A multiplex primer kit for enrichment of a target region of a gene

Abstract

This invention provides a multiplex primer kit for enriching gene target regions, relating to the fields of precision medicine and molecular diagnostics. The kit includes a placement tray on which catalyst tubes, detergent tubes, standard tubes, and a microplate are distributed. The invention utilizes a self-sealing gasket within the sealing cap to provide the first seal for the microplate's wells. Simultaneously, the sealing plate mechanism of the pressing mechanism provides a secondary seal by pressing the sealing cap with a sealing plug. Combined with the locking mechanism of the box body, this forms a complete sealing system from the wells to the box body, effectively blocking external moisture and contaminants, significantly extending the shelf life of primers and reagents. The locking mechanism allows for the dedicated clamping and fixing of reagent tubes such as catalyst and detergent tubes. Limiting protrusions limit the position of the microplate. Combined with the pressing action of the first and second pressing blocks of the pressing plate mechanism, the problem of reagent tube displacement and collision during transportation is completely solved, preventing container damage and reagent leakage.

Description

Technical Field

[0001] This invention relates to the fields of precision medicine and molecular diagnostics, and in particular to a multiplex primer kit for enriching gene target regions. Background Technology

[0002] Gene target region enrichment technology uses multiple primers to specifically capture and amplify target gene fragments, enabling efficient detection of disease-related gene loci. In the clinical and research applications of this technology, multiple primers, accompanying catalysts, detergents, and other reagents must be stored and transported under low-temperature, dry, and sealed conditions; otherwise, decreased primer activity and reagent contamination will directly lead to false negatives or false positives in the test results.

[0003] In the prior art, such as the "Primer Set and Kit Based on Multiplex PCR" patent application number CN202110578727.8, the primer set provided by this invention can simultaneously perform multiplex PCR capture sequencing of 15 genes, detecting a total of 93 different mutations. The primers have high specificity, with a depth of over 1000X for tissue samples and over 10000X for blood samples, with an average coverage of 99%. The template starting amount is small, which can efficiently, rapidly and conveniently detect mutations in lung cancer, colorectal cancer, pancreatic cancer and gastrointestinal stromal tumors, thus developing a highly sensitive detection method with broad application prospects.

[0004] Traditional reagent kits rely on simple adhesion between the box and the lid, and the wells of microplates are sealed with ordinary lids, which cannot effectively isolate external moisture and microorganisms, resulting in reduced primer activity, reagent contamination, and affecting the stability of test results. Summary of the Invention

[0005] The purpose of this invention is to solve the problems in the prior art where the box body and lid are simply attached together, and the wells of the microplate are sealed only by ordinary lids, which cannot effectively isolate external moisture and microorganisms, resulting in reduced primer activity, reagent contamination, and affecting the stability of detection results. Therefore, this invention proposes a multiplex primer kit for enriching gene target regions.

[0006] To achieve the above objectives, the present invention adopts the following technical solution: a multiplex primer kit for enriching gene target regions, comprising a kit, wherein a locking mechanism is provided on the side of the kit, and a placement mechanism and a pressing mechanism are provided on the inner side of the kit, wherein the pressing mechanism is disposed above the placement mechanism, and the kit comprises a lid and a base. The placement mechanism includes a placement tray on which catalyst tubes, detergent tubes, standard tubes and microporous plates are distributed. Several plate holes are evenly opened through the microporous plates. A sealing cap is provided on the top of each plate hole, and a self-sealing gasket is provided on the inner side of the sealing cap for sealing. The pressure plate mechanism includes a sealing pressure plate, on which several sealing plugs are evenly distributed. Each sealing plug is pressed against the top of the sealing cover for sealing.

[0007] Preferably, the locking mechanism includes a side mount, on the inner side of which a toggle handle is rotatably mounted, and the side mount is fixedly mounted on the side of the lid.

[0008] Preferably, a bidirectional adjusting screw is connected through the inner side of the toggle handle, and a transmission component is symmetrically threaded onto the outer wall of the bidirectional adjusting screw.

