Microfluidic cartridge

Abstract

The application provides a microfluidic card box, which comprises a card box body and a reaction cup, and is characterized in that the card box body is provided with a fluid driving assembly, a fluid displacement chamber, a plurality of sample chambers and a reagent chamber; each sample chamber is provided with RPA reagents, the RPA reagents comprise RPA fluorescent probes, and the fluorescent groups marked by the RPA fluorescent probes in the sample chambers are different from each other and are used for distinguishing the sample chambers; and the fluid driving assembly is selectively connected with the sample chambers, the reagent chamber or the reaction cup through the fluid displacement chamber to drive liquid transmission. By arranging a plurality of independent sample chambers in a single card box body and marking the RPA fluorescent probes in the sample chambers with different fluorescent groups as molecular bar codes, a plurality of samples can be amplified in the same reaction cup, so that the single reaction can realize efficient detection of multiple samples, the throughput and efficiency of single detection are improved, and different samples from different sources can be accurately distinguished, and the accuracy and traceability of the detection result are improved.

Description

Technical Field

[0001] This invention relates to the field of molecular diagnostic technology, and in particular to a microfluidic cartridge. Background Technology

[0002] Molecular diagnostic technologies, particularly polymerase chain reaction (PCR) and isothermal amplification (AFA), have become crucial tools for pathogen detection. However, traditional PCR testing is highly dependent on specialized laboratories, expensive equipment, and highly trained personnel. The testing process is cumbersome and time-consuming (typically requiring -3 hours or even longer), and aerosol contamination during multi-step operations can lead to false negative results. These issues severely limit the widespread application of nucleic acid testing technology in point-of-care testing scenarios such as outpatient clinics, emergency rooms, customs, and primary healthcare facilities.

[0003] To address these issues, a fully automated molecular point-of-care testing (POCT) system integrating sample processing and amplification detection has emerged. This system integrates nucleic acid extraction, purification, and PCR detection using disposable microfluidic cartridges, achieving automated sample input and result output.

[0004] The applicant has discovered that the prior art has at least the following technical problems:

[0005] First, existing microfluidic cartridges are mostly designed for single-sample, single-test operation, which is insufficient to meet the needs of parallel testing of multiple samples in large-scale screening scenarios. When faced with scenarios requiring simultaneous testing of samples from multiple sources, multiple cartridges or multiple independent experiments are often required, resulting in high reagent costs and low testing efficiency. Furthermore, existing cartridges lack effective mechanisms for multi-sample differentiation and traceability, making it difficult to safely and accurately process multiple independent samples within a single cartridge.

[0006] Secondly, for samples with low nucleic acid load (such as tongue swab samples), since the samples may contain only extremely low concentrations of pathogens, the sensitivity of a single PCR amplification system is poor and there is a lack of design to verify the validity of the samples, which can easily lead to false negative results and seriously affect the accuracy of clinical diagnosis.

[0007] Therefore, designing a microfluidic cartridge with a simplified structure that can achieve parallel detection of multiple samples and independent traceability is a technical challenge that urgently needs to be solved in this field.

[0008] In view of this, the present invention is hereby proposed. Summary of the Invention

[0009] The purpose of this invention is to provide a microfluidic cartridge to address the design flaws of existing microfluidic cartridges, such as low detection throughput and difficulty in tracing multiple sample sources due to the single-sample, single-detection mode. The various technical effects of the preferred solutions provided by this invention are detailed below.

[0010] To achieve the above objectives, the present invention provides the following technical solution: This invention provides a microfluidic cartridge, comprising a cartridge body and a reaction cup. The cartridge body is provided with a fluid driving component, a fluid displacement chamber, multiple sample chambers, and a reagent chamber. Each sample chamber contains an RPA reagent, which includes an RPA fluorescent probe. The fluorescent groups labeled on the RPA fluorescent probes in each sample chamber are different to distinguish the sample chambers. The fluid driving component selectively connects the sample chambers, the reagent chambers, or the reaction cup through the fluid displacement chamber to drive liquid transport.

[0011] Preferably, the fluorescent group of the RPA fluorescent probe includes one of FAM, HEX, ROX, Cy5, or Cy5.5.

