PCR test assembly and system
By designing a PCR test component containing magnetic parts, the problem of residual magnetic beads interfering with fluorescence detection in the PCR consumable board was solved, and the normality of the amplification curve and the accuracy of the test were achieved.
Patent Information
- Application Number
- CN202421244825.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-06-03
- Publication Date
- 2025-05-02
- Estimated Expiration
- 2034-06-03
AI Technical Summary
After the magnetic bead extraction of existing PCR consumable plates, some magnetic beads are often left in the consumable plate due to sufficient extraction. In subsequent fluorescence detection, the deposition of magnetic beads blocks the LED excitation light, resulting in abnormal amplification curve.
A PCR test assembly was designed, including a housing plate, a sample plate, a PCR tube and a magnetic component. After extracting the target substance in the PCR tube through magnetic beads, a small amount of residual magnetic beads are adsorbed to the side wall of the PCR tube using magnetic parts, thereby avoiding the magnetic beads from blocking the fluorescent signal.
It effectively avoids interference from magnetic beads in fluorescence detection, ensures the normality of the amplification curve, and improves the accuracy of PCR tests.
Smart Images
Figure CN222821566U_ABST
Abstract
Description
Technical Field
[0001] The present application relates to the field of in vitro detection technology, and in particular to PCR test components and systems. Background Art
[0002] PCR (Polymerase Chain Reaction) is a molecular biology technique used to amplify specific DNA fragments. It can be seen as a special DNA replication outside the body. The biggest feature of PCR is that it can greatly increase trace amounts of DNA. Therefore, whether it is ancient organisms in fossils, remains of historical figures, or hair, skin or blood left by murderers in cases decades ago, as long as a tiny bit of DNA can be separated, it can be amplified by PCR and compared. This is also the power of "trace evidence".
[0003] In vitro diagnosis is an important part of clinical medicine, and molecular diagnosis covered by in vitro diagnosis is an indispensable part of clinical medicine. Molecular diagnosis refers to the technology of using molecular biology methods to detect changes in the structure or expression level of genetic material in the patient's body to make a diagnosis. It is the main method of predictive diagnosis. It can be used for diagnosis of individual genetic diseases and prenatal diagnosis. The first step of molecular diagnosis is to extract nucleic acids, which is usually achieved through PCR technology. The magnetic bead method is often used in the extraction process.
[0004] However, after the magnetic bead extraction step is completed, some magnetic beads often remain in the existing PCR consumables due to insufficient extraction. In the subsequent fluorescence detection process, the magnetic beads are easily deposited at the bottom of the sample in the consumables, causing the LED excitation light that hits the sample from the bottom to be blocked by the residual magnetic beads, thereby causing interference in the fluorescence detection and causing abnormal amplification curves. Utility Model Content
[0005] Based on this, it is necessary to provide a PCR test component and system in view of the fact that after the existing PCR consumables plate completes the magnetic bead extraction step, some magnetic beads often remain in the consumables plate due to insufficient extraction, and in the subsequent fluorescence detection process, the magnetic beads are easily deposited at the bottom of the sample in the consumables plate, causing the LED excitation light that hits the sample from the bottom to be blocked by the residual magnetic beads, thereby causing interference in the fluorescence detection and leading to abnormal amplification curves.
[0006] A PCR test kit, comprising:
[0007] A receiving plate, wherein the receiving plate is provided with a plurality of through holes and a plurality of receiving grooves along the thickness direction thereof, the plurality of receiving grooves are located on the same side of the receiving plate, and the through holes are gradually reduced in a direction close to the receiving grooves along the thickness direction of the receiving plate;
[0008] A sample plate, wherein the sample plate is provided with a plurality of sample holes, the plurality of sample holes correspond to the plurality of through holes one by one, and the sample plate covers the containing plate;
[0009] A plurality of PCR tubes, each having an open end and a closed end at both ends, the PCR tubes gradually shrinking from the open end to the closed end along the axial direction thereof, the plurality of PCR tubes being inserted into the sample wells one by one, and the positions of the open end and the sample wells being relatively fixed, the PCR tubes being adapted to the through holes;
[0010] A plurality of magnetic members are located in the plurality of receiving grooves in a one-to-one correspondence, and the positions of the magnetic members and the receiving plate are relatively fixed.
