POCT cartridge with nucleic acid extraction cavity outside

Abstract

This invention provides a POCT cartridge with an externally mounted nucleic acid extraction chamber, comprising: a reagent chamber body, which has a plunger cavity extending vertically through both ends and multiple receiving cavities surrounding the plunger cavity, with a plunger located in the top opening of each plunger cavity; and a nucleic acid extraction chamber, formed on and protruding from the outer wall of the reagent chamber body, the internal space of which is controllably connected to the plunger cavity, containing a magnetic bead, and having a receiving groove on the bottom wall of the chamber for accommodating the ultrasonic head of an ultrasonic device. This invention, by having the nucleic acid extraction chamber protrude independently from the outer wall of the reagent chamber body, allows for independent structural design of the nucleic acid extraction chamber. The receiving groove on its bottom wall reliably matches the ultrasonic head, enabling efficient mixing of the sample liquid using ultrasound, thereby improving the efficiency of the entire nucleic acid extraction and detection process.

Description

Technical Field

[0001] This invention belongs to the field of point-of-care testing technology, specifically relating to a POCT cartridge with an external nucleic acid extraction chamber. Background Technology

[0002] PCR (Polymerase Chain Reaction) is a molecular biology technique that amplifies specific DNA (deoxyribonucleic acid) sequences in vitro. Due to its high specificity, high sensitivity, low purity requirements, and ease of use and speed, PCR is widely used in molecular biology detection and analysis. Routine nucleic acid testing requires a PCR laboratory. According to national regulations, PCR laboratories must be divided into separate zones: reagent preparation, nucleic acid extraction, amplification, and detection. Furthermore, relevant personnel must possess PCR certification. These regulations impose certain requirements on both the experimental environment and the personnel's qualifications. However, even with these stringent requirements, aerosol contamination can still affect the accuracy of test results.

[0003] To prevent the adverse effects of external aerosol contamination on test results, existing technologies for nucleic acid testing cartridges use multiple reagent chambers to transfer samples and reagents via multiple on / off valves. However, this method hinders the miniaturization of the device and cartridge. Patent application number 2022111389126 proposes a nucleic acid testing cartridge that uses plunger suction and a rotary valve to select different chambers, enabling the mixing and extraction of nucleic acid samples within each chamber. However, this reciprocating suction method is less efficient than ultrasonic mixing, reducing the efficiency of point-of-care nucleic acid extraction and testing. Especially compared to membrane-based nucleic acid extraction, magnetic bead extraction requires even higher mixing precision. Therefore, designing a POCT cartridge capable of ultrasonic mixing is a pressing issue. Summary of the Invention

[0004] This invention provides a POCT cartridge with an externally mounted nucleic acid extraction chamber, which solves the technical problem that existing nucleic acid detection cartridges are not suitable for ultrasonic mixing, resulting in poor mixing of nucleic acid samples and low efficiency of extraction and detection, especially unsuitable for magnetic bead extraction of nucleic acid.

[0005] To address the above problems, the present invention provides a POCT cartridge with an externally mounted nucleic acid extraction cavity, comprising:

[0006] The reagent chamber body has a plunger cavity that extends vertically through both ends and a plurality of receiving cavities arranged around the plunger cavity. A plunger is provided in the top opening of the plunger cavity.

[0007] A nucleic acid extraction cavity is formed on the outer wall of the reagent cavity body and protrudes from the outer wall of the reagent cavity body. The internal space of the nucleic acid extraction cavity can be controllably connected with the plunger cavity. The nucleic acid extraction cavity contains a magnetic bead. A receiving groove is constructed on the bottom wall of the nucleic acid extraction cavity for receiving the ultrasonic head of the ultrasonic device.

[0008] In some embodiments, the outer wall of the nucleic acid extraction cavity is connected to the outer wall of the reagent cavity body by a snap-fit ​​connection; and / or, it further includes a flat reaction cavity formed on the outer wall of the reagent cavity body and protruding from the outer wall of the reagent cavity body, wherein the internal space of the flat reaction cavity can be controllably communicated with the plunger cavity.

[0009] In some embodiments, the external POCT cartridge of the nucleic acid extraction chamber further includes: a rotary valve, including a valve body and a protrusion on one side of the valve body, the protrusion being inserted into the bottom opening of the plunger cavity, a flow channel being constructed inside the rotary valve, a first port of the flow channel being constructed on the top surface of the valve body, a second port of the flow channel being constructed on the free end face of the protrusion, and the second port extending along the axial direction of the protrusion, and a chamber inlet / outlet being constructed on the bottom wall of each of the accommodating cavities, and a first reaction chamber inlet / outlet communicating with the internal space of the flat reaction chamber and a first extraction chamber inlet / outlet communicating with the internal space of the nucleic acid extraction chamber being constructed on the bottom end face of the reagent chamber body, when the rotary valve is driven to rotate, the first port can be selectively aligned with one of the chamber inlet / outlet, the first reaction chamber inlet / outlet, and the first extraction chamber inlet / outlet, so that the liquid flowing through the inlet / outlet aligned with the first port can directly communicate with the plunger cavity through the flow channel.

[0010] In some embodiments, the inlet and outlet ports of each of the chambers, the inlet and outlet ports of the first reaction chamber, and the inlet and outlet ports of the first extraction chamber are located on a first circle, the first circle being coaxial with the protrusion, and the first port being located on the first circle; and / or, a wedge-shaped groove is constructed on the end face of the valve body away from the reagent chamber body.

[0011] In some embodiments, the external POCT cartridge of the nucleic acid extraction chamber further includes: a base, the base being detachably assembled to the bottom end of the reagent chamber body, and an assembly gap being formed between the base and the bottom end of the reagent chamber body, the rotary valve being rotatably connected within the assembly gap, and the bottom surface of the valve body being flush with the bottom surface of the base.