[0012] Preferably, at least one of the reagent chambers is provided with PCR reagents, the PCR reagents including a human internal reference gene fluorescent probe, the human internal reference gene fluorescent probe being labeled with the fluorescent group Atto425 at its 5' end and the quencher group BHQ1 at its 3' end.

[0013] Preferably, the reaction cup is provided with a single detection chamber, which is connected to the fluid inlet interface through a liquid inlet channel.

[0014] Preferably, the device further includes an optical detection device for acquiring fluorescence signals within the detection chamber of the reaction cup. The optical detection device includes an Atto425 channel, a FAM channel, a HEX channel, a ROX channel, a Cy5 channel, and a Cy5.5 channel to acquire the corresponding Atto425 fluorescence signal, FAM fluorescence signal, HEX fluorescence signal, ROX fluorescence signal, Cy5 fluorescence signal, and Cy5.5 fluorescence signal, respectively.

[0015] Preferably, the reaction cup is sealed to the cartridge body, and the fluid inlet of the reaction cup is in fluid communication with the fluid displacement chamber of the cartridge body.

[0016] Preferably, the cartridge body is further provided with an installation part and a first mating groove, and the reaction cup is further provided with an installation component, a fluid outlet port and a locking component. The installation component is adapted to the installation part, and the locking component is provided with a first snap-fit ​​part, which snaps into the first mating groove. The installation part is provided with a first sealing channel and a second sealing channel. The first sealing channel is sealed to the fluid inlet port through a first sealing component, and the second sealing channel is sealed to the fluid outlet port through a second sealing component.

[0017] Preferably, the fluid drive assembly includes a displacement turntable, a piston support, and a piston rod. The cartridge body has a receiving chamber, the piston support is rotatably disposed within the receiving chamber, and the piston support has a piston cavity. The piston rod reciprocates axially within the piston cavity. The displacement turntable is fixedly connected to the piston support. A sealing gasket is provided between the displacement turntable and the cartridge body. Through holes are provided at the bottom of the sample chamber and the reagent chamber. The sealing gasket has a connecting hole and a connecting channel. The connecting hole is aligned and communicates with the through hole, and the connecting channel is aligned and communicates with the first sealing channel. The fluid displacement chamber is disposed on the displacement turntable and communicates with the piston cavity, and is also connected to the connecting hole or the connecting channel via the displacement channel.

[0018] Preferably, the displacement turntable is provided with an annular protrusion, and the sealing gasket is provided with an annular sealing groove that seals with the annular protrusion to form a sealing area between the displacement turntable and the sealing gasket, and the displacement turntable and the sealing gasket are rotatably connected in a sealing manner; the opening of the displacement channel, the connecting hole, and the opening of the connecting channel are located within the sealing area.

[0019] Preferably, it also includes a base, the card box body is provided with a second snap-fit ​​portion, the base is provided with a second mating groove adapted to the second snap-fit ​​portion; the sealing gasket is provided with a third snap-fit ​​portion, the card box body is provided with a third mating groove adapted to the third snap-fit ​​portion; a receiving cavity is formed between the base and the card box body, and the replacement turntable is rotatably disposed in the receiving cavity.

[0020] The preferred technical solution of the present invention can also produce at least the following technical effects: The present invention provides a microfluidic cartridge, comprising a cartridge body and a reaction cup. The cartridge body is provided with a fluid driving component, a fluid displacement chamber, multiple sample chambers and a reagent chamber. Each sample chamber contains an RPA reagent, which includes an RPA fluorescent probe. The fluorescent groups labeled on the RPA fluorescent probes in each sample chamber are different to distinguish each sample chamber. The fluid driving component selectively connects the sample chambers, reagent chambers or reaction cups through the fluid displacement chamber to drive liquid transport.