[0011] During the specific use of the above-mentioned PCR test assembly, after the target substance in the PCR tube is extracted by magnetic beads, the sample plate is covered on one side of the through hole of the receiving plate, and the PCR tubes are inserted into the corresponding through holes one by one. Due to the presence of the magnetic component, a small amount of magnetic beads remaining in the sample in the PCR tube are adsorbed from the bottom of the PCR tube to the side wall of the PCR tube, so that a light guide can be used to emit a fluorescent signal to the PCR tube through the opening on the side of the through hole away from the sample plate. The fluorescent signal can fully pass through the sample in the PCR tube and generate a fluorescent signal reflected back to the light guide, thereby avoiding interference in fluorescence detection and ensuring a normal amplification curve.
[0012] In one embodiment, the PCR test assembly further includes a constant temperature module, and the constant temperature module is disposed on the containing plate to ensure that the temperature of the containing plate is constant.
[0013] In one embodiment, the PCR test assembly further includes a plurality of fasteners, and the plurality of fasteners are connected to the plurality of receiving slots in a one-to-one correspondence, so that the magnetic member receiving slot is located inside the receiving slot and is fixed relative to the position of the receiving slot.
[0014] In one embodiment, the accommodating groove includes a first groove and a second groove connected to each other, the inner diameter of the first groove is smaller than the inner diameter of the second groove, the first groove is located on a side of the second groove away from the opening of the accommodating groove, and the magnetic member is relatively fixed in position to the first groove.
[0015] In one embodiment, the fastener includes a connecting portion and a connecting head;
[0016] The connecting portion is threadedly connected to the inner wall of the first groove, the magnetic member is located between the connecting portion and the bottom wall of the first groove, and the connecting head is located in the second groove; or
[0017] The magnetic member is annular, the outer wall of the magnetic member is threadedly connected to the inner wall of the first groove, the connecting portion is threadedly connected to the inner wall of the magnetic member, and the connecting head is located in the second groove.
[0018] In one embodiment, the PCR test assembly further includes a cover tube, the sample plate covers an opening at one end of the cover tube, the outer periphery of the sample plate is connected to the inner periphery of the cover tube, the cover tube is covered on the outer periphery of the containing plate, and the cover tube is adapted to the containing plate.
[0019] In one embodiment, the housing cylinder and the sample plate form a 96-well full skirt plate.
[0020] In one embodiment, the fastener is a bolt or a screw.
[0021] An embodiment of the present application further provides a PCR test system, the PCR test system comprising: a temperature control platform and the PCR test component;
[0022] A side of the receiving plate facing away from the sample plate is in contact with the temperature control platform.
[0023] In one embodiment, the PCR test system further includes a plurality of light guides, which are disposed on the surface of the temperature control platform. The plurality of light guides correspond one-to-one to the plurality of through holes and are used to guide optical signals into or out of the through holes.
[0024] During the specific use of the above-mentioned PCR test system, after the target substance in the PCR tube is extracted by magnetic beads, the sample plate is covered on one side of the through hole of the receiving plate, and the PCR tubes are inserted into the corresponding through holes one by one. Due to the presence of the magnetic component, a small amount of magnetic beads remaining in the sample in the PCR tube are adsorbed from the bottom of the PCR tube to the side wall of the PCR tube, so that the light guide can be used to emit a fluorescent signal to the PCR tube through the opening on the side of the through hole away from the sample plate. The fluorescent signal can fully pass through the sample in the PCR tube and generate a fluorescent signal reflected back to the light guide, thereby avoiding interference in the fluorescence detection and ensuring that the amplification curve is normal. At the same time, the temperature control platform can control the temperature according to different test samples and experimental processes to ensure that the test can proceed smoothly. BRIEF DESCRIPTION OF THE DRAWINGS
[0025] Figure 1 A schematic diagram of a receiving plate in a PCR test assembly according to an embodiment.
[0026] Figure 2 FIG. 1 is a schematic diagram of a sample plate and PCR tubes in a PCR test assembly according to an embodiment.
[0027] Figure 3 for Figure 2 Enlarged view of point A in the middle.
[0028] Figure 4 FIG. 4 is a partial schematic diagram of a PCR test system according to an embodiment of the present invention.
[0029] Description of Figure Numbers:
[0030] 100-PCR test kit;
[0031] 110 - receiving plate; 111 - through hole; 112 - receiving groove; 113 - first groove; 114 - second groove;
[0032] 120-96-well full skirt plate; 121-sample plate; 122-PCR tube; 123-sample well; 124-open end; 125-closed end; 126-shield;
[0033] 130-magnetic part; 131-constant temperature module; 132-fastener; 133-connecting part; 134-connecting head;
[0034] 200-PCR test system;
[0035] 210-temperature control platform; 211-light guide;
[0036] 220-Magnetic beads. DETAILED DESCRIPTION
[0037] In order to make the above-mentioned purposes, features and advantages of the present application more obvious and easy to understand, the specific implementation methods of the present application are described in detail below in conjunction with the accompanying drawings. In the following description, many specific details are set forth to facilitate a full understanding of the present application. However, the present application can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present application, so the present application is not limited by the specific embodiments disclosed below.
