, as shown in an exemplary embodiment of this application.

[0027] Figure 2 This is a schematic diagram illustrating the flow channel division within a section of a microfluidic luminescent detection reagent disk, as shown in an exemplary embodiment of this application.

[0028] Figure 3 This is a schematic flowchart illustrating a light emission detection method as an exemplary embodiment of this application.

[0029] Part Number Explanation

[0030] 1-Disc body; 10-Sample detection chamber; 100-Lyophilized reagent sphere; 101-Sample detection area; 102-Calibration detection area; 11-Pre-excitation fluid channel; 111-Pre-excitation fluid storage chamber; 112-Pre-excitation fluid quantitative chamber; 113-Pre-excitation fluid quality control chamber; 12-Excitation fluid channel; 121-Excitation fluid storage chamber; 13-Reconstitution solution channel; 131-Reconstitution solution storage chamber; 132-Reconstitution solution quantitative chamber; 133-Reconstitution solution quality control chamber; 14-Washing fluid channel; 141-Washing fluid storage chamber; 142-Washing fluid quantitative chamber; 143-Washing fluid quality control chamber; 15-Sample liquid channel; 151-Plasma container chamber; 152-Plasma first separation chamber; 153-Plasma second separation chamber; 154-Plasma quality control chamber; 16-Waste liquid channel; 17-Calibration detection chamber. Detailed Implementation

[0031] The following specific embodiments illustrate the implementation of this application. Those skilled in the art can easily understand other advantages and effects of this application from the content disclosed in this specification.

[0032] It should be understood that the structures, proportions, sizes, etc., illustrated in the accompanying drawings of this specification are merely for illustrative purposes to aid those skilled in the art and are not intended to limit the scope of this application. Therefore, they have no substantial technical significance. Any modifications to the structure, changes in proportions, or adjustments to size, without affecting the effectiveness and purpose of this application, should still fall within the scope of the technical content disclosed in this application. Furthermore, the terms such as "upper," "lower," "left," "right," "middle," "first," and "second" used in this specification are merely for clarity and are not intended to limit the scope of this application. Changes or adjustments to their relative relationships, without substantially altering the technical content, should also be considered within the scope of this application.

[0033] In one embodiment, this application exemplarily illustrates a microfluidic luminescent detection reagent disk; please refer to [link to relevant documentation]. Figures 1-2 The microfluidic luminescence detection reagent disk includes a disk body 1. In this embodiment, the disk body 1 is generally disc-shaped. The disk body 1 is provided with a flow channel assembly and a sample detection chamber 10. The flow channel assembly includes a pre-excitation liquid flow channel 11, an excitation liquid flow channel 12, a rehydration solution flow channel 13, a cleaning liquid flow channel 14, a sample liquid flow channel 15, and a waste liquid flow channel 16. Each flow channel in the flow channel assembly includes one or more chambers. The chambers and the sample detection chamber 10 are connected by microfluidic channels. The pre-excitation liquid flow channel 11, the excitation liquid flow channel 12, the rehydration solution flow channel 13, the cleaning liquid flow channel 14, and the sample liquid flow channel 15 are connected by microfluidic channels. Liquid channels 15 are connected in parallel and converge at the inlet side of the sample detection chamber 10. Waste liquid channel 16 is connected to the outlet side of the sample detection chamber 10. It is worth noting that microchannels refer to fluid flow paths with micrometers as the unit of measurement. Depending on the properties of the fluid itself, different microchannel sizes and different rotation speeds will affect the fluid's permeability within the microchannel. Thus, based on different preset rotation speeds, the flow control of fluids in different channels on the same disc 1 can be achieved. Furthermore, according to the preset rotation speed arrangement, the fluid flow actions required for different detection procedures can be executed.

[0034] As can be seen, in the microfluidic luminescence detection reagent disk provided in this application, by setting a flow channel group and a sample detection chamber 10 on the disk body 1, the flow channel group includes a pre-excitation liquid flow channel 11, an excitation liquid flow channel 12, a reconstitution solution flow channel 13, a cleaning liquid flow channel 14, a sample liquid flow channel 15, and a waste liquid flow channel 16. The chambers in each flow channel are connected by microfluidic channels, so that the flow control of liquid in different chambers can be completed at different rotation speeds. This allows for the completion of various complex detection items such as luminescence detection through the arrangement and combination of fluid control schemes in multiple channels and chambers, effectively meeting the needs of different detection steps and detection items, effectively improving the adaptability of the microfluidic luminescence detection device to different detection items, greatly improving the adaptability to differential sample detection and analysis scenarios, and reducing the differential adaptation cost of the disk body 1.