[0021] This invention sets up multiple independent sample chambers within a single cartridge body, with each sample chamber's RPA fluorescent probe labeled with a different fluorescent group as a molecular barcode. Multiple samples can be amplified in the same reaction vessel, enabling efficient detection of multiple samples in a single reaction, improving throughput and efficiency. Furthermore, it can accurately distinguish samples from different sources, enhancing the accuracy and traceability of detection results. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0023] Figure 1 This is a schematic diagram of the structure of a microfluidic card holder provided by the present invention; Figure 2 This is a structural schematic diagram of a microfluidic card box provided by the present invention from another perspective; Figure 3 This is a cross-sectional view of a microfluidic card cartridge provided by the present invention; Figure 4 This is a schematic diagram of the structure of the card body and sealing gasket of a microfluidic card cartridge provided by the present invention; Figure 5 This is a schematic diagram of the structure of a microfluidic cartridge provided by the present invention, in which the reaction cup, the first sealing element, and the second sealing element are in a separated state; Figure 6 This is a schematic diagram of the displacement turntable and piston support of a microfluidic cartridge provided by the present invention; Figure 7 This is a schematic diagram of the structure of the receiving chamber, sample chamber, and reagent chamber of a microfluidic cartridge provided by the present invention.

[0024] In the picture: 1. Cartridge body; 11. Mounting part; 12. First mating groove; 13. First sealing channel; 14. Second sealing channel; 15. Second snap-fit ​​part; 16. Third mating groove; 17. Receiving chamber; 18. Sample chamber; 181. Sample injection port; 19. Sealing cap; 110. Reagent chamber; 1101. Cleaning chamber; 1102. Wetting chamber; 1103. Reconstituted solution chamber; 1104. Lyophilized bulb chamber; 111. Through hole; 2. Base; 21. Second mating groove; 22. Rotating groove; 3. Reaction cup; 31. Mounting component; 32. Locking component; 321. First snap-fit ​​part; 322. Fourth mating groove; 33. Fluid inlet port; 34. Fluid outlet port; 35. Venting groove; 36. Inlet channel; 361. Inlet bend section; 37. Outlet channel; 371. Outlet bend section; 38. Detection chamber; 4. Displacement turntable; 41. Displacement channel; 42. Fluid displacement chamber; 43. Annular protrusion; 44. Grip protrusion; 45. Indicator mark; 46. Rotating ring; 5. Piston support; 51. Piston chamber; 6. Sealing gasket; 61. Communicating hole; 62. Communicating channel; 63. Annular sealing groove; 64. Third snap-fit ​​part; 7. First sealing element; 8. Second sealing element; 9. Outer shell; 91. Fourth snap-fit ​​part. Detailed Implementation

[0025] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be described in detail below. Obviously, the described embodiments are merely some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other implementation methods obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0026] like Figures 1 to 7 As shown, the present invention provides a microfluidic cartridge, including a cartridge body and a reaction cup. The cartridge body is provided with a fluid driving component, a fluid displacement chamber, multiple sample chambers and a reagent chamber. Each sample chamber contains an RPA reagent, which includes an RPA fluorescent probe. The fluorescent groups labeled on the RPA fluorescent probes in each sample chamber are different to distinguish each sample chamber. The fluid driving component selectively connects the sample chambers, reagent chambers or reaction cups through the fluid displacement chamber to drive liquid transport.

[0027] This invention sets up multiple independent sample chambers 18 within a single cartridge body 1, and each sample chamber 18 has an RPA fluorescent probe labeled with a different fluorescent group as a molecular barcode. Multiple samples can be amplified in the same reaction cup 3. This not only enables efficient detection of multiple samples in a single reaction, improving the throughput and efficiency of a single detection, but also accurately distinguishes samples from different sources, improving the accuracy and traceability of the detection results.

[0028] Furthermore, each sample chamber 18 has an independent sample injection port 181, and the sample injection port 181 is equipped with a sealing cap 19. The sealing cap 19 has an independent QR code, the encoded information of which includes the sample chamber 18's number, batch number, and serial number. In practical applications, by scanning the QR code on the sealing cap 19, the encoded information is read and bound to the source information of the sample to be tested in the corresponding information system, achieving a one-to-one correspondence between the sample and the sample chamber 18. Subsequently, the sealing cap 19 is opened, and the sample to be tested is placed into the corresponding sample chamber 18 through the sample injection port 181. Because each sample chamber 18 is independent, complete isolation between different samples is achieved, avoiding cross-contamination between samples.

[0029] As an optional implementation, the fluorescent group of the RPA fluorescent probe includes one of FAM, HEX, ROX, Cy5, or Cy5.5.

[0030] As an optional implementation, at least one reagent chamber is provided with PCR reagents, which include a human internal reference gene fluorescent probe. The human internal reference gene fluorescent probe is labeled with the fluorescent group Atto425 at its 5' end and with the quencher group BHQ1 at its 3' end.