[0009] Preferably, the transmission component is provided with a limiting sleeve on its side, and the box cover and box base are symmetrically fixedly installed with limiting fasteners on their front sides, with the limiting sleeve slidably connected to the outside of the limiting fastener.

[0010] Preferably, a number of clips are evenly distributed on the top of the placement tray, and the clips are symmetrically attached to the outer side of the catalyst tube.

[0011] Preferably, the top of the placement tray has a placement groove, and the inner two ends of the placement groove are symmetrically fixedly installed with limiting protrusions, which are located at both ends of the microporous plate.

[0012] Preferably, the pressing plate mechanism further includes a pressing plate, on which a plurality of first pressing blocks and second pressing blocks are evenly distributed, and the pressing plate is provided with pressing grooves.

[0013] Preferably, a hinge is movably mounted on the side of the lid, and the hinge is connected to the side of the box base.

[0014] Compared with the prior art, the advantages and positive effects of the present invention are as follows: 1. In this invention, the reagent kit uses a self-sealing gasket built into the sealing cap to provide the first seal for the wells of the microplate. Simultaneously, the sealing plate of the pressure plate mechanism provides a second seal by pressing the sealing cap with a sealing plug. Combined with the locking mechanism of the box body, a complete sealing system from the wells to the box body is formed, effectively blocking external moisture and contaminants, significantly extending the shelf life of primers and reagents. The clips in the placement mechanism can be used to secure reagent tubes such as catalyst tubes and detergent tubes. The limiting protrusions limit the microplate, and together with the first and second pressing blocks of the pressure plate mechanism, the pressing action of the reagent tubes completely solves the problem of reagent tube displacement and collision during transportation, avoiding container damage and reagent leakage.

[0015] 2. In this invention, the locking mechanism drives the bidirectional adjusting screw to rotate by turning the handle, so that the transmission components on both sides drive the limiting sleeve to slide synchronously along the limiting fastener, thereby achieving uniform locking of the box lid and the box seat. Compared with traditional buckles, the locking force is greater and the force is more even, effectively avoiding the risk of the box lid opening accidentally during low temperature transportation or frequent handling. Attached Figure Description

[0016] Figure 1 A schematic diagram of the three-dimensional structure of a multiplex primer kit for enriching gene target regions proposed in this invention; Figure 2 This is a schematic diagram of another aspect of the structure of a multiplex primer kit for enriching gene target regions proposed in this invention. Figure 3 The diagram shows the unfolded structure of a multiplex primer kit for enriching gene target regions proposed in this invention. Figure 4 For the present invention Figure 3 Enlarged structural diagram at point A in the diagram; Figure 5 This invention presents a schematic diagram of the placement mechanism of a multiplex primer kit for enriching gene target regions. Figure 6 A schematic diagram of the pressure plate mechanism of a multiplex primer kit for enriching gene target regions proposed in this invention; Figure 7 For the present invention Figure 6 A magnified structural diagram at point B in the diagram.

[0017] Legend: 1. Reagent kit; 101. Box lid; 102. Box base; 103. Hinge; 2. Locking mechanism; 201. Side mounting base; 202. Actuating handle; 203. Bidirectional adjusting screw; 204. Transmission component; 205. Limiting sleeve; 206. Limiting fastener; 3. Placement mechanism; 301. Placement tray; 302. Catalyst tube; 303. Clamp; 305. Detergent tube; 306. Standard tube; 307. Placement slot; 308. Microplate; 309. Plate hole; 310. Sealing cap; 311. Self-sealing gasket; 312. Limiting protrusion; 4. Pressing plate mechanism; 401. Pressing plate; 402. First pressing block; 403. Second pressing block; 404. Pressing groove; 405. Sealing pressing plate; 406. Sealing plug. Detailed Implementation

[0018] To better understand the above-mentioned objectives, features, and advantages of the present invention, the present invention will be further described below in conjunction with the accompanying drawings and embodiments. It should be noted that, unless otherwise specified, the embodiments and features described in these embodiments can be combined with each other.