[0012] In some embodiments, the base includes a base body, the central region of the top surface of the base body having a connecting protrusion extending toward the reagent chamber body, the connecting protrusion being snapped together with the bottom end of the reagent chamber body, and the top surface of the connecting protrusion being supported below the bottom end surface of the valve body.

[0013] In some embodiments, the outer wall of the flat reaction chamber is detachably connected to the outer wall of the reagent chamber body.

[0014] In some embodiments, the top surface of the base body is provided with an anti-detachment structure extending toward the reagent chamber body. The anti-detachment structure includes spaced and oppositely arranged bent baffles. The amplification chamber of the flat reaction chamber extends out of the gap between the two bent baffles. The flat reaction chamber has a connection and mating part with the reagent chamber body. The connection and mating part is clamped between the bent baffles and the outer wall of the reagent chamber body.

[0015] In some embodiments, the POCT cartridge external to the nucleic acid extraction chamber further includes:

[0016] The first sealing gasket is circular and is sandwiched between the top surface of the valve body and the bottom surface of the reagent chamber body. The first sealing gasket has liquid passage holes corresponding to the liquid inlet and outlet of each of the chambers, the liquid inlet and outlet of the first reaction chamber, the liquid inlet and outlet of the first extraction chamber, and the first port, respectively, as well as perforations for passing through the protrusion.

[0017] In some embodiments, a plurality of first positioning posts and second positioning posts are formed on the bottom end face of the reagent chamber body. The first sealing gasket has a first positioning through hole corresponding to each of the first positioning posts and a second positioning through hole corresponding to each of the second positioning posts. The first positioning posts are evenly spaced along the outer circumference of the first sealing gasket, and the second positioning posts are evenly spaced around the through holes and located radially inside the first sealing gasket; and / or, the bottom end face of the reagent chamber body is integrally injection molded with the first sealing gasket.

[0018] The present invention provides a POCT cartridge with an externally mounted nucleic acid extraction cavity, which has the following beneficial effects:

[0019] The nucleic acid extraction chamber protrudes independently from the outer wall of the reagent chamber body, instead of surrounding the plunger cavity with the aforementioned accommodating cavities. This allows for independent structural design of the nucleic acid extraction chamber. The accommodating groove on its bottom wall reliably matches the ultrasonic head, enabling efficient ultrasonic mixing of the sample liquid within. This improves the efficiency of the entire nucleic acid extraction and detection process. Furthermore, because the nucleic acid extraction chamber protrudes from the outer wall of the reagent chamber body, the distance between it and the accommodating cavities within the reagent chamber body is increased, effectively reducing adverse interference from the ultrasonic device to reagents or liquids in other adjacent accommodating cavities. Additionally, the protruding nucleic acid extraction chamber facilitates easier matching with the magnetic bead adsorption device, enabling effective adsorption of the magnetic beads. Attached Figure Description

[0020] To more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings in the following description are merely exemplary, and those skilled in the art can derive other embodiments based on the provided drawings without creative effort.

[0021] The structures, proportions, sizes, etc. illustrated in this specification are only for the purpose of assisting those skilled in the art in understanding and reading the content disclosed herein, and are not intended to limit the conditions under which the present invention can be implemented. Therefore, they have no substantial technical significance. Any modifications to the structure, changes in the proportions, or adjustments to the size, without affecting the effects and objectives that the present invention can produce, should still fall within the scope of the technical content disclosed in the present invention.

[0022] Figure 1 This is an exploded view of the structure of the POCT cartridge externally mounted to the nucleic acid extraction chamber in an embodiment of the present invention;

[0023] Figure 2 This is a front view of the external POCT card holder of the nucleic acid extraction chamber according to an embodiment of the present invention;

[0024] Figure 3 for Figure 1 A front view of the top cover (open state);

[0025] Figure 4 for Figure 3 Top view;

[0026] Figure 5 for Figure 1 Top view of the second sealing gasket in the middle;

[0027] Figure 6 for Figure 1A top view of the reagent chamber body;

[0028] Figure 7 for Figure 6 Side view of the nucleic acid extraction chamber;

[0029] Figure 8 for Figure 6 Side view of the flat reaction chamber;

[0030] Figure 9 for Figure 1 A bottom view of the reagent chamber body;

[0031] Figure 10 for Figure 1 A top view of the rotary valve in the image;

[0032] Figure 11 for Figure 1 A bottom view of the rotary valve in the image;

[0033] Figure 12 for Figure 1 Top view of the first sealing gasket in the middle;

[0034] Figure 13 for Figure 1 Side view of the nucleic acid extraction chamber in the middle;

[0035] Figure 14 for Figure 1 A three-dimensional structural diagram of the base;

[0036] Figure 15 for Figure 1 A schematic diagram of the exploded structure of the flat reaction chamber in the image;

[0037] Figure 16 for Figure 1 A schematic diagram of a flat reaction chamber (the first and second membranes are omitted).

[0038] Figure 17 for Figure 1 Another schematic diagram of the flat reaction chamber (the first and second membranes are omitted).

[0039] The reference numerals in the attached figures are as follows:

[0040] 1. Reagent chamber body; 11. Plunger chamber; 12. Chamber inlet / outlet; 131. First positioning column; 132. Second positioning column; 141. Sample chamber; 142. Waste liquid chamber; 143. Proteinase K chamber; 144. Elution buffer chamber; 1451. First washing chamber; 1452. Second washing chamber; 1453. Third washing chamber; 1454. Fourth washing chamber; 1455. Fifth washing chamber; 145. Lyophilized bulb chamber; 1461. Reaction chamber connection hole; 1462. Extraction chamber connection hole; 2. Nucleic acid extraction chamber; 21. Buckle; 22. First extraction chamber inlet / outlet; 23. Sealing ring; 3. Plunger; 4. Flat reaction chamber; 41. First reaction chamber inlet / outlet; 42. Connecting mating part; 431. Second reaction chamber inlet / outlet ; 432, First outlet; 441, Main flow channel; 442, Branch flow channel; 45, Amplification chamber; 451, Amplification chamber vent; 461, First diaphragm; 462, Second diaphragm; 47, Resistance section; 5, Rotary valve; 51, Valve body; 511, First port; 512, Second port; 513, Wedge groove; 52, Protrusion; 6, Base; 61, Base body; 62, Connecting protrusion; 621, First locking protrusion; 63, Bending baffle; 7, First sealing gasket; 71, Liquid passage hole; 72, Perforation; 731, First positioning through hole; 732, Second positioning through hole; 8, Top cover; 81, Cover body; 811, Vent hole; 812, Sample loading hole; 813, Second locking protrusion; 814, Plunger through hole; 82, Cap; 9, Second sealing gasket. Detailed Implementation