[0031] As an optional implementation, an optical detection device is also included, which is used to acquire fluorescence signals within the detection chamber 38 of the reaction cup 3. The optical detection device includes an Atto425 channel, a FAM channel, a HEX channel, a ROX channel, a Cy5 channel, and a Cy5.5 channel to acquire the corresponding Atto425 fluorescence signal, FAM fluorescence signal, HEX fluorescence signal, ROX fluorescence signal, Cy5 fluorescence signal, and Cy5.5 fluorescence signal, respectively.

[0032] During RPA pre-amplification, the RPA probes in each sample chamber 18 are pre-labeled with distinct fluorescent groups as molecular barcodes. During pre-amplification, these fluorescent groups integrate with the RPA probes into their respective amplification products and are retained during PCR amplification. Therefore, the source of different samples can be distinguished through the fluorescence channel, enabling independent traceability of results. Compared to traditional tube-based testing methods, this invention eliminates the need for separate reaction containers for each sample, simplifying the cartridge structure and avoiding the risk of cross-contamination from multiple pipetting operations. Furthermore, the independent QR code on each sealing cap 19 further strengthens the binding between physical samples and digital information, achieving comprehensive traceability.

[0033] A human internal reference gene fluorescent probe was placed in the PCR reagent in reagent chamber 110 to verify the validity of the sample itself, i.e., to confirm whether the sample contains a sufficient number of human cells. During result interpretation, the Atto425 channel needs to be calibrated first. When there is no signal in the Atto425 channel, the sample test result is considered invalid, thereby improving the accuracy and reliability of the test results.

[0034] A two-stage strategy combining RPA pre-amplification and PCR precision detection was employed. RPA pre-amplification amplifies the trace target nucleic acid signal by 100–1000 times, addressing the sensitivity issue of low bacterial load samples such as tongue swabs. Simultaneously, the distinct fluorescent groups on the specific RPA fluorescent probes in each sample chamber are integrated into the pre-amplification product. Subsequent PCR precision detection utilizes its high specificity to filter out non-specific products that may arise during the RPA pre-amplification stage, improving the accuracy and reliability of the results.

[0035] During result interpretation, since the pre-amplified products from each sample chamber are collected in reaction cup 3, the optical detection device captures the initial fluorescence value released by the pre-amplified products at the beginning of the PCR reaction and uses this as a baseline. In the subsequent PCR amplification stage, if the channel corresponding to a sample exhibits exponential amplification and its cycle threshold (Ct value) reaches the preset positive determination threshold, the sample is determined to be positive; otherwise, the sample is determined to be negative.

[0036] As an optional implementation, such as Figure 3 , Figure 4 , Figure 5As shown, the reaction cup 3 is sealed to the cartridge body 1, and the fluid inlet 33 of the reaction cup 3 is fluidly connected to the fluid replacement chamber 42. The cartridge body 1 is also provided with an installation part 11 and a first mating groove 12. The reaction cup 3 is also provided with an installation part 31, a fluid outlet 34, and a locking part 32. The installation part 31 is adapted to the installation part 11. The locking part 32 is provided with a first snap-fit ​​part 321, and the snap-fit ​​structure of the first snap-fit ​​part 321 snaps into the first mating groove 12. The installation part 11 is provided with a first sealing channel 13 and a second sealing channel 14. The first sealing channel 13 is sealed to the fluid inlet 33 through a first sealing element 7, and the second sealing channel 14 is sealed to the fluid outlet 34 through a second sealing element 8.

[0037] Furthermore, the first seal 7 adopts a sealing ring structure. The second seal 8 has a structure that is closed at one end and open at the other. The mounting part 11 has a groove structure, and the mounting part 31 is engaged with the mounting part 11.

[0038] When the reaction cup 3 is installed on the cartridge body 1, the locking member 32 engages with the first mating groove 12 through the first snap-fit ​​part 321 to apply a clamping force, so that the mounting member 31 and the mounting part 11 are tightly abutted. At this time, the first sealing channel 13 is sealed to the fluid inlet interface 33 through the first sealing member 7, and the second sealing channel 14 is sealed to the fluid outlet interface 34 through the second sealing member 8, thereby improving the overall sealing performance of the reaction cup 3 and the cartridge body 1.