[0019] Numerous specific details are set forth in the following description in order to provide a full understanding of the invention. However, the invention may also be practiced in other ways than those described herein, and therefore the invention is not limited to the specific embodiments disclosed in the following specification.

[0020] Example 1: As Figures 1-7 As shown, the present invention provides a technical solution: a multiplex primer kit for enriching gene target regions, comprising a kit 1, a locking mechanism 2 provided on the side of the kit 1, a placement mechanism 3 and a pressing mechanism 4 provided on the inner side of the kit 1, the pressing mechanism 4 being disposed above the placement mechanism 3, the kit 1 comprising a lid 101 and a base 102, a hinge 103 being movably mounted on the side of the lid 101, the hinge 103 being connected to the side of the base 102; The placement mechanism 3 includes a placement tray 301, on which are distributed catalyst tubes 302, detergent tubes 305, standard tubes 306 and microporous plates 308. A plurality of plate holes 309 are evenly distributed through the microporous plates 308. A sealing cap 310 is provided on the top of the plate holes 309. A self-sealing gasket 311 is provided on the inner side of the sealing cap 310 for sealing. A plurality of clips 303 are evenly distributed on the top of the placement tray 301. The clips 303 are symmetrically clipped onto the outer side of the catalyst tubes 302. A placement groove 307 is provided on the top of the placement tray 301. Limiting protrusions 312 are symmetrically fixedly installed at both ends of the inner side of the placement groove 307. The limiting protrusions 312 are located at both ends of the microporous plates 308. The pressure plate mechanism 4 includes a sealing pressure plate 405, on which several sealing plugs 406 are evenly distributed. The sealing plugs 406 are all pressed against the top of the sealing cover 310 for pressing and sealing. The pressure plate mechanism 4 also includes a pressing plate 401, on which several first pressing blocks 402 and second pressing blocks 403 are evenly distributed. The pressing plate 401 is provided with a pressing groove 404.

[0021] In this embodiment, the box body is assembled from a lid 101, a base 102, and a hinge 103. As the basic structural component of the multiplex primer kit for enriching the entire gene target region, it provides a stable overall support foundation and a comprehensive external protective frame for various functional mechanisms, reagent tubes, and microplates inside the kit, effectively isolating external physical collisions and environmental interference. The hinge 103 connects the side of the lid 101 and the base 102, allowing the lid 101 to be flexibly flipped open and closed relative to the base 102, ensuring the smoothness and flexibility of daily opening and closing of the lid. At the same time, it can cooperate with the locking mechanism on the outside of the kit. After the lid 101 is fastened to the base 102, the locking mechanism achieves a tight closure of the entire box body, ensuring the structural stability and sealing reliability of the closed box body.

[0022] The placement tray 301, as the core supporting component of the placement mechanism, scientifically integrates functional areas such as a reagent tube clamping area and a microplate placement slot 307. Each area is rationally arranged according to the usage requirements of the reagent kit, enabling the orderly placement of different types of reagent tubes and microplates. The clamps 303 are evenly distributed along the reagent tube clamping area, ensuring precise clamping and positioning of various reagent tubes such as catalyst tubes 302, detergent tubes 305, and standard tubes 306, preventing shaking during storage or transportation. The displacement and limiting protrusions 312 are symmetrically arranged at both ends of the inner side of the microplate placement slot 307, which can accurately limit and fix the microplate 308 placed in the slot, preventing the microplate from shifting or sliding. The sealing cap 310, which is adapted to be installed on the top of the microplate hole 309, has a self-sealing gasket 311 embedded in its inner side, which can fit tightly with the port of the hole 309, to achieve primary sealing protection of the microplate hole, providing a basic and stable storage environment for various primers, reagents and supporting reagents in the kit. Meanwhile, the self-sealing gasket 311 built into the sealing cap 310 can form a tight seal with the port of the plate hole 309 of the microplate 308, providing the first layer of sealing protection for the primer reagents inside the plate hole, effectively preventing external impurities from directly contacting the reagents inside the plate hole; in conjunction with this, the sealing plugs 406 evenly distributed on the sealing pressure plate 405 in the pressure plate mechanism 4 can precisely press against the top of each sealing cap 310 when the box is closed, forming a uniform and stable pressing force on the sealing cap 310, further enhancing the sealing effect between the sealing cap 310 and the plate hole 309, and achieving a secondary pressing seal for the plate hole; this primary The sealing structure, which combines a sealed base with a secondary compression seal, along with the locking mechanism 2 on the outside of the box to lock the lid 101 and the base 102 together, forms a layered and synergistic sealing system that ultimately creates a complete chain of all-round sealing protection from the plate wells 309 to the microplate 308 and then to the entire kit box. This effectively prevents external moisture, dust, microorganisms, and other contaminants from entering the kit and contacting the reagents, significantly reducing the probability of primers and various supporting reagents becoming inactive or contaminated due to environmental factors, significantly extending the shelf life of primers and reagents, and ensuring the accuracy and reliability of subsequent gene target region enrichment detection experiments.