[0041] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. The following description of at least one exemplary embodiment is merely illustrative and is in no way intended to limit the present invention or its application or use. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0042] It should be noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the exemplary embodiments according to this application. As used herein, the singular form is intended to include the plural form as well, unless the context clearly indicates otherwise. Furthermore, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, devices, components, and / or combinations thereof.

[0043] It should be understood that the term "and / or" used in this article is merely a description of the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can represent: A existing alone, A and B existing simultaneously, and B existing alone. Additionally, the character " / " in this article generally indicates that the preceding and following related objects have an "or" relationship.

[0044] Unless otherwise specifically stated, the relative arrangement, numerical expressions, and values ​​of the components and steps set forth in these embodiments do not limit the scope of the invention. It should also be understood that, for ease of description, the dimensions of the various parts shown in the drawings are not drawn to actual scale. Techniques, methods, and devices known to those skilled in the art may not be discussed in detail, but where appropriate, such techniques, methods, and devices should be considered part of the specification. In all examples shown and discussed herein, any specific values ​​should be interpreted as merely exemplary and not as limitations. Therefore, other examples of exemplary embodiments may have different values. It should be noted that similar reference numerals and letters in the following figures denote similar items; therefore, once an item is defined in one figure, it need not be further discussed in subsequent figures.

[0045] In the description of this invention, it should be understood that the orientation or positional relationship indicated by directional terms such as "front, back, up, down, left, right", "horizontal, vertical, horizontal" and "top, bottom" is generally based on the orientation or positional relationship shown in the accompanying drawings, and is only for the convenience of describing this invention and simplifying the description. Unless otherwise stated, these directional terms 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 on the scope of protection of this invention; the directional terms "inner" and "outer" refer to the inner and outer contours relative to the outline of each component itself.

[0046] For ease of description, spatial relative terms such as "above," "on top of," "on the upper surface of," "above," etc., are used herein to describe the spatial positional relationship of a device or feature as shown in the figures to other devices or features. It should be understood that spatial relative terms are intended to encompass different orientations in use or operation beyond the orientation of the device as described in the figures. For example, if the device in the figures were inverted, a device described as "above" or "on top of" other devices or structures would subsequently be positioned as "below" or "under" other devices or structures. Thus, the exemplary term "above" can include both "above" and "below." The device may also be positioned in other different ways (rotated 90 degrees or in other orientations), and the spatial relative descriptions used herein will be interpreted accordingly.

[0047] Furthermore, it should be noted that the use of terms such as "first" and "second" to define components is merely for the purpose of distinguishing the corresponding components. Unless otherwise stated, the above terms have no special meaning and therefore should not be construed as limiting the scope of protection of this invention.

[0048] See Figures 1 to 17 As shown, according to an embodiment of the present invention, a POCT cartridge with an externally mounted nucleic acid extraction cavity is provided, comprising:

[0049] The reagent chamber body 1 has a plunger cavity 11 extending vertically through both ends, and a plurality of receiving cavities surrounding the plunger cavity 11. A plunger 3 is provided in the top opening of the plunger cavity 11. See details. Figure 6 As shown, each of the accommodating chambers includes a sample chamber 141, a waste liquid chamber 142, a proteinase K chamber 143, an elution liquid chamber 144, a first washing chamber 1451, a second washing chamber 1452, a third washing chamber 1453, a fourth washing chamber 1454, a fifth washing chamber 1455, and a lyophilized bulb chamber 145. Each accommodating chamber is relatively independent of the others. Each of the accommodating chambers has a top opening and a bottom wall. The overall exterior of the reagent chamber body 1 can be designed to be approximately cylindrical. The aforementioned plunger 3 can be driven by an external driving structure to reciprocate up and down, thereby realizing the transfer of liquid between the plunger chamber 11 and the various chambers.

[0050] Nucleic acid extraction cavity 2 is formed on the outer wall of reagent cavity body 1 and protrudes from the outer wall of reagent cavity body 1. The internal space of nucleic acid extraction cavity 2 can be controllably connected with plunger cavity 11. Nucleic acid extraction cavity 2 has a built-in magnetic bead (not shown in the figure). The bottom wall of nucleic acid extraction cavity 2 is constructed with a receiving groove (not shown in the figure), which is used to receive the ultrasonic head of the ultrasonic device.

[0051] In this technical solution, the nucleic acid extraction cavity 2 is set separately protruding from the outer wall of the reagent cavity body 1, instead of being arranged around the plunger cavity 11 with the aforementioned accommodating cavities. This allows for independent structural design of the nucleic acid extraction cavity 2, and a reliable match is formed between the accommodating groove on its bottom wall and the ultrasonic head. This enables efficient mixing of the sample liquid within the cavity using ultrasound, thereby improving the efficiency of the entire nucleic acid extraction and detection process. At the same time, since the nucleic acid extraction cavity 2 protrudes from the outer wall of the reagent cavity body 1, the distance between it and the accommodating cavities within the reagent cavity body 1 is increased, which effectively reduces the adverse interference of the ultrasonic device on the reagents or liquids in other adjacent accommodating cavities during operation. Furthermore, the protruding nucleic acid extraction cavity 2 can be more easily matched with the magnetic bead adsorption device, facilitating the effective adsorption of the magnetic beads within it.