[0039] The reaction cup 3 is equipped with an independent detection chamber 38. After the fluid enters from the fluid inlet port 33, it flows into the bottom of the detection chamber 38 through the inlet channel 36. After the reaction is completed, it is collected through the outlet channel 37 and discharged to the fluid outlet port 34.

[0040] The outlet of the liquid inlet channel 36 and the inlet of the liquid outlet channel 37 are positioned opposite each other on the detection chamber 38, so that under the action of the fluid drive component, the liquid will be pushed upward from the bottom of the detection chamber 38, gradually filling the entire chamber space, and finally discharged from the top of the detection chamber 38.

[0041] The main body of the cartridge 1 is made of medical-grade polypropylene. The reaction cup 3 is a separately molded, optically transparent cup.

[0042] It also includes a housing 9, which has a fourth snap-fit ​​part 91, and a locking member with a fourth mating groove 322 that is adapted to the fourth snap-fit ​​part 91. Through the snap-fit ​​engagement of the fourth snap-fit ​​part 91 and the fourth mating groove 322, the housing 9 is fitted over the outside of the reaction cup 3, providing a certain degree of protection.

[0043] As an optional implementation, such as Figure 3 , Figure 5As shown, the reaction cup 3 is provided with a single detection chamber 38, which is connected to the fluid inlet interface 33 through the liquid inlet channel 36.

[0044] Furthermore, a one-way valve is provided at the inlet of the fluid inlet interface 33. The one-way valve is used to allow liquid to flow only from the fluid drive component to the detection chamber 38. The one-way valve includes a miniature passive one-way valve, which restricts the flow of liquid to the detection chamber 38 only from the fluid displacement chamber 42, effectively preventing liquid backflow and eliminating the possibility of liquid or gas flowing in reverse or between chambers through the flow channel.

[0045] The inner diameter of the fluid inlet port 33 gradually decreases along the liquid flow direction; the outlet of the fluid inlet port 33 is connected to the inlet of the inlet channel 36 through the venting groove 35. The venting groove 35 is used to trap and store some of the gas carried in the liquid flowing into the fluid inlet port 33. The cross-section of the venting groove 35 is trapezoidal, and its inner diameter gradually decreases along the liquid flow direction. The increased cross-section reduces the flow velocity, promoting gas to rise and separate. The minimum inner diameter of the venting groove 35 is larger than the inner diameter of the inlet of the inlet channel 36.

[0046] The tapered inner diameter design of the fluid inlet port 33 allows liquid to be injected into the exhaust groove 35 quickly, and the kinetic energy of the fluid is used to quickly push out some of the gas and temporarily store it in the exhaust groove 35.

[0047] Through the synergistic effect of the detection chamber 38, fluid inlet interface 33, fluid outlet interface 34, inlet channel 36, outlet channel 37 and exhaust groove 35, the retention and generation of bubbles are effectively reduced, the interference of bubble structure on optical detection signal is avoided, and the stability of detection is improved.

[0048] The exhaust channel 35, the liquid inlet channel 36, the detection chamber 38, and the liquid outlet channel 37 are each sealed on both sides by an encapsulation membrane. Micro-channels are provided on the encapsulation membrane within the detection chamber 38. These micro-channels are arranged in a nested, radial, or grid pattern on the encapsulation membrane. When liquid enters from the bottom of the detection chamber 38, the micro-channels guide the liquid to spread evenly in all directions, allowing the liquid to quickly and completely cover the entire detection chamber 38, preventing the liquid from flowing only along the center and leaving dry areas around the perimeter. It should be noted that the term "micro-channels" refers to channels with a structural size at the micrometer level.

[0049] Both the inlet channel 36 and the outlet channel 37 are provided with at least one curved section, and the curved section adopts a rounded transition structure. The curved section of the rounded transition structure is C-shaped, U-shaped, or S-shaped. The inlet channel 36 is provided with an inlet curved section 361, and the outlet channel 37 is provided with an outlet curved section 371. This design not only effectively extends the flow path of the liquid, but also avoids the formation of flow dead zones at right-angle bends, reduces local resistance loss of the fluid, and effectively reduces the non-specific adsorption of biological samples such as proteins at sharp corners, further improving the accuracy of the detection results.