[0035] In this embodiment, please refer to Figure 1 The microfluidic luminescence detection reagent disk is divided into multiple zones, and multiple sets of flow channels are set, with each flow channel set corresponding to one another in each zone. Specifically, the disk body is a disk formed by splicing two semi-circular regions, and each semi-circular region is a zone, on which flow channels are set respectively. Therefore, the flow channels are set in two centrally symmetrical groups along the circumference of the disk body 1, located in the two semi-circular regions respectively. In some other embodiments, the flow channels can also be set in three or more groups on the same disk according to their size and design requirements, thereby dividing the microfluidic luminescence detection reagent disk into three or more zones so as to perform simultaneous detection of multiple groups of samples.

[0036] In this embodiment, the disk 1 is divided into a sample detection area 101 and a calibration detection area 102 within each flow channel group. In each flow channel group, except for the sample liquid flow channel 15, each other flow channel is provided with two groups, one of which is located in the sample detection area 101 and the calibration detection area 102. The sample detection chamber 10 is located in the sample detection area 101, and the calibration detection area 102 is also provided with a calibration detection chamber 17. The calibration detection chamber 17 is used to place the same reagent as in the sample detection area 101, such as the lyophilized reagent ball 100, but no sample liquid enters the calibration detection area 102. It can be understood that the calibration detection area 102 is detected synchronously with the sample detection area 101, and its detection result does not involve the sample liquid. Therefore, its detection result is used as a control group or for calibration.

[0037] In this embodiment, the pre-excitation liquid flow channel 11 includes a pre-excitation liquid storage chamber 111, a pre-excitation liquid metering chamber 112, and a pre-excitation liquid quality control chamber 113. The pre-excitation liquid storage chamber 111 stores the pre-excitation liquid. The pre-excitation liquid storage chamber 111, the pre-excitation liquid metering chamber 112, and the sample detection chamber 10 are connected in series. The pre-excitation liquid quality control chamber 113 is connected to the pre-excitation liquid metering chamber 112. At a specific rotation speed, the pre-excitation liquid enters the pre-excitation liquid metering chamber 112 and the pre-excitation liquid quality control chamber 113 from the pre-excitation liquid storage chamber 111 through the microchannel. The pre-excitation liquid metering chamber 112 is used to quantitatively store the pre-excitation liquid required in one detection process.

[0038] In this embodiment, in some application scenarios, detection can be performed by photosensitive components, such as photosensors, arranged along the rotation path of the pre-excitation liquid quality control cavity 113. As the pre-excitation liquid quality control cavity 113 rotates with the disk body 1, it receives light signal detection at at least one position along its rotation path. Based on the detection results, the liquid state of the pre-excitation liquid in the pre-excitation liquid quality control cavity 113 can be known, and the current flow state of the pre-excitation liquid can be inferred, which plays a guiding role in controlling the rotation speed of the microfluidic luminescent detection reagent disk.

[0039] In some application scenarios, the rotation control process of disk 1 is as follows:

[0040] The disk 1 rotates at a first preset speed, and the pre-excitation liquid enters the pre-excitation liquid metering chamber 112. After the pre-excitation liquid metering chamber 112 is full, it continues to rotate, and the pre-excitation liquid enters the pre-excitation liquid quality control chamber 113.

[0041] The liquid state of the pre-excitation liquid quality control chamber 113 is detected. After the pre-excitation liquid quality control chamber 113 is full, the disk 1 rotates at the second preset speed. The pre-excitation liquid enters the sample detection chamber 10 from the pre-excitation liquid quantitative chamber 112 and mixes with the detection sample for pre-excitation.

[0042] As can be seen, by utilizing the structure of the microfluidic luminescent detection reagent disk shown in this embodiment, the fluid flow state within the pre-excitation liquid channel 11 can be precisely controlled to meet the needs of different detection processes. It is understood that in other channels and chambers, such as the reconstitution solution channel 13 and the cleaning liquid channel 14, the fluid flow state can be precisely controlled through the same action logic. The above control process is merely an exemplary description for the purpose of understanding this solution and is not intended to limit the application of this solution. In other embodiments, other application methods can also be adopted based on the structure of the microfluidic luminescent detection reagent disk shown in this application.