[0038] In the description of the present application, it should be understood that if the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise", "axial", "radial", "circumferential", etc. appear, the orientation or position relationship indicated by these terms is based on the orientation or position relationship shown in the accompanying drawings, which is only for the convenience of describing the present application and simplifying the description, and does not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present application.
[0039] In addition, if the terms "first" or "second" appear, these terms are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include at least one of the features. In the description of this application, if the term "plurality" appears, the meaning of "plurality" is at least two, such as two, three, etc., unless otherwise clearly and specifically defined.
[0040] In this application, unless otherwise clearly specified and limited, if the terms "installed", "connected", "connected", "fixed" and the like appear, these terms should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integrated connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements, unless otherwise clearly defined. For ordinary technicians in this field, the specific meanings of the above terms in this application can be understood according to the specific circumstances.
[0041] In the present application, unless otherwise clearly specified and limited, if there is a description that a first feature is "above" or "below" a second feature, etc., or similar descriptions appear, it may mean that the first and second features are in direct contact, or that the first and second features are in indirect contact through an intermediate medium. Moreover, the first feature being "above", "above" and "above" the second feature may mean that the first feature is directly above or obliquely above the second feature, or simply means that the first feature is higher in level than the second feature. The first feature being "below", "below" and "below" the second feature may mean that the first feature is directly below or obliquely below the second feature, or simply means that the first feature is lower in level than the second feature.
[0042] It should be noted that if an element is referred to as being "fixed to" or "disposed on" another element, it may be directly on the other element or there may be a central element. If an element is considered to be "connected to" another element, it may be directly connected to the other element or there may be a central element at the same time. If any, the terms "vertical", "horizontal", "upper", "lower", "left", "right" and similar expressions used in this application are for illustrative purposes only and do not represent the only implementation method.
[0043] See also Figure 1 and Figure 2 , Figure 1 FIG. 1 is a schematic diagram showing a receiving plate 110 in a PCR test assembly 100 in an embodiment of the present application. Figure 2A schematic diagram of a sample plate 121 in a PCR test assembly 100 in an embodiment of the present application is shown. The PCR test assembly 100 provided in an embodiment of the present application includes: a receiving plate 110, a sample plate 121, a plurality of PCR tubes 122 and a plurality of magnetic members 130.
[0044] In the above-mentioned PCR test assembly 100, the containing plate 110 is provided with a plurality of through holes 111 and a plurality of containing grooves 112 along its thickness direction, and the through holes 111 are gradually reduced in the direction close to the containing grooves 112 along the thickness direction of the containing plate 110; the sample plate 121 is provided with a plurality of sample holes 123, and the plurality of sample holes 123 correspond to the plurality of through holes 111 one by one, and the sample plate 121 covers the containing plate 110; the two ends of the PCR tube 122 are respectively an open end 124 and a closed end 125, and the PCR tube 122 is gradually reduced from the open end 124 to the closed end 125 along its axial direction, and the plurality of PCR tubes 122 are inserted into the sample holes 123 one by one, and the positions of the open ends 124 and the sample holes 123 are relatively fixed, and the PCR tubes 122 are adapted to the through holes 111; the plurality of magnetic members 130 are located in the plurality of containing grooves 112 one by one, and the positions of the magnetic members 130 and the containing plate 110 are relatively fixed.
[0045] See also Figure 1 and Figure 2 and Figure 4 In the specific use of the PCR test assembly 100, after the target substance in the PCR tube 122 is extracted by the magnetic beads 220, the sample plate 121 covers the side of the receiving plate 110 where the through holes 111 are opened, and the PCR tubes 122 are inserted into the corresponding through holes 111 one by one. Due to the presence of the magnetic member 130, a small amount of magnetic beads 220 remaining in the sample in the PCR tube 122 are removed from the bottom of the PCR tube 122 (such as Figure 3 ) is adsorbed to the side wall of the PCR tube 122 (e.g. Figure 4 ), so that the light guide 211 can be used to emit a fluorescent signal to the PCR tube 122 through the opening on the side of the through hole 111 away from the sample plate 121. The fluorescent signal can fully pass through the sample in the PCR tube 122 and generate a fluorescent signal reflected back to the light guide 211, thereby avoiding interference in the fluorescence detection and ensuring that the amplification curve is normal.