[0050] As an optional implementation, such as Figure 3 , Figure 4 , Figure 5 , Figure 6 As shown, the fluid drive assembly includes a displacement turntable 4, a piston support 5, and a piston rod. A receiving chamber 17 is provided on the cartridge body 1. The piston support 5 is rotatably disposed within the receiving chamber 17 and has a piston cavity 51. The piston rod reciprocates axially within the piston cavity 51. The displacement turntable 4 is fixedly connected to the piston support 5. A sealing gasket 6 is provided between the displacement turntable 4 and the cartridge body 1. Each sample chamber 18 and reagent chamber 110 has an independent through hole 111 at its bottom. The sealing gasket 6 has multiple connecting holes 61111 and connecting channels 62. Each connecting hole 61111 corresponds to and is connected to each through hole 111. The connecting channel 62 is aligned and connected to the first sealing channel 13. A fluid displacement chamber 42 is disposed on the displacement turntable 4 and is connected to the piston cavity 51. The fluid displacement chamber 42 is connected to the connecting hole 61111 or the connecting channel 62 through the displacement channel 41.

[0051] Furthermore, a gripping protrusion 44 is provided on the bottom surface of the replacement turntable 4 to facilitate the user's gripping and rotation of the replacement turntable 4, making the rotation operation more effortless and smooth.

[0052] There are five sample chambers 18, and each sample chamber 18 can be evenly arranged around the outer periphery of the receiving chamber 17, or arranged side by side on the main body of the card box 1.

[0053] The bottom surface of the replacement turntable 4 is also equipped with an indicator 45, which corresponds to the replacement channel 41. This provides users with intuitive visual guidance, enables accurate positioning, and reduces the risk of misoperation.

[0054] An external power source drives the piston head of the piston rod to reciprocate linearly within the piston chamber 51, thereby changing the volume and pressure within the piston chamber 51. Since the piston chamber 51 is connected to the fluid replacement chamber 42, the liquid in the reagent chamber 110 is drawn into the fluid replacement chamber 42 through the replacement channel 41, or the liquid in the fluid replacement chamber 42 is pushed into the corresponding reagent chamber 110 or detection chamber 38.

[0055] Specifically, an external power source drives the piston rod to reciprocate axially within the piston chamber 51. The retraction of the piston rod increases the volume of the piston chamber 51, creating a negative pressure in the fluid replacement chamber 42. Under the pressure difference, the liquid in the corresponding sample chamber 18 or reagent chamber 110 is sequentially drawn into the fluid replacement chamber 42 through its through-hole 111, the corresponding connecting hole 61111 on the sealing gasket 6, and the replacement channel 41 on the replacement turntable 4. Conversely, the advancement of the piston rod decreases the volume of the piston chamber 51, creating a positive pressure in the fluid replacement chamber 42. The liquid is pushed back to the corresponding reagent chamber 110 through the replacement channel 41, the corresponding connecting hole 61111, and the corresponding through-hole 111, or pushed to the fluid inlet port 33 through the replacement channel 41, the corresponding connecting channel 62 on the sealing gasket 6, and the first sealing channel 13, and then enters the detection chamber 38 under the guidance of the one-way valve. The fluid replacement chamber 42 is connected to the sample chamber 18 and the reagent chamber 110 through the alignment of the connecting hole 61111 and the through hole 111 to achieve bidirectional flow. The fluid replacement chamber 42 and the detection chamber 38 are connected through a one-way valve to allow only unidirectional flow from the fluid replacement chamber 42 to the detection chamber 38, which effectively prevents liquid or gas in the detection chamber 38 from flowing back to the fluid replacement chamber 42 and avoids cross-contamination of samples during multi-channel parallel detection.

[0056] As an optional implementation, such as Figure 3 , Figure 4 , Figure 6 As shown, the displacement turntable 4 is provided with an annular protrusion 43, and the sealing gasket 6 is provided with an annular sealing groove 63 that seals with the annular protrusion 43 to form a sealing area between the displacement turntable 4 and the sealing gasket 6, and the displacement turntable 4 and the sealing gasket 6 are rotatably connected in a sealed manner; the opening of the displacement channel 41, the opening of the connecting hole 61111, and the opening of the connecting channel 62 are located within the sealing area.