[0043] In this embodiment, the excitation fluid channel 12 includes an excitation fluid storage cavity 121 connected to the sample detection cavity 10. The excitation fluid storage cavity 121 is used to store excitation fluid so as to excite different samples in the sample detection cavity 10 and generate different light emission characteristics. In some application scenarios, the light signals generated after different samples are excited can be collected by photosensitive components such as photosensors to obtain the corresponding detection information.

[0044] In this embodiment, the reconstitution channel 13 includes a reconstitution storage chamber 131, a reconstitution quantitative chamber 132, and a reconstitution quality control chamber 133. The reconstitution storage chamber 131, the reconstitution quantitative chamber 132, and the sample detection chamber 10 are connected in series, and the reconstitution quality control chamber 133 is connected to the reconstitution quantitative chamber 132.

[0045] In this embodiment, the cleaning fluid channel 14 includes a cleaning fluid storage chamber 141, a cleaning fluid metering chamber 142, and a cleaning fluid quality control chamber 143. The cleaning fluid storage chamber 141, the cleaning fluid metering chamber 142, and the sample detection chamber 10 are connected in series. The cleaning fluid quality control chamber 143 is connected to the cleaning fluid metering chamber 142.

[0046] In this embodiment, multiple cleaning fluid storage chambers 141 are arranged in series, for example, three, and multiple cleaning fluid quality control chambers 143 are arranged in series, for example, three. The number of cleaning fluid storage chambers 141 is the same as the number of cleaning fluid quality control chambers 143. Multiple cleaning fluid quality control chambers 143 and cleaning fluid storage chambers 141 can meet the needs of different cleaning fluid volumes and different cleaning fluid release sequences in different testing processes.

[0047] In this embodiment, the sample liquid flow channel 15 includes a plasma receiving chamber 151, a first plasma separation chamber 152, a second plasma separation chamber 153, and a plasma quality control chamber 154. The plasma receiving chamber 151, the first plasma separation chamber 152, and the second plasma separation chamber 153 are connected in series. The sample detection chamber 10 is connected in series between the first plasma separation chamber 152 and the second plasma separation chamber 153. The plasma quality control chamber 154 is connected to the first plasma separation chamber 152.

[0048] The first plasma separation chamber 152 and the second plasma separation chamber 153 are used to centrifuge the plasma during the rotation of the disc 1, so that some components are separated into the second plasma separation chamber 153, thereby meeting the requirements for plasma component separation during the luminescence detection process.

[0049] In the above embodiments, the action of detecting optical signals in each quality control cavity, such as the pre-excitation liquid quality control cavity 113, the reconstitution solution quality control cavity 133, the cleaning liquid quality control cavity 143, and the plasma quality control cavity 154, can all be accomplished by photosensitive components, such as photosensors, arranged toward the quality control cavity. The structure shown in this embodiment is sufficient to provide the flow channel structure basis required for complex detection items, so no further limitations are made.

[0050] Furthermore, in some other embodiments, in order to facilitate the acquisition of fluid flow conditions in each channel, quantitative chambers and quality control chambers can be connected at other locations in each channel. For example, in this embodiment, quantitative chambers and quality control chambers are also connected at the inlet ends of the sample detection chamber 10 and the calibration detection chamber 17, and additional quality control chambers are connected on the sample liquid channel 15 and the waste liquid channel 16.

[0051] In summary, the microfluidic luminescence detection reagent disk provided in this application, by setting a flow channel group and a sample detection chamber 10 on the disk body 1, wherein the flow channel group includes a pre-excitation liquid flow channel 11, an excitation liquid flow channel 12, a reconstitution solution flow channel 13, a cleaning liquid flow channel 14, a sample liquid flow channel 15, and a waste liquid flow channel 16, and the chambers in each flow channel are connected by microchannels, so that the flow control of the liquid in different chambers can be completed at different rotation speeds. This allows for the completion of various complex detection items such as luminescence detection by arranging and combining fluid control schemes in multiple channels and chambers, effectively meeting the requirements of different detection steps and detection items, effectively improving the adaptability of the microfluidic luminescence detection device to different detection items, greatly improving the adaptability to differential sample detection and analysis scenarios, and reducing the differential adaptation cost of the disk body 1.