[0023] The pressing plate 401 in the pressing mechanism 4 serves as the core supporting foundation. Its surface is precisely equipped with a first pressing block 402 and a second pressing block 403 according to the arrangement of the reagent tubes. When the box is closed, the first pressing block 402 and the second pressing block 403 can precisely fit against the tops of various reagent tubes on the placement mechanism 3, forming a vertically downward pressing force on the reagent tubes. This achieves efficient pressing and limiting of the reagent tubes, preventing them from moving up and down. Simultaneously, the pressing plate 401 is equipped with a sealing pressing plate 405, which is divided according to the holes 309 of the microporous plate 308. The container has a number of sealing plugs 406 evenly arranged in a regular pattern. When the container is closed, each sealing plug 406 can be precisely aligned with the sealing cap 310 on the top of the microplate 308 well 309, and form a stable downward pressure on the sealing cap 310. The sealing plugs 406 further enhance the sealing fit between the sealing cap 310 and the well 309 through the pressing action of the sealing plugs 406, and achieve secondary pressing and sealing of the well 309. From the perspective of sealing protection, this ensures the stability of various primers and supporting reagents in the reagent kit during the entire process of storage and transportation, and reduces the probability of the reagents being affected by the external environment. The placement tray 301 of the placement mechanism 3 is equipped with dedicated clips 303 for the placement positions of reagent tubes of different specifications, such as catalyst tubes 302, detergent tubes 305, and standard tubes 306. The inner dimensions of each clip 303 are adapted to the outer wall dimensions of the corresponding reagent tube, which can firmly fix various reagent tubes and limit the displacement of the reagent tubes in the horizontal direction. At the same time, limiting protrusions 312 are symmetrically fixed at both ends of the inner side of the microporous plate placement groove 307. After the microporous plate 308 is placed in the placement groove 307, the limiting protrusions 312 can precisely limit the microporous plate 308 from both ends, effectively preventing the microporous plate 308 from shifting or sliding in the horizontal direction during storage and transportation. The first pressing block 402 and the second pressing block 403 on the pressing plate 401 of the pressing plate mechanism 4 can precisely abut against the top of the reagent tube after the box is closed, forming a stable pressing limit on the reagent tube from the vertical direction. The horizontal clamping and the vertical pressing limit form a double fixing structure that works together to restrict the displacement space of the reagent tube in all directions. This completely solves the problems of reagent tube displacement and collision caused by bumps and shaking during the transportation of the reagent kit, effectively avoids container damage caused by collision of the reagent tube, and eliminates reagent leakage caused by container damage, ensuring the safety and integrity of the reagents stored in the reagent kit.