[0046] Specifically, the sample plate covers the side of the containing plate where no containing groove is provided, that is, the side where the inner diameter of the through hole is larger.
[0047] Specifically, each receiving groove 112 is adjacent to the corresponding through hole 111, and each receiving groove 112 is located in the same direction of the corresponding receiving hole, so that the magnetic beads 220 are adsorbed in the same direction of the PCR tube 122, making the adsorption of the magnetic beads 220 more uniform.
[0048] In one embodiment, the PCR test component 100 further includes a constant temperature module 131, which is disposed on the containing plate 110 to control the temperature of the containing plate 110, so that the containing plate 110 can be temperature-controlled by the constant temperature module 131 according to different test conditions. Since the through hole 111 is adapted to the PCR tube 122, the sample in the PCR tube 122 can be temperature-controlled.
[0049] In one embodiment, the PCR test assembly 100 further includes a plurality of fasteners 132 , which are connected one-to-one with the plurality of receiving slots 112 , so that the magnetic member 130 receiving slot 112 is located in the receiving slot 112 and is relatively fixed with respect to the position of the receiving slot 112 .
[0050] In one embodiment, the receiving groove 112 includes a first groove 113 and a second groove 114 connected to each other, the inner diameter of the first groove 113 is smaller than the inner diameter of the second groove 114, the first groove 113 is located on the side of the second groove 114 away from the opening of the receiving groove 112, and the position of the magnetic member 130 and the first groove 113 are relatively fixed, so that the fastener 132 can be fastened from the second groove 114 with a larger inner diameter, thereby facilitating the fastening process.
[0051] In one embodiment, the fastener 132 includes a connecting portion 133 and a connecting head 134; the connecting portion 133 is threadedly connected to the inner wall of the first groove 113, the magnetic member 130 is located between the connecting portion 133 and the bottom wall of the first groove 113, and the connecting head 134 is located in the second groove 114, so that the magnetic member 130 is fixed in the first groove 113 through the connecting portion 133 of the fastener 132.
[0052] In another embodiment, the connecting portion 133 is located in the first groove 113 , the magnetic member 130 is located between the connecting portion 133 and the bottom wall of the first groove 113 , and the connecting head 134 is threadedly connected to the inner wall of the second groove 114 .
[0053] In another embodiment, the magnetic member 130 is annular, the outer wall of the magnetic member 130 is threadedly connected to the inner wall of the first groove 113, the connecting portion 133 is threadedly connected to the inner wall of the magnetic member 130, and the connecting head 134 is located in the second groove 114, so that the magnetic member 130 can generate magnetic attraction to the magnetic beads 220 along the axial direction of the first groove 113, so that the magnetic beads 220 can be attached to the side wall of the PCR tube 122 in a larger area, ensuring that the magnetic beads 220 will not bulge due to aggregation and affect the fluorescence detection.
[0054] In one embodiment, the PCR test assembly 100 also includes a cover tube 126, the sample plate 121 covers an opening at one end of the cover tube 126, the outer periphery of the sample plate 121 is connected to the inner periphery of the cover tube 126, the cover tube 126 is covered on the outer periphery of the containing plate 110, and the cover tube 126 is adapted to the containing plate 110, thereby facilitating the relative fixation of the sample plate 121 and the containing plate 110.
[0055] In one embodiment, the PCR test assembly 100 , the housing cylinder 126 and the sample plate 121 form a 96-well full skirt plate 120 .
[0056] Specifically, the 96-well full skirt plate 120 and the PCR tube 122 are referred to in the prior art and will not be described in detail.
[0057] In one embodiment, in the PCR test assembly 100, the fastener 132 is a bolt or a screw, and may also be other components, which will not be described in detail.
[0058] An embodiment of the present application further provides a PCR test system 200 , which includes: a temperature control platform 210 and a PCR test component 100 ;
[0059] The side of the receiving plate 110 facing away from the sample plate 121 is in contact with the temperature-controlled platform 210 .
[0060] During the specific use of the above-mentioned PCR test system 200, after the target substance in the PCR tube 122 is extracted by the magnetic beads 220, the sample plate 121 covers the side of the through hole 111 of the receiving plate 110, and the PCR tubes 122 are inserted into the corresponding through holes 111 one by one. Due to the presence of the magnetic member 130, a small amount of magnetic beads 220 remaining in the sample in the PCR tube 122 is adsorbed from the bottom of the PCR tube 122 to the side wall of the PCR tube 122, so that the light guide 211 can be used to emit a fluorescent signal to the PCR tube 122 through the opening on the side of the through hole 111 away from the sample plate 121. The fluorescent signal can fully pass through the sample in the PCR tube 122 and generate a fluorescent signal reflected back to the light guide 211, thereby avoiding interference in the fluorescence detection and ensuring that the amplification curve is normal. At the same time, the temperature control platform 210 can perform temperature control according to different test samples and experimental processes to ensure that the test can proceed smoothly.