[0057] As an optional implementation, such as Figure 2 , Figure 3 As shown, it also includes a base 2, a second latching part 15 is provided on the card box body 1, and a second mating groove 21 adapted to the second latching part 15 is provided on the base 2; a third latching part 64 is provided on the sealing gasket 6, and a third mating groove 16 adapted to the third latching part 64 is provided on the card box body 1; a receiving cavity is formed between the base 2 and the card box body 1, and the replacement turntable 4 is rotatably disposed in the receiving cavity.

[0058] Furthermore, the base 2 has an annular rotating groove 22, and the replacement turntable 4 is provided with a rotating ring 46 that matches the annular rotating groove 22. Through the rotatable engagement of the rotating ring 46 and the annular rotating groove 22, the replacement turntable 4 is rotatably set in the receiving space.

[0059] When the user rotates the replacement turntable 4, the annular protrusion 43 on the replacement turntable 4 and the annular sealing groove 63 on the sealing gasket 6 remain in close contact, so that dynamic sealing is achieved during the process of the replacement turntable 4 rotating to switch the flow path.

[0060] The user rotates the replacement turntable 4 by holding the grip protrusion 44, so that the opening of the replacement channel 41 on the replacement turntable 4 is aligned with the different connecting holes 61111 or connecting channels 62 on the sealing gasket 6 in sequence, thereby switching the target chamber connected to the fluid replacement chamber 42; the indicator mark 45 on the bottom surface of the replacement turntable 4 provides visual guidance for the flow path switching.

[0061] During the rotation of the displacement turntable 4, the annular protrusion 43 on the displacement turntable 4 always maintains a tight fit with the annular sealing groove 63 on the sealing gasket 6, achieving dynamic sealing under rotation conditions. The base 2 is engaged with the second mating groove 21 of the cartridge body 1 through the hook structure of the second engaging part 15, and the sealing gasket 6 is engaged with the third mating groove 16 of the cartridge body 1 through the hook structure of the third engaging part 64. The three together define the accommodating space for the sealing gasket 6 and the displacement turntable 4, improving the structural stability and sealing reliability of the fluid drive assembly when rotating and switching flow paths.

[0062] As an optional implementation, such as Figure 7 As shown, reagent chamber 110 includes a washing chamber 1101, a rinsing chamber 1102, a reconstitution chamber 1103, and a lyophilized bulb chamber 1104, with PCR reagent pre-filled in the lyophilized bulb chamber 1104. The washing chamber 1101 is used to wash away impurities during nucleic acid extraction. The rinsing chamber 1102 is used to remove residual washing solution from the washing chamber 1101. The reconstitution chamber 1103 is used to elute and dissolve the purified nucleic acid. The lyophilized PCR reagent in the lyophilized bulb chamber 1104 is rapidly activated after reconstitution.

[0063] Furthermore, five sample chambers 18, along with a washing chamber 1101, a rinsing chamber 1102, a reconstitution chamber 1103, and a lyophilized bulb chamber 1104, are arranged sequentially around the containing chamber 17. Each sample chamber 18 and each reagent chamber 110 is independently configured, effectively avoiding cross-contamination between samples and reagents. An air gap is provided between adjacent chambers. The thickness of the air gap is 0.5mm-2mm, utilizing the property of air as a poor conductor of heat to form a physical barrier and thermal insulation barrier between the chambers.

[0064] The multi-chamber microfluidic cartridge provided by this invention can simultaneously detect tongue swab samples from 5 different individuals in a single run on existing improved instruments. It features short detection time, high sensitivity, good specificity, no cross-contamination, and controllable per capita consumable costs. It is especially suitable for promotion and application in large-scale single-target multi-sample screening in primary healthcare units and communities.

[0065] It should be noted that the specific type, quantity, and specifications of reagent chamber 110 can be flexibly selected according to usage requirements.

[0066] It is understood that the same or similar parts in the above embodiments can be referred to each other, and the contents not described in detail in some embodiments can be referred to the same or similar contents in other embodiments.