[0024] Example 2: As Figures 1-7As shown, the locking mechanism 2 includes a side mounting base 201. A toggle handle 202 is rotatably mounted on the inner side of the side mounting base 201. The side mounting base 201 is fixedly mounted on the side of the cover 101. A bidirectional adjusting screw 203 is connected through the inner side of the toggle handle 202. A transmission component 204 is symmetrically threaded on the outer wall of the bidirectional adjusting screw 203. A limiting sleeve 205 is provided on the side of the transmission component 204. Limiting fasteners 206 are symmetrically fixedly mounted on the front of the cover 101 and the base 102. The limiting sleeve 205 is slidably connected to the outer side of the limiting fastener 206.

[0025] In this embodiment, the side mounting base 201 serves as the core mounting base of the locking mechanism 2, and is securely fixed to the side of the box cover 101 of the box body 1. Its interior provides suitable installation and movement space for adjusting components such as the bidirectional adjusting screw 203 and the transmission component 204, providing a stable support foundation for each adjusting component and ensuring the smoothness of the entire locking mechanism's adjustment action and structural stability. The bidirectional adjusting screw 203 is rotatably mounted inside the side mounting base 201, and its two ends are provided with external threads of opposite directions, enabling precise thread engagement with the transmission components 204 on both sides. When the adjusting screw 203 is rotated, the rotational motion is smoothly converted into the linear opposing or disengaging motion of the two transmission components 204 through the force of the threaded transmission. The transmission component 204 and the limiting sleeve 205 are an integrated fixed structure, which can synchronously drive the limiting sleeve 205 to slide precisely in a straight line along the limiting fastener 206 on the box body. Finally, the box cover 101 and the box base 102 are tightly closed by the locking action of the limiting sleeve 205 and the limiting fastener 206. The structural design completely solves the technical problem of easy loosening and disengagement of the traditional reagent kit snap-locking structure.

[0026] The locking mechanism 2 is manually adjustable. The lever 202 is coaxially fixed to the end of the bidirectional adjusting screw 203, serving as the power input component for the entire locking mechanism. The operator can easily drive the bidirectional adjusting screw 203 to rotate synchronously by turning the lever 202. During rotation, the bidirectional adjusting screw 203, through its oppositely oriented threads at both ends, simultaneously drives the transmission components 204 on both sides to slide synchronously and in the same direction horizontally. The transmission components 204 then synchronously drive the connected limiting sleeves 205 to slide precisely and synchronously along the limiting fasteners 206 on the cover 101 and the base 102, ensuring that the limiting sleeves 205 on both sides are evenly engaged in the preset locking positions of the limiting fasteners 206, thereby achieving… The lid 101 and the base 102 are locked in a uniform and omnidirectional manner. Compared with the simple snap-locking structure of traditional reagent kits, this screw-driven locking method can provide a locking force that is much greater than that of traditional snaps. Moreover, the locking force can be evenly distributed on the entire closed contact surface of the lid 101 and the base 102 through the screw drive, making the force on the box body more balanced. This effectively avoids the problem of box body deformation caused by excessive local force. Even if the plastic parts are slightly deformed in the environment of low temperature cold chain transportation, or if the reagent kit is subjected to bumps and shaking during frequent handling and movement in the laboratory, it can always maintain a stable locking state. This fundamentally avoids the risk of accidental opening of the lid 101 and ensures the safety of the reagents stored inside the reagent kit.