[0061] In one embodiment, the PCR test system 200 also includes a plurality of light guides 211, which are disposed on the surface of the temperature control platform 210. The plurality of light guides 211 correspond one-to-one to the plurality of through holes 111, and are used to import or export optical signals into or out of the through holes 111, so that the light guides 211 can stably correspond one-to-one to the through holes 111, thereby facilitating the alignment process and the fluorescence signal collection process during the fluorescence detection process.
[0062] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.
[0063] The above-described embodiments only express several implementation methods of the present application, and the descriptions thereof are relatively specific and detailed, but they cannot be construed as limiting the scope of the patent application. It should be pointed out that, for a person of ordinary skill in the art, several variations and improvements can be made without departing from the concept of the present application, and these all belong to the protection scope of the present application. Therefore, the protection scope of the patent application shall be subject to the attached claims.
Claims
1. A PCR test kit, characterized in that: The PCR test kit includes: A receiving plate, wherein the receiving plate is provided with a plurality of through holes and a plurality of receiving grooves along the thickness direction thereof, the plurality of receiving grooves are located on the same side of the receiving plate, and the through holes are gradually reduced in a direction close to the receiving grooves along the thickness direction of the receiving plate; A sample plate, wherein the sample plate is provided with a plurality of sample holes, the plurality of sample holes correspond to the plurality of through holes one by one, and the sample plate covers the containing plate; A plurality of PCR tubes, each having an open end and a closed end at both ends, the PCR tubes gradually shrinking from the open end to the closed end along the axial direction thereof, the plurality of PCR tubes being inserted into the sample wells one by one, and the positions of the open end and the sample wells being relatively fixed, the PCR tubes being adapted to the through holes; A plurality of magnetic members are located in the plurality of receiving grooves in a one-to-one correspondence, and the positions of the magnetic members and the receiving plate are relatively fixed.
2. The PCR test kit according to claim 1, characterized in that: The PCR test assembly also includes a constant temperature module, which is arranged on the containing plate to ensure that the temperature of the containing plate is constant.
3. The PCR test kit according to claim 1, characterized in that: The PCR test assembly also includes a plurality of fasteners, and the plurality of fasteners are connected to the plurality of receiving slots in a one-to-one correspondence, so that the magnetic component receiving slot is located in the receiving slot and is fixed relative to the position of the receiving slot.
4. The PCR test kit according to claim 3, characterized in that: The receiving groove includes a first groove and a second groove connected to each other, the inner diameter of the first groove is smaller than the inner diameter of the second groove, the first groove is located on a side of the second groove away from the opening of the receiving groove, and the magnetic member is relatively fixed to the first groove.
5. The PCR test kit according to claim 4, characterized in that: The fastener comprises a connecting portion and a connecting head; The connecting portion is threadedly connected to the inner wall of the first groove, the magnetic member is located between the connecting portion and the bottom wall of the first groove, and the connecting head is located in the second groove; or The magnetic member is annular, the outer wall of the magnetic member is threadedly connected to the inner wall of the first groove, the connecting portion is threadedly connected to the inner wall of the magnetic member, and the connecting head is located in the second groove.
6. The PCR test kit according to claim 1, characterized in that: The PCR test assembly also includes a cover tube, the sample plate covers an opening at one end of the cover tube, the outer periphery of the sample plate is connected to the inner periphery of the cover tube, the cover tube is covered on the outer periphery of the receiving plate, and the cover tube is adapted to the receiving plate.
7. The PCR test kit according to claim 6, characterized in that: The cover cylinder and the sample plate form a 96-well full skirt plate.
8. The PCR test kit according to claim 3, characterized in that: The fastener is a bolt or a screw.
9. A PCR test system, characterized in that: The PCR test system comprises: a temperature control platform and a PCR test component according to any one of claims 1 to 8; A side of the receiving plate facing away from the sample plate is in contact with the temperature control platform.
10. The PCR test system according to claim 9, characterized in that: The PCR test system further comprises a plurality of light guides, which are arranged on the surface of the temperature control platform. The plurality of light guides correspond to the plurality of through holes one by one and are used to guide the optical signals into or out of the through holes.