[0052] In another embodiment, this application also discloses a luminescence detection method, which controls the liquid flow within the flow channel assembly and sample detection chamber by controlling the microfluidic luminescence detection reagent disk in the foregoing embodiments to rotate at different speeds. Please refer to [link to relevant documentation]. Figure 3 The luminescence detection method includes the following steps:

[0053] S1, inject the sample liquid to be tested into the sample liquid channel, inject the pre-activation liquid, activation liquid, reconstitution solution and cleaning liquid into the pre-activation liquid channel, activation liquid channel, reconstitution solution channel and cleaning liquid channel respectively, and place the reagent to be reacted into the sample detection chamber. The sample liquid to be tested includes plasma and the reagent to be reacted includes lyophilized reagent.

[0054] S2, The microfluidic luminescence detection reagent disk is mounted on the rotating stage, and the rotating stage is used to drive the microfluidic luminescence detection reagent disk to rotate;

[0055] S3, the rotating stage drives the microfluidic luminescent detection reagent disk to rotate with the first rotation speed parameter, so that the reconstituted solution is quantitatively released into the sample detection chamber and mixed with the reagent to be reacted for incubation;

[0056] S4, the rotary stage drives the microfluidic luminescent detection reagent disk to rotate with the second rotation speed parameter, so that the liquid sample to be tested is quantitatively released into the sample detection chamber;

[0057] S5. After the reconstitution solution and the sample liquid to be tested are fully mixed in the sample detection chamber, the rotating stage drives the microfluidic luminescent detection reagent disk to rotate at the third rotation speed parameter so that the cleaning solution is quantitatively released into the sample detection chamber.

[0058] S6, the rotary stage drives the microfluidic luminescent detection reagent disk to rotate with the fourth rotation speed parameter, so that the mixed liquid in the sample detection chamber is discharged to the waste liquid channel;

[0059] S7, the rotating stage drives the microfluidic luminescent detection reagent disk to rotate with the fifth rotation speed parameter, so that the pre-excitation liquid is quantitatively released into the sample detection chamber and mixed with the reagent to be reacted for incubation;

[0060] S8, the rotary stage drives the microfluidic luminescent detection reagent disk to rotate with the sixth rotation speed parameter, so that the excitation liquid is quantitatively released into the sample detection chamber to generate a light signal;

[0061] S9 collects optical signals for component analysis.

[0062] Regarding step S2, it should be understood that the rotary table is a device in the art used to mount and fix reagent trays or microfluidic chips and drive them to rotate. Its principle is that a driving component that outputs torque, such as a motor, drives the mounting component to rotate, and then the reagent tray or microfluidic chip is detachably mounted to the mounting component and rotates together. Since there are various types of existing driving components that can output torque, they are not limited here. The mounting component can be, for example, a clamp, magnetic component, chuck, or other structure that can detachably fix the reagent tray, connected to the output end of the driving component. For example, in this embodiment, the rotary table includes a motor and a chuck connected to the output end of the motor. The chuck is provided with a buckle for locking the reagent tray. The motor's switch and speed adjustment can be manually operated or controlled by an automation module. For example, in some embodiments, the driving component in the rotary table can also be connected to an assembly with a control program, such as a PLC motherboard, or a computer with control software installed to achieve automated control of the rotation.

[0063] In this embodiment, before executing step S6, that is, before the rotary table drives the microfluidic luminescent detection reagent disk to rotate at the fifth rotation speed parameter so that the pre-excitation liquid is quantitatively released into the sample detection chamber and mixed with the reagent to be reacted for incubation, the sample detection chamber is cleaned multiple times by repeating steps S5 and S6. For example, based on the structure of the microfluidic luminescent detection reagent disk shown in the previous embodiment, three cleaning liquid storage chambers 141 and three cleaning liquid quality control chambers 143 are provided in each cleaning liquid flow channel 14. By controlling the rotation speed, the cleaning liquid in the three cleaning liquid storage chambers 141 enters the quantitative chamber and the quality control chamber in sequence.