[0027] The working principle of this embodiment is as follows: When using the reagent kit, firstly, the lever 202 of the locking mechanism 2 is turned. The lever 202 drives the bidirectional adjusting screw 203 to rotate. The bidirectional adjusting screw 203 drives the transmission components 204 on both sides to move away from each other through threaded transmission. The transmission components 204 drive the limiting sleeve 205 to slide outward along the limiting fastener 206, releasing the locking of the lid 101 and the base 102. Then, the lid 101 is opened by flipping it upward through the hinge 103. The lid 101 drives the pressure plate mechanism 4 to lift synchronously. At this time, the sealing plug 406 of the sealing pressure plate 405 is disengaged from the sealing cover 310, and the first pressure block 402 and the second pressure block 403 are released from pressing the reagent tube. Then, the catalyst can be directly removed from the placement tray 301. Reagent tubes such as tube 302 and detergent tube 305 can be used, or primer reagents can be taken out from the plate hole 309 of microplate 308 by opening the sealing cover 310. After use, the reagent tube clips are put back into the clip 303, the microplate 308 is placed in the placement groove 307 and limited by the limiting protrusion 312, the sealing cover 310 is closed so that the self-sealing gasket 311 seals the plate hole 309, and then the box cover 101 is flipped down so that the sealing plug 406 of the pressure plate mechanism 4 is pressed against the top of the sealing cover 310, and the first pressure block 402 and the second pressure block 403 are pressed against the top of the reagent tube. Finally, the lever 202 is turned in the opposite direction, and the bidirectional adjusting screw 203 drives the transmission component 204 to move closer to each other, so that the limiting sleeve 205 slides inward along the limiting fastener 206 and locks the box body, completing the sealed storage.

[0028] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any other way. Any person skilled in the art may make changes or modifications to the above-disclosed technical content to create equivalent embodiments for application in other fields. However, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the protection scope of the present invention.

Claims

1. A multiplex primer kit for enriching gene target regions, comprising a kit (1), characterized in that: The reagent kit (1) is provided with a locking mechanism (2) on its side, and a placement mechanism (3) and a pressing mechanism (4) are provided on the inner side of the reagent kit (1). The pressing mechanism (4) is located above the placement mechanism (3). The reagent kit (1) includes a box cover (101) and a box base (102). The placement mechanism (3) includes a placement tray (301), on which are distributed catalyst tubes (302), detergent tubes (305), standard tubes (306) and microporous plates (308). A plurality of plate holes (309) are uniformly opened through the microporous plate (308), and a sealing cap (310) is provided on the top of the plate hole (309). A self-sealing gasket (311) is provided on the inner side of the sealing cap (310) for sealing. The pressure plate mechanism (4) includes a sealing pressure plate (405), on which a plurality of sealing plugs (406) are evenly distributed. The sealing plugs (406) are all pressed against the top of the sealing cover (310) for pressing and sealing.

2. The multiplex primer kit for enriching gene target regions according to claim 1, characterized in that: The locking mechanism (2) includes a side mount (201), on which a toggle handle (202) is rotatably mounted, and the side mount (201) is fixedly mounted on the side of the box cover (101).

3. The multiplex primer kit for enriching gene target regions according to claim 2, characterized in that: The inner side of the toggle handle (202) is connected to a bidirectional adjusting screw (203), and the outer wall of the bidirectional adjusting screw (203) is symmetrically threaded with a transmission component (204).

4. The multiplex primer kit for enriching gene target regions according to claim 3, characterized in that: The transmission component (204) is provided with a limiting sleeve (205) on its side, and the box cover (101) and the box base (102) are symmetrically fixedly installed with limiting fasteners (206) on their front sides. The limiting sleeve (205) is slidably connected to the outside of the limiting fastener (206).

5. The multiplex primer kit for enriching gene target regions according to claim 1, characterized in that: The top of the placement tray (301) has a number of clips (303) evenly distributed, and the clips (303) are symmetrically clipped onto the outside of the catalyst tube (302).

6. The multiplex primer kit for enriching gene target regions according to claim 5, characterized in that: The top of the placement tray (301) is provided with a placement groove (307), and the two ends of the inner side of the placement groove (307) are symmetrically fixed with limiting protrusions (312), and the limiting protrusions (312) are located at both ends of the microporous plate (308).

7. The multiplex primer kit for enriching gene target regions according to claim 1, characterized in that: The pressure plate mechanism (4) further includes a pressure plate (401), on which a plurality of first pressure blocks (402) and second pressure blocks (403) are evenly distributed, and a pressure groove (404) is provided on the pressure plate (401).

8. The multiplex primer kit for enriching gene target regions according to claim 1, characterized in that: A hinge (103) is movably mounted on the side of the lid (101), and the hinge (103) is connected to the side of the base (102).

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