[0052] See details Figure 13 As shown, the outer wall of the nucleic acid extraction chamber 2 is connected to the outer wall of the reagent chamber body 1 by a snap-fit ​​21. That is, a corresponding slot is provided on the outer wall of the reagent chamber body 1, and the snap-fit ​​21 matches the slot to achieve a reliable connection between the two. In this technical solution, the nucleic acid extraction chamber 2 can be detachably connected to the reagent chamber body 1 by a snap-fit ​​mechanism. This allows for the use of nucleic acid extraction chambers 2 of different specifications (e.g., capacity) before specific extraction and detection procedures, thereby meeting the needs of different nucleic acid extraction volumes. The nucleic acid extraction chamber 2 has a connecting pipe communicating with the inlet / outlet 22 of the first extraction chamber. This connecting pipe is inserted into the extraction chamber connection hole 1462 on the reagent chamber body 1, and a sealing ring 23 is used between the two to ensure a tight seal.

[0053] See details Figure 1 As shown, the external POCT cartridge of the nucleic acid extraction chamber also includes a flat reaction chamber 4, which is formed on the outer wall of the reagent chamber body 1 and protrudes from the outer wall of the reagent chamber body 1. The internal space of the flat reaction chamber 4 can be controllably connected to the plunger chamber 11. Furthermore, the outer wall of the flat reaction chamber 4 and the outer wall of the reagent chamber body 1 are detachably connected, for example, by an interference fit. It is understood that in order to ensure the assembly sealing between the aforementioned nucleic acid extraction chamber 2 and the flat reaction chamber 4 and the reagent chamber body 1, sealing rings 23 are embedded at the corresponding flow path interface positions of the two.

[0054] In this technical solution, the aforementioned flat reaction chamber 4 is detachably connected to the reagent chamber body 1. Different sizes of flat reaction chamber 4 can be selected and replaced according to different sample reaction amounts. In addition, it should be noted that the flat design of the reaction chamber can meet the requirements of large-area amplification and temperature control, while also facilitating optical detection at this location, thus achieving the purpose of real-time detection.

[0055] See also Figure 1 and Figure 10As shown, the external POCT cartridge for nucleic acid extraction chambers further includes: a rotary valve 5, comprising a valve body 51 and a protrusion 52 located on one side of the valve body 51. The protrusion 52 is inserted into the bottom opening of the plunger cavity 11. A flow channel (not shown in the figure) is constructed inside the rotary valve 5. The first port 511 of the flow channel is constructed on the top surface of the valve body 51, and the second port 512 of the flow channel is constructed on the free end face of the protrusion 52. The second port 512 extends along the axial direction of the protrusion 52. Each of the accommodating cavities has a chamber inlet / outlet port 12 constructed on its bottom wall. The bottom end face of the reagent chamber body 1 is also constructed with a surface that is connected to the flat... The first reaction chamber inlet / outlet 41, which is connected to the internal space of the flat reaction chamber 4, and the first extraction chamber inlet / outlet 22, which is connected to the internal space of the nucleic acid extraction chamber 2, can be aligned with one of the chamber inlet / outlet 12, the first reaction chamber inlet / outlet 41, and the first extraction chamber inlet / outlet 22 when the rotary valve 5 is driven to rotate. This allows the liquid flowing through the inlet / outlet aligned with the first port 511 to directly communicate with the plunger chamber 11 via the flow channel. For example, when the first port 511 is aligned with the first reaction chamber inlet / outlet 41, the plunger chamber 11 is connected to the internal space of the flat reaction chamber 4 only through the flow channel in the rotary valve 5.

[0056] In this technical solution, when the rotary valve 5 is driven to rotate, its flow channel can be aligned with one of the inlet / outlet ports 12 of each of the chambers, the inlet / outlet port 41 of the first reaction chamber, and the inlet / outlet port 22 of the first extraction chamber. This enables selective communication between each accommodating chamber, reaction chamber, or extraction chamber and the plunger chamber 11. Furthermore, the transfer of liquid can be achieved by controlling the plunger 3. More importantly, compared to the prior art method of guiding liquid flow by constructing flow channels corresponding to each chamber within the device base, the inlet / outlet ports of each chamber in this invention can directly transfer liquid via the flow channel on the rotary valve 5 without being guided by flow channels on the base. This effectively improves the utilization rate of sample solutions, reagents, and other liquids, effectively overcomes the shortcomings of the prior art where a large amount of liquid remains in the base, and thus reduces the cost of nucleic acid extraction and detection.

[0057] See details Figure 9 As shown, the inlet / outlet ports 12 of each chamber, the inlet / outlet port 41 of the first reaction chamber, and the inlet / outlet port 22 of the first extraction chamber are located on a first circle (not labeled in the figure). The first circle is coaxial with the protrusion 52, and the first port 511 is located on the first circle. In this way, the internal space of the plunger chamber 11 and each accommodating chamber, nucleic acid extraction chamber and flat reaction chamber can be controlled by simply rotating the rotary valve 5. The structure is simple and compact.

[0058] See Figure 11 As shown, a wedge-shaped groove 513 is constructed on the end face of the valve body 51 facing away from the reagent chamber body 1. In the specific application of this POCT cartridge, the rotary drive component in the external drive device can be fitted into the wedge-shaped groove 513. Since it is a wedge-shaped groove 513, it can achieve good drive positioning, thereby achieving stable and precise control of the rotary valve 5.