[0064] In this embodiment, one or more channels in the flow channel group are provided with a quantitative chamber and a quality control chamber. In the luminescence detection method, in steps S3-S8, the fluid can be quantitatively released through the quantitative chamber, and the fluid storage in the quantitative chamber can be judged through the quality control chamber.

[0065] It is worth noting that in the above embodiments, the first to sixth speed parameters are not quantitative descriptions of the speed, but rather qualitative classifications. That is, each speed parameter may include multiple speed values ​​used to complete its respective step once or multiple times. For example, in the aforementioned embodiments, during the repeated execution of steps S5 and S6, the cleaning fluid enters the sample detection chamber 10 from the three cleaning fluid storage chambers 141 in three separate steps. For instance, in the first execution of step S5, the cleaning fluid in the first cleaning fluid storage chamber is rotated at speed A1, and then enters the cleaning fluid metering chamber 142 and the first cleaning fluid quality control chamber. Then, the cleaning fluid in the cleaning fluid metering chamber 142 is rotated at speed A2, and then enters the sample detection chamber 10. In the second execution of step S5, the cleaning fluid in the second cleaning fluid storage chamber is rotated at speed B1, and then enters the sample detection chamber 10 from the cleaning fluid metering chamber 142. The cleaning solution enters the cleaning solution metering chamber 142 and the second cleaning solution quality control chamber. Then, it rotates at a speed of B2, and the cleaning solution in the cleaning solution metering chamber 142 enters the sample detection chamber 10. The third step S5 is executed. First, it rotates at a speed of C1, and the cleaning solution in the third cleaning solution storage chamber enters the cleaning solution metering chamber 142 and the third cleaning solution quality control chamber. Then, it rotates at a speed of C2, and the cleaning solution in the cleaning solution metering chamber 142 enters the sample detection chamber 10. During the three executions of step S5, the cleaning solution in the three cleaning solution storage chambers 141, i.e., the first to third cleaning solution storage chambers, is released in sequence, and the three cleaning solution quality control chambers 143, i.e., the first to third cleaning solution quality control chambers, are filled with cleaning solution in sequence. The speeds A1, A2, B1, B2, C1, and C2 together constitute the third speed parameter.

[0066] It is worth noting that, since the microfluidic luminescent detection reagent disk is equipped with microchannels, different microchannel sizes and different rotation speeds will affect the fluid's permeability within the microchannels depending on the fluid's properties. This allows for flow control of the fluid in different channels based on different preset rotation speeds. Furthermore, by arranging the fluids in a preset rotation speed sequence, different fluid flow actions required for different detection processes can be performed. Therefore, in this embodiment or other embodiments, the fluid control method in other channels can be the same as the control method for the cleaning fluid in the above embodiment, and asynchronous flow of liquids in different channels can be achieved by changing the rotation speed of the rotary table.

[0067] In other embodiments, microswitches of various types in the art can be installed in the flow channel to assist in controlling the opening and closing of the flow channel.

[0068] In some implementations, the monitoring of the liquid state inside various channels, such as the quality control chamber, can be achieved not only through manual observation but also through radar, optical sensors, and temperature sensors. The data is then aggregated and processed through information processing ports such as processors, CPUs, and PLC motherboards to guide the rotation speed switching of the rotary table. This integrates the microfluidic luminescence detection reagent disk into an independent detection instrument or device, enabling the instrument or device to fully execute the luminescence detection method of this embodiment according to a pre-programmed control program.

[0069] In this embodiment, the microfluidic luminescence detection reagent disk is also provided with a calibration detection chamber 17. Based on the description of the application scenario in the aforementioned embodiments, it is easy to understand that during steps S1-S9, except that no sample liquid needs to be injected, the same fluid control actions will occur in the flow channels within the calibration detection area 102 as in the sample detection chamber 101. The flow control of the pre-excitation liquid, excitation liquid, reconstitution solution, and cleaning liquid is the same. For details, please refer to... Figure 3 The illustrated flowchart of the light emission detection method shows that, after step S2 and before step S9, the light emission detection method further includes the following steps:

[0070] S31, the rotating stage drives the microfluidic luminescent detection reagent disk to rotate with the first rotation speed parameter, so that the reconstitution solution is quantitatively released into the calibration detection chamber and mixed with the reagent to be reacted for incubation;