[0067] In the description of this invention, it should be noted that, unless otherwise stated, "a plurality of" means two or more; the terms "upper," "lower," "left," "right," "inner," "outer," "front end," "rear end," "head," "tail," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, and are only for the convenience of describing the invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of the invention. Furthermore, the terms "first," "second," "third," etc., are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0068] In the description of this invention, it should also be noted that, unless otherwise explicitly specified and limited, the terms "installation," "connection," and "joining" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0069] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "a particular example," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of this application. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0070] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A microfluidic card cartridge, characterized in that, The device includes a cartridge body and a reaction cup. The cartridge body is provided with a fluid driving component, a fluid displacement chamber, multiple sample chambers, and a reagent chamber. Each sample chamber contains an RPA reagent, which includes an RPA fluorescent probe. The fluorescent groups labeled on the RPA fluorescent probes in each sample chamber are different to distinguish each sample chamber. The fluid driving component selectively connects the sample chambers, the reagent chambers, or the reaction cup through the fluid displacement chamber to drive liquid transport.

2. A microfluidic card holder according to claim 1, characterized in that, The fluorescent group of the RPA fluorescent probe includes one of FAM, HEX, ROX, Cy5, or Cy5.

5.

3. A microfluidic card holder according to claim 2, characterized in that, At least one of the reagent chambers is provided with PCR reagents, the PCR reagents including a human internal reference gene fluorescent probe, the human internal reference gene fluorescent probe being labeled with the fluorescent group Atto425 at its 5' end and the quencher group BHQ1 at its 3' end.

4. A microfluidic card holder according to claim 1, characterized in that, The reaction cup is equipped with a single detection chamber, which is connected to the fluid inlet interface via a liquid inlet channel.

5. A microfluidic card holder according to claim 3, characterized in that, It also includes an optical detection device, which is used to collect fluorescence signals in the detection chamber of the reaction cup. The optical detection device includes an Atto425 channel, a FAM channel, a HEX channel, a ROX channel, a Cy5 channel, and a Cy5.5 channel to collect the corresponding Atto425 fluorescence signal, FAM fluorescence signal, HEX fluorescence signal, ROX fluorescence signal, Cy5 fluorescence signal, and Cy5.5 fluorescence signal, respectively.

6. A microfluidic card holder according to claim 1, characterized in that, The reaction cup is sealed to the cartridge body, and the fluid inlet of the reaction cup is in fluid communication with the fluid displacement chamber of the cartridge body.

7. A microfluidic card holder according to claim 6, characterized in that, The cartridge body is also provided with an installation part and a first mating groove. The reaction cup is also provided with an installation component, a fluid outlet port and a locking component. The installation component is adapted to the installation part. The locking component is provided with a first snap-fit ​​part, which snaps into the first mating groove. The installation part is provided with a first sealing channel and a second sealing channel. The first sealing channel is sealed to the fluid inlet port through a first sealing component, and the second sealing channel is sealed to the fluid outlet port through a second sealing component.

8. A microfluidic card holder according to claim 7, characterized in that, The fluid drive assembly includes a displacement turntable, a piston support, and a piston rod. The cartridge body has a receiving chamber, and the piston support is rotatably disposed within the receiving chamber. The piston support has a piston cavity, and the piston rod reciprocates axially within the piston cavity. The displacement turntable is fixedly connected to the piston support. A sealing gasket is provided between the displacement turntable and the cartridge body. Through holes are provided at the bottom of the sample chamber and the reagent chamber. The sealing gasket has a connecting hole and a connecting channel. The connecting hole is aligned and communicates with the through hole, and the connecting channel is aligned and communicates with the first sealing channel. The fluid displacement chamber is disposed on the displacement turntable and communicates with the piston cavity, and is also connected to the connecting hole or the connecting channel via the displacement channel.

9. A microfluidic card holder according to claim 8, characterized in that, The displacement turntable is provided with an annular protrusion, and the sealing gasket is provided with an annular sealing groove that seals with the annular protrusion to form a sealing area between the displacement turntable and the sealing gasket, and the displacement turntable and the sealing gasket are rotatably connected in a sealing manner; the opening of the displacement channel, the connecting hole and the opening of the connecting channel are located within the sealing area.

10. A microfluidic card cartridge according to claim 9, characterized in that, It also includes a base, a second latching part is provided on the card box body, and a second mating groove adapted to the second latching part is provided on the base; a third latching part is provided on the sealing gasket, and a third mating groove adapted to the third latching part is provided on the card box body; a receiving cavity is formed between the base and the card box body, and the replacement turntable is rotatably disposed in the receiving cavity.

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