[0059] See details Figure 14 As shown, in some embodiments, the externally mounted POCT cartridge of the nucleic acid extraction chamber further includes: a base 6, which is detachably assembled to the bottom end of the reagent chamber body 1, and an assembly gap is formed between the base 6 and the bottom end of the reagent chamber body 1. The rotary valve 5 is rotatably connected within the assembly gap, and the bottom surface of the valve body 51 is flush with the bottom surface of the base 6. It should be noted that, as mentioned above, unlike the device base in the prior art, the base 6 in this invention does not have a corresponding liquid flow channel constructed inside; that is, the base 6 in this invention does not serve as a carrier for transferring liquid, but rather exists as an assembly and positioning structure for the rotary valve 5.

[0060] See further Figure 14 As shown, the base 6 includes a base body 61. The central area of ​​the top surface of the base body 61 has a connecting protrusion 62 extending toward the reagent chamber body 1. The connecting protrusion 62 is snapped together with the bottom end of the reagent chamber body 1, and the top surface of the connecting protrusion 62 is supported under the bottom end surface of the valve body 51.

[0061] Specifically, in this technical solution, the base 6 is inserted into the central blind hole at the bottom of the reagent chamber body 1 via a connecting protrusion 62. Multiple first locking protrusions 621 are spaced circumferentially around the outer periphery of the connecting protrusion 62 to engage with the wall of the central blind hole, thus achieving a reliable connection between the base 6 and the reagent chamber body 1. The bottom surface of the valve body 51 of the rotary valve 5 is reliably supported by the top surface of the connecting protrusion 62, preventing the rotary valve 5 from axially detaching from the reagent chamber body 1, ensuring structural compactness, and effectively preventing the potential for liquid leakage due to axial detachment of the rotary valve 5, thus ensuring the fully enclosed feature of the cartridge of this invention. In another specific embodiment, a sealing ring is fitted between the protrusion 52 and the inner wall of the plunger cavity 11 to further prevent liquid in the plunger cavity 11 from leaking out through the assembly gap between the two.

[0062] See you again Figure 14As shown, the top surface of the base body 61 is constructed with an anti-detachment structure (not labeled in the figure) extending toward the reagent chamber body 1. The anti-detachment structure includes spaced and oppositely arranged bent baffles 63. The amplification chamber 45 of the flat reaction chamber 4 extends out of the gap between the two bent baffles 63. The flat reaction chamber 4 has a connection and mating part 42 with the reagent chamber body 1. The connection and mating part 42 is clamped between the bent baffles 63 and the outer wall of the reagent chamber body 1.

[0063] In this technical solution, by setting an anti-detachment structure on the top surface of the base body 61, during the assembly process, the flat reaction chamber 4 is first inserted into the outer wall of the reagent chamber body 1 through the connecting fitting part 42. After the base 6 is assembled at the bottom end of the reagent chamber body 1, the anti-detachment structure covers the outside of the connecting fitting part 42 from bottom to top, thereby effectively preventing the flat reaction chamber 4 from detaching from the reagent chamber body 1. Setting the anti-detachment structure on the base 6 can facilitate the assembly of the flat reaction chamber 4 and the outer wall of the reagent chamber body 1.

[0064] See details Figures 15 to 17 As shown, the flat reaction chamber 4 has a second reaction chamber inlet / outlet 431 that can communicate with the plunger chamber 11 and a first outlet 432 that communicates with the ventilation channel in the reagent chamber body 1 (the ventilation channel extends from bottom to top along the axial direction of the reagent chamber body 1 to the vent hole 811 on the top cover 8 described later). The second reaction chamber inlet / outlet 431 and the first outlet 432 are both constructed as tubes and are respectively inserted into the reaction chamber connection hole 1461. The flat reaction chamber 4 has a main channel 441. One end of the main channel 441 is the second reaction chamber inlet / outlet 431, and the other end of the main channel 441 is connected to the first outlet 432. The main channel 441 has a branch channel 442. The end of the branch channel 442 is connected to the amplification chamber 45. The amplification chamber 45 has an amplification chamber vent 451. It is understood that the amplification chamber vent 451 is provided with a breathable but waterproof sealing membrane, such as a PTFE membrane.

[0065] In the prior art, the amplification chamber within the reaction chamber is directly connected to the ventilation channel of the reagent chamber body 1. Since the amplification chamber is pre-filled with the corresponding reaction reagents, when the sample liquid in the amplification chamber is full, any further addition of sample liquid will cause the excess sample liquid to continue flowing downstream along the ventilation channel originally used for venting. This results in the sample liquid carrying the pre-filled quantitative reagents out of the amplification chamber, leading to inaccurate amplification reaction and reduced detection accuracy. In contrast, in this application, the sample liquid enters through the main flow channel 441 and then through the branch flow channel 442. Inside the amplification chamber 45, because a breathable but water-impermeable sealing membrane is installed at the amplification chamber vent 451 of the amplification chamber 45, when the amplification chamber 45 is filled with sample liquid, the excess sample liquid will not enter the amplification chamber 45 under the pressure of the plunger 3. Instead, it will be connected to the first outlet 432 through the end outlet of the main channel 441 and finally enter the ventilation channel of the reagent chamber body 1 for storage. This effectively prevents the reduction of the amount of reaction reagent and also effectively reduces the amount of sample liquid remaining in the main channel 441, thereby ensuring the accuracy of subsequent amplification and optical detection results.

[0066] In some embodiments, the amplification chamber vent 451 is located at the highest point of the amplification chamber 45 to ensure that the sample solution is completely filled in the amplification chamber 45, that is, to ensure that the gas in the amplification chamber 45 is completely discharged.

[0067] See details Figure 15 As shown, the flat reaction chamber 4 has a first facade and a second facade (in the usage state). The first facade and the second facade are arranged opposite to each other. The amplification chamber vent 451 passes through the first facade and the second facade. A first membrane 461 is covered and connected to the first facade to seal the openings of the main channel 441, the branch channel 442, the amplification chamber 45, and the amplification chamber vent 451 on the first facade. A drain trough is constructed on the second facade. One end of the drain trough is connected to the outflow end of the main channel 441, and the other end of the drain trough is connected to the first outlet 432. A second membrane 462 is covered and connected to the second facade to seal the openings of the drain trough and the amplification chamber vent 451 on the second facade. The first membrane 461 is a PE membrane (capable of withstanding the high temperature during the amplification process), and the second membrane 462 is made of a breathable but waterproof material (e.g., a PTFE membrane). The second reaction chamber inlet / outlet 431 and the first outlet 432 are both constructed on the connecting and mating part 42.