[0071] S51, after the reconstitution solution and the sample liquid to be tested are fully mixed in the calibration detection chamber, the rotary stage drives the microfluidic luminescent detection reagent disk to rotate at the third rotation speed parameter so that the cleaning solution is quantitatively released into the calibration detection chamber;

[0072] S61, the rotary stage drives the microfluidic luminescent detection reagent disk to rotate with the fourth rotation speed parameter, so that the mixed liquid in the calibration detection chamber is discharged to the waste liquid channel;

[0073] S71, the rotary stage drives the microfluidic luminescent detection reagent disk to rotate with the fifth rotational speed parameter, so that the pre-excitation liquid is quantitatively released into the calibration detection chamber and mixed with the reagent to be reacted for incubation;

[0074] S81, the rotary stage drives the microfluidic luminescent detection reagent disk to rotate at the sixth rotation speed parameter, so that the excitation liquid is quantitatively released into the calibration detection chamber to generate a light signal.

[0075] Among them, steps S31 and S51-S81 can be executed synchronously or asynchronously with steps S3-S8 between steps S2 and S9. In this embodiment, step S31 can be performed synchronously with step S3, step S51 can be performed synchronously with step S5, step S61 can be performed synchronously with step S6, step S71 can be performed synchronously with step S7, and step S81 can be performed synchronously with step S8.

[0076] It is understood that in this embodiment, the light signals collected in step S9 are at least two sets, which are respectively from the sample detection cavity 10 and the calibration detection cavity 17. The difference between the light signals collected from the two is due to the presence or absence of the liquid in the sample to be tested. Since there is a difference in the presence or absence of the liquid in the sample to be tested, there is also a difference in the collected light signals. Therefore, the light signal collected from the calibration detection cavity 17 in step S9 can be used for calibration or comparative analysis to ensure the accuracy of the detection results in the sample detection cavity 10.

[0077] As described above, the luminescence detection method in this embodiment utilizes the microfluidic luminescence detection reagent disk shown in the previous embodiment. By setting a flow channel group and a sample detection chamber 10 on the disk body 1, the flow channel group includes a pre-excitation liquid flow channel 11, an excitation liquid flow channel 12, a reconstitution solution flow channel 13, a cleaning liquid flow channel 14, a sample liquid flow channel 15, and a waste liquid flow channel 16. The chambers in each flow channel are connected through microchannels, enabling the flow control of liquids in different chambers at different rotation speeds. This allows for the combination of fluid control schemes in multiple channels and chambers to complete various complex detection items such as luminescence detection, effectively meeting the requirements of different detection steps and items. It effectively improves the adaptability of the microfluidic luminescence detection device to different detection items, greatly enhances the adaptability to differential sample detection and analysis scenarios, and reduces the cost of differential adaptation of the disk body 1.

[0078] The above embodiments are merely illustrative of the principles and effects of this application and are not intended to limit this application. Any person skilled in the art can modify or alter the above embodiments without departing from the spirit and scope of this application. Therefore, all equivalent modifications or alterations made by those skilled in the art without departing from the spirit and technical concept disclosed in this application should still be covered by the claims of this application.