[0068] In this technical solution, the sample liquid inlet channels and amplification chamber 45 are located on the first facade, while the ventilation structures are located on the second facade. This facilitates more convenient and efficient membrane coating on the corresponding sides. Specifically, a single complete PE membrane can be used on the first facade to ensure temperature control during sample liquid amplification, while a single complete PTFE membrane can be used on the second facade, allowing for air permeability but not water permeability, ensuring smooth gas exhaust from the chamber. This simplifies the manufacturing process. Specifically, the aforementioned first membrane 461 and second membrane 462 are blow-molded and can be connected to the reaction chamber using ultrasonic welding or thermal bonding.

[0069] See details Figure 17 As shown, with reference to the liquid flow direction in the main channel 441, the downstream of the main channel 441 has a resistance section 47, which is located between the outlet end of the main channel 441 and the branch channel 442.

[0070] In this technical solution, by setting a resistance section 47 on the main flow channel 441 downstream of each branch flow channel 442, it is ensured that the main flow channel 441 preferentially flows into the amplification chamber 45. After the sample liquid in the amplification chamber 45 is completely filled (i.e., filled with sample liquid), the flow resistance in the amplification chamber 45 is greater and higher than the resistance at the aforementioned resistance section 47. Excess sample liquid in the main flow channel 441 continues to flow downstream from the resistance section 47 and eventually flows into the downstream ventilation channel for storage, preventing excess sample liquid from remaining in the main flow channel 441 and affecting the detection results. See details. Figure 17 As shown, the resistance section 47 is a Tesla valve structure, and the liquid flow direction in the main flow channel 441 is the opposite of that of the Tesla valve structure. The structure is simple and does not require separate control.

[0071] The branch channel 442 and the main channel 441 form an angle α, where 15° ≤ α ≤ 45°. The end of the branch channel 442 furthest from the main channel 441 is closer to the outflow end of the main channel 441. Within this angle range, the sample solution in the main channel 441 can be more smoothly diverted into the branch channel 442 and enter the amplification chamber 45. Experiments have shown that α = 30° yields the best results.

[0072] The branch channels 442 are multiple, and the multiple branch channels 442 are arranged sequentially at intervals along the length direction of the main channel 441. Each branch channel 442 is provided with an amplification chamber 45 at its end, so that multiple different detection items can be performed simultaneously. The aforementioned branch channels 442 can all be located on one side of the main channel 441, such as the lower side. Of course, if the size of the flat reaction chamber 4 allows, the aforementioned branch channels 442 can also be distributed on the upper and lower sides of the main channel 441.

[0073] In some embodiments, the external POCT cartridge of the nucleic acid extraction chamber further includes: a first sealing gasket 7, which is circular and sandwiched between the top surface of the valve body 51 and the bottom surface of the reagent chamber body 1. The first sealing gasket 7 has liquid passage holes 71 corresponding to the liquid inlet / outlet 12 of each chamber, the liquid inlet / outlet 41 of the first reaction chamber, the liquid inlet / outlet 22 of the first extraction chamber, and the first port 511, and a through hole 72 for passing through the protrusion 52. By sandwiching the first sealing gasket 7 between the bottom surface of the valve body 51 and the reagent chamber body 1, liquid leakage from the rotating mating surface of the two can be prevented.

[0074] In one embodiment, a plurality of first positioning posts 131 and second positioning posts 132 are constructed on the bottom end face of the reagent chamber body 1. The first sealing gasket 7 has a first positioning through hole 731 corresponding to each of the first positioning posts 131 and a second positioning through hole 732 corresponding to each of the second positioning posts 132. Each of the first positioning posts 131 is evenly spaced along the outer circumference of the first sealing gasket 7, and each of the second positioning posts 132 is evenly spaced around the through hole 72 and located radially inside the first sealing gasket 7. In this way, by setting corresponding positioning structures on the outer circumference and inner circumference of the circular first sealing gasket 7, the frictional force applied to the first sealing gasket 7 by the rotary valve 5 during rotation can be effectively prevented from causing deformation of the first sealing gasket 7, thereby reducing the risk of leakage at this point. In another embodiment, the bottom end face of the reagent chamber body 1 and the first sealing gasket 7 are integrally injection molded. By integrally injection molding, leakage caused by misalignment and deformation of the first sealing gasket 7 during the rotation of the rotary valve 5 can also be effectively prevented.

[0075] The top of the reagent chamber body 1 is fastened with an upper cover 8, which seals the top openings of each accommodating chamber (including the ventilation channel mentioned above). The upper cover 8 includes a cover body 81 and a cap 82 that are fastened together. The cover body 81 has ventilation holes 811 corresponding to each accommodating chamber to facilitate the discharge of gas during the pushing and pulling of the plunger 3 and ensure smooth liquid transfer. The cover body 81 and the cap 82 are respectively provided with concentric plunger passage holes 814. The cover body 81 also has a sample loading hole 812. The cover body 81 is fastened to the reagent chamber body 1 by a second latching protrusion 813. A second sealing gasket 9 is sandwiched between the top end face of the upper cover 8 and the reagent chamber body 1. The second sealing gasket 9 has circular holes corresponding one-to-one with the vent holes 811 on the cover 81. The size of the circular holes is proportional to the size of the vent holes 811, preferably 0.8-1.2 times the size of the corresponding vent holes 811. The second sealing gasket 9 also has an opening corresponding to the position of the sample application hole 812. This opening can be configured as a fan or a quadrilateral, with a size equal to the circumscribed quadrilateral or fan shape of the sample application hole 812. This design ensures both the sealing of the cartridge and user convenience. The second sealing gasket 9 also has an opening corresponding to the plunger through-hole 814. The cap 82 is covered with a breathable but waterproof sealing membrane. The aforementioned first and second sealing gaskets can specifically be silicone gaskets.

[0076] Each of the individual components mentioned above is manufactured using injection molding.

[0077] The following describes the specific application process of the card holder of the present invention:

[0078] The POCT cartridge externally mounted on the nucleic acid extraction chamber of the present invention is used in conjunction with the corresponding nucleic acid detection equipment (hereinafter referred to as the equipment). The equipment is equipped with a plunger rod, which pushes the plunger to create a pressure difference between the chamber of the main reagent chamber (i.e., the main reagent chamber 1 mentioned above, the same below) and the plunger chamber of the rotary valve (i.e., the plunger chamber 11 mentioned above, the same below). The rotation of the equipment motor drives the cartridge rotary valve (i.e., the aforementioned rotary valve 5, the same below) to rotate. The rotary valve is equipped with a liquid path (i.e., the aforementioned flow channel). One end of the liquid path of the rotary valve is connected to the flow channel ports distributed in a circular pattern on the main reagent chamber (i.e., the chamber inlet / outlet 12, the first reaction chamber inlet / outlet 41, and the first extraction chamber inlet / outlet 22 mentioned above) by the rotation angle of the motor. The other end is connected to the internal space of the plunger chamber, thereby realizing the transfer of liquid from one chamber to another through the plunger chamber of the rotary valve.

[0079] (1) After the user completes the sampling, add the sample to be tested through the sample inlet (i.e., the sample inlet 812 mentioned above) of the main reagent chamber, put on the cap (i.e., the cap 82 mentioned above), and put it into the device.

[0080] (2) During use, the rotating component of the device connects one end of the rotary valve to the flow channel of the sample chamber 141, pulling the plunger 3 to generate a pressure difference, causing the liquid in the sample chamber 141 to enter the rotary valve plunger chamber. The motor rotates, causing the rotary valve to rotate to a specific angle, and then pushes the plunger rod to transfer the reagent inside the rotary valve plunger chamber to the proteinase K chamber for mixing with proteinase K. Pulling the plunger 3 again generates a pressure difference, causing the liquid in the proteinase K chamber to enter the rotary valve plunger chamber, and then pushes the plunger rod to transfer the reagent inside the rotary valve plunger chamber to the extraction chamber. Magnetic beads are pre-set in the extraction chamber. The ultrasonic device is energized to mix the magnetic beads, enabling the magnetic beads to effectively adsorb nucleic acids. The instrument's magnetic bead adsorption device adsorbs the magnetic beads. The motor rotates, causing the rotary valve to rotate to a specific angle, pulling the plunger 3 to generate a pressure difference, drawing the liquid into the rotary valve plunger chamber. The motor rotates, causing the rotary valve to rotate to a specific angle, pushing the plunger 3 to generate a pressure difference, and then transferring the liquid to the waste liquid chamber 142.

[0081] (3) The motor rotates and drives the rotary valve to rotate to a specific angle, pulling the plunger 3 to generate a pressure difference, transferring the liquid in the first washing chamber 1451 into the rotary valve plunger chamber. The motor rotates and drives the rotary valve to rotate to a specific angle, pushing the plunger 3 to generate a pressure difference, transferring the liquid to the extraction chamber (that is, the internal space of the nucleic acid extraction chamber 2 mentioned above, the same below). The ultrasonic device is powered on to mix the magnetic beads. After washing, the magnetic bead adsorption device of the instrument adsorbs the magnetic beads. The motor rotates and drives the rotary valve to rotate to a specific angle, pulling the plunger 3 to generate a pressure difference, drawing the liquid into the plunger chamber. The motor rotates and drives the rotary valve to rotate to a specific angle, pushing the plunger to generate a pressure difference, and then transferring the liquid to the waste liquid chamber 142.

[0082] (4) The motor rotates and drives the rotary valve to rotate to a specific angle, pulling the plunger 3 to generate a pressure difference, transferring the liquid in the second washing chamber 1452 into the plunger chamber. The motor rotates and drives the rotary valve to rotate to a specific angle, pushing the plunger to generate a pressure difference, transferring the liquid to the extraction chamber. The ultrasonic device is powered on to mix the magnetic beads. After washing, the magnetic bead adsorption device of the instrument adsorbs the magnetic beads. The motor rotates and drives the rotary valve to rotate to a specific angle, pulling the plunger to generate a pressure difference, drawing the liquid into the plunger chamber. The motor rotates and drives the rotary valve to rotate to a specific angle, pushing the plunger to generate a pressure difference, and then transferring the liquid to the waste liquid chamber 142.

[0083] (5) The motor rotates and drives the rotary valve to rotate to a specific angle, pulling the plunger to generate a pressure difference, transferring the liquid in the eluent chamber 144 into the plunger chamber. The motor rotates and drives the rotary valve to rotate to a specific angle, pushing the plunger to generate a pressure difference, transferring the liquid into the extraction chamber. The ultrasonic device is powered on to mix the magnetic beads. After washing, the magnetic bead adsorption device of the instrument adsorbs the magnetic beads. The motor rotates and drives the rotary valve to rotate to a specific angle, pulling the plunger to generate a pressure difference, drawing the liquid into the plunger chamber. The motor rotates and drives the rotary valve to rotate to a specific angle, pushing the plunger to generate a pressure difference, and then transferring the liquid into the freeze-dried bulb chamber 145 to complete the freeze-dried bulb reconstitution.