Claims

1. A method for detecting luminescence, characterized in that, The detection method controls the flow of liquid within the flow channel group and the sample detection chamber by rotating the microfluidic luminescent detection reagent disk at different speeds. The microfluidic luminescent detection reagent disk includes a disk body, on which a flow channel group and a sample detection chamber are disposed. The flow channel group includes a pre-excitation liquid flow channel, an excitation liquid flow channel, a reconstitution solution flow channel, a cleaning liquid flow channel, a sample liquid flow channel, and a waste liquid flow channel. Each flow channel in the flow channel group includes one or more chambers. Each chamber and the sample detection chamber are connected by a microfluidic channel. The pre-excitation liquid flow channel, the excitation liquid flow channel, the reconstitution solution flow channel, the cleaning liquid flow channel, and the sample liquid flow channel are connected in parallel and converge at the inlet side of the sample detection chamber. The waste liquid flow channel is connected to the outlet side of the sample detection chamber. The microfluidic luminescent detection reagent disk is divided into multiple zones, and multiple groups of flow channels are provided, with each flow channel group corresponding to one another in each zone. The microfluidic luminescent detection reagent disk is divided into a sample detection area and a calibration detection area within the location of each of the flow channel groups. In each flow channel group, except for the sample liquid flow channel, each of the other flow channels is provided with two groups, which are respectively located in the sample detection area and the calibration detection area. The sample detection cavity is located in the sample detection area, and the calibration detection area is also provided with a calibration detection cavity. The detection method includes the following steps: S1, inject the sample liquid to be tested into the sample liquid channel, inject the pre-activation liquid, activation liquid, reconstitution solution and cleaning liquid into the pre-activation liquid channel, the activation liquid channel, the reconstitution solution channel and the cleaning liquid channel respectively, and place the reagent to be reacted into the sample detection chamber. The sample liquid to be tested includes plasma and the reagent to be reacted includes lyophilized reagent. S2, the microfluidic luminescent detection reagent disk is mounted on a rotating stage, the rotating stage being used to drive the microfluidic luminescent detection reagent disk to rotate; S3, the rotating stage drives the microfluidic luminescent detection reagent disk to rotate with a first rotation speed parameter, so that the reconstitution solution is quantitatively released into the sample detection chamber and mixed and incubated with the reagent to be reacted; S4, the rotary stage drives the microfluidic luminescent detection reagent disk to rotate with a second rotation speed parameter, so that the liquid sample to be tested is quantitatively released into the sample detection chamber; S5, after the reconstitution solution and the sample liquid to be tested are fully mixed in the sample detection chamber, the rotating stage drives the microfluidic luminescent detection reagent disk to rotate with the third rotation speed parameter, so that the cleaning solution is quantitatively released into the sample detection chamber; S6, the rotary table drives the microfluidic luminescent detection reagent disk to rotate with a fourth rotational speed parameter, so that the mixed liquid in the sample detection chamber is discharged into the waste liquid channel; S7, the rotating stage drives the microfluidic luminescent detection reagent disk to rotate with the fifth rotation speed parameter, so that the pre-excitation liquid is quantitatively released into the sample detection chamber and mixed with the reagent to be reacted for incubation; S8, the rotary stage drives the microfluidic luminescent detection reagent disk to rotate with the sixth rotation speed parameter, so that the excitation liquid is quantitatively released into the sample detection chamber to generate a light signal; S9, the optical signal is collected for component analysis.

2. The luminescence detection method according to claim 1, characterized in that: The pre-excitation liquid flow channel includes a pre-excitation liquid storage chamber, a pre-excitation liquid metering chamber, and a pre-excitation liquid quality control chamber. The pre-excitation liquid storage chamber, the pre-excitation liquid metering chamber, and the sample detection chamber are connected in series. The pre-excitation liquid quality control chamber is connected to the pre-excitation liquid metering chamber.

3. The luminescence detection method according to claim 1, characterized in that: The excitation fluid channel includes an excitation fluid storage chamber connected to the sample detection chamber.

4. The luminescence detection method according to claim 1, characterized in that: The reconstituted solution flow channel includes a reconstituted solution storage chamber, a reconstituted solution quantification chamber, and a reconstituted solution quality control chamber. The reconstituted solution storage chamber, the reconstituted solution quantification chamber, and the sample detection chamber are connected in series. The reconstituted solution quality control chamber is connected to the reconstituted solution quantification chamber.

5. The luminescence detection method according to claim 1, characterized in that: The cleaning fluid flow channel includes a cleaning fluid storage chamber, a cleaning fluid metering chamber, and a cleaning fluid quality control chamber. The cleaning fluid storage chamber, the cleaning fluid metering chamber, and the sample detection chamber are connected in series. The cleaning fluid quality control chamber is connected to the cleaning fluid metering chamber.

6. The luminescence detection method according to claim 5, characterized in that: Multiple cleaning fluid storage chambers are arranged in series, and multiple cleaning fluid quality control chambers are arranged in series. The number of cleaning fluid storage chambers is the same as the number of cleaning fluid quality control chambers.

7. The luminescence detection method according to claim 1, characterized in that: The sample liquid flow channel includes a plasma receiving chamber, a first plasma separation chamber, a second plasma separation chamber, and a plasma quality control chamber. The plasma receiving chamber, the first plasma separation chamber, and the second plasma separation chamber are connected in series. The sample detection chamber is connected in series between the first plasma separation chamber and the second plasma separation chamber. The plasma quality control chamber is connected to the first plasma separation chamber.

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