[0084] (6) The motor rotates and drives the rotary valve to rotate to a specific angle, which pulls the plunger to generate a pressure difference, and transfers the liquid in the elution chamber into the plunger chamber. The motor rotates and drives the rotary valve to rotate to a specific angle, which pushes the plunger to generate a pressure difference, and transfers the liquid into the reaction chamber (that is, the internal space of the flat reaction chamber 4 mentioned above).

[0085] It should be noted that the third to fifth washing chambers can be selected independently to match different testing items.

[0086] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention. The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the technical principles of the present invention, and these improvements and modifications should also be considered within the protection scope of the present invention.

Claims

1. A POCT cartridge with an externally mounted nucleic acid extraction chamber, characterized in that, include: The reagent chamber body (1) has a plunger cavity (11) that extends through both ends of the reagent chamber body (1) and a plurality of accommodating cavities arranged around the plunger cavity (11). A plunger (3) is provided in the top opening of the plunger cavity (11). Nucleic acid extraction cavity (2) is formed on the outer wall of the reagent cavity body (1) and protrudes from the outer wall of the reagent cavity body (1). The internal space of the nucleic acid extraction cavity (2) can be controllably connected with the plunger cavity (11). The nucleic acid extraction cavity (2) is equipped with magnetic beads. The bottom wall of the nucleic acid extraction cavity (2) is constructed with a receiving groove for receiving the ultrasonic head of the ultrasonic device. It also includes a flat reaction chamber (4), which is formed on the outer wall of the reagent chamber body (1) and protrudes from the outer wall of the reagent chamber body (1). The internal space of the flat reaction chamber (4) can be controllably connected with the plunger chamber (11). Also includes: A rotary valve (5) includes a valve body (51) and a protrusion (52) on one side of the valve body (51). The protrusion (52) is inserted into the bottom opening of the plunger cavity (11). A flow channel is constructed inside the rotary valve (5). The first port (511) of the flow channel is constructed on the top surface of the valve body (51). The second port (512) of the flow channel is constructed on the free end face of the protrusion (52), and the second port (512) extends along the axial direction of the protrusion (52). Each of the accommodating cavities has a chamber inlet / outlet port (12) on its bottom wall. The reagent chamber... The bottom end face of the main body (1) is also constructed with a first reaction chamber inlet / outlet (41) communicating with the internal space of the flat reaction chamber (4) and a first extraction chamber inlet / outlet (22) communicating with the internal space of the nucleic acid extraction chamber (2). When the rotary valve (5) is driven to rotate, the first port (511) can be aligned with one of the chamber inlet / outlet (12), the first reaction chamber inlet / outlet (41), and the first extraction chamber inlet / outlet (22) so that the liquid flowing through the inlet / outlet aligned with the first port (511) can be directly connected to the plunger cavity (11) through the flow channel.

2. The POCT card holder according to claim 1, characterized in that, The outer wall of the nucleic acid extraction cavity (2) is fastened to the outer wall of the reagent cavity body (1) by a snap fastener (21).

3. The POCT card holder according to claim 2, characterized in that, Each of the chamber inlet / outlet (12), the first reaction chamber inlet / outlet (41), and the first extraction chamber inlet / outlet (22) is located on a first circle, the first circle is coaxial with the protrusion (52), and the first port (511) is located on the first circle; and / or, the valve body (51) has a wedge-shaped groove (513) on the end face away from the reagent chamber body (1).

4. The POCT card holder according to claim 3, characterized in that, Also includes: The base (6) is detachably assembled to the bottom end of the reagent chamber body (1), and an assembly gap is formed between the base (6) and the bottom end of the reagent chamber body (1). The rotary valve (5) is rotatably connected to the assembly gap, and the bottom surface of the valve body (51) is flush with the bottom surface of the base (6).

5. The POCT card holder according to claim 4, characterized in that, The base (6) includes a base body (61), and the central area of ​​the top surface of the base body (61) has a connecting protrusion (62) extending toward the reagent chamber body (1). The connecting protrusion (62) is snapped together with the bottom end of the reagent chamber body (1), and the top surface of the connecting protrusion (62) is supported under the bottom end surface of the valve body (51).

6. The POCT card holder according to claim 5, characterized in that, Also includes: The first sealing gasket (7) is circular and is sandwiched between the top surface of the valve body (51) and the bottom surface of the reagent chamber body (1). The outer wall of the flat reaction chamber (4) is detachably connected to the outer wall of the reagent chamber body (1).

7. The POCT card holder according to claim 6, characterized in that, The base body (61) has an anti-detachment structure extending toward the reagent chamber body (1) on its top surface. The anti-detachment structure includes spaced and oppositely arranged bent baffles (63). The amplification chamber (45) of the flat reaction chamber (4) extends out of the gap between the two bent baffles (63). The flat reaction chamber (4) has a connection and mating part (42) with the reagent chamber body (1). The connection and mating part (42) is clamped between the bent baffle (63) and the outer wall of the reagent chamber body (1).

8. The POCT card holder according to claim 6, characterized in that, Also includes: The first sealing gasket (7) has a liquid passage hole (71) that corresponds to the liquid inlet / outlet (12) of each of the chambers, the liquid inlet / outlet (41) of the first reaction chamber, the liquid inlet / outlet (22) of the first extraction chamber, and the first port (511), and a perforation (72) for passing through the protrusion (52).

9. The POCT card holder according to claim 8, characterized in that, The bottom end face of the reagent chamber body (1) is provided with a plurality of first positioning posts (131) and second positioning posts (132). The first sealing gasket (7) is provided with a first positioning through hole (731) corresponding to each of the first positioning posts (131) and a second positioning through hole (732) corresponding to each of the second positioning posts (132). Each of the first positioning posts (131) is evenly spaced along the outer circumference of the first sealing gasket (7), and each of the second positioning posts (132) is evenly spaced around the through hole (72) and is located on the radial inner side of the first sealing gasket (7); and / or, the bottom end face of the reagent chamber body (1) is integrally injection molded with the first sealing gasket (7).

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