PCR (Polymerase Chain Reaction) sealed detection box
By designing PCR sealing detection box, using embedded sealing grooves and guide holes to achieve automated sealing, the problems of cumbersome manual operation and difficulty in automatic sealing in the existing technology are solved, and the automation and multi-sample detection of PCR detection are realized, improving the Test efficiency and accuracy.
Patent Information
- Application Number
- CN202420974617.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-05-08
- Publication Date
- 2024-12-31
- Estimated Expiration
- 2034-05-08
AI Technical Summary
The existing PCR detection technology requires manual tedious sample processing and sealing operations, resulting in high requirements for the detection environment and technicians, and lack of automated capping methods, especially when the sample size is small, it is difficult to achieve automated inspection.
A PCR sealed detection box is designed, including the PCR microfluidic chip body and chip pallet. It can be automatically sealed through embedded sealant grooves and guide holes. It is equipped in an automated PCR detection equipment to realize sample sampling, nucleic acid extraction, and reagents. An automated process for formulation and real-time fluorescence PCR detection.
It realizes the automation and multi-sample detection of PCR detection, reduces the tedious work of professional technicians, improves detection efficiency and accuracy, and is suitable for integration into automated PCR detection equipment to ensure sealing effect and detection stability.
Smart Images
Figure CN222274571U_ABST
Abstract
Description
Technical Field
[0001] The utility model belongs to the technical field of biomedical detection, and in particular relates to a PCR sealed detection box. Background Art
[0002] Currently, when pathogen detection is carried out, samples are sent to the laboratory where professional technicians use large instruments to perform PCR (polymerase chain reaction) reactions, including independent extraction and purification and independent testing. Manual liquid preparation, sample addition, mixing, and machine loading are required. The work is cumbersome and has high requirements for the testing environment and technicians. In addition, sample contamination is prone to occur, which reduces the accuracy of the results.
[0003] In addition, commonly used PCR amplification carriers are generally 96-well plates, which can achieve automatic film coating and sealing. However, when the number of samples is relatively small, using a single PCR tube or eight-well tube requires manual cover and closure, and there is currently no good automated sealing method. When using microfluidic chips as reaction carriers and detection containers, there are currently the following sealing methods: (1) with a lid that requires manual sealing; (2) sticking items similar to tape to seal the sample inlet; (3) integrated card box with built-in sealing function. The first two methods are highly dependent on manual operation and cannot be used in automated products. The third method is mostly used for the detection of a single sample, with low detection throughput, and the number of targets detected at a single time is also limited.
[0004] Therefore, there is a need for a PCR detection box that can be integrated into an automated PCR detection device to achieve automatic sample loading, automatic sealing, and automatic detection. Utility Model Content
[0005] In order to solve at least one of the above technical problems, the utility model provides a PCR sealed detection box, which is installed in a PCR integrated automatic detection device. The user only needs to put the sample and consumables into the specified position of the detection device, and the detection device can automatically complete the whole process of sample sampling, nucleic acid extraction, reagent preparation, adding samples to the chip, sealing the chip, real-time fluorescence PCR detection, etc., and automatic detection of multiple samples can be achieved at a single time.
[0006] A PCR sealed detection box comprises a PCR microfluidic chip body and a chip support plate, wherein the PCR microfluidic chip body comprises a base and a cover sheet which are bonded to each other, wherein at least one reaction chamber is formed on the base, and injection holes are respectively formed on the cover sheet at the end position relative to each reaction chamber; a groove is provided on the second surface of the chip support plate as a chip reserved area, and the PCR microfluidic chip body is embedded in and fixed in the chip reserved area with a matching size.
[0007] Furthermore, at least one guide hole is formed on the first surface of the chip support plate, and the guide hole is aligned one by one with the injection holes of the PCR microfluidic chip body.
[0008] Furthermore, the chip support plate also includes at least one sealant groove, which is opened between the at least one guide hole and the corresponding injection hole, and is respectively connected to the corresponding guide hole and injection hole, for accommodating the sealant added from the guide hole to seal the injection hole.
[0009] Furthermore, the chip support plate further comprises an observation window, and the observation window is opened in a region of the first surface of the chip support plate corresponding to the reaction chamber of the PCR microfluidic chip body.
[0010] Furthermore, the chip tray also includes a tray mounting position identification pad, which is used to position the PCR sealed detection box with the PCR detection device when loading it into the PCR detection device.
[0011] Furthermore, the support plate mounting position identification pad is positioned by magnetic force.
[0012] Furthermore, the at least one sealant groove at one end of the at least one reaction chamber is a connected groove, rather than a separate sealant groove corresponding to each guide hole. This can increase the coverage area of the sealant, increase the contact area between the sealant and the surface of the PCR microfluidic chip body, make the seal of the sealant more secure, and avoid leakage.
[0013] Furthermore, a plurality of the grooves are arranged in an array on the chip support plate as the chip reserved area, and a PCR microfluidic chip body is embedded in each of the chip reserved areas.
[0014] Furthermore, the groove depth of the chip reserved area is smaller than the thickness of the PCR microfluidic chip body, so that after the PCR microfluidic chip body is loaded into the groove from the second surface of the chip support plate, the bottom of the PCR microfluidic chip body protrudes from the second surface of the chip support plate, and the bottom of the PCR sealed detection box is the bottom of the PCR microfluidic chip body. Maintaining a flat bottom structure is more conducive to cyclic heating during PCR detection, i.e., there is no need to additionally design a special heating plate in the PCR detection equipment to adapt to the uneven bottom.
[0015] Compared with the prior art, the beneficial effects of the utility model are:
[0016] (1) The PCR sealed detection box provided by the utility model can facilitate users to package and use the main body of the PCR microfluidic chip, and is suitable for manual sample addition and automated PCR detection microfluidic chips. The PCR microfluidic chip can be used alone or in combination with multiple chips. It can be placed on a matching PCR instrument for detection after manual sample addition by the user, or it can be used with a fully automatic detection device to solve the problems of automatic sample addition, automatic sealing, and automatic detection in POCT fully automatic instant detection. The user only needs to put the PCR microfluidic chip into the chip tray, and put the chip tray into the fixed position of the PCR detection device, and the detection device can automatically complete the entire process of sample sampling, nucleic acid extraction, reagent preparation, sample addition to the chip, sealing the chip, and real-time fluorescence PCR detection. It can realize the automatic detection of multiple samples at a time, reducing the tedious work of professional technicians such as sample addition and sealing. The PCR sealed detection box provided by the utility model can realize PCR fully automatic instant extraction and fluorescence detection;
[0017] (2) The PCR sealed detection box provided by the utility model simplifies and automates the sample loading of the PCR microfluidic chip, and is more suitable for integration into automated PCR detection equipment;
[0018] (3) The embodiment of the utility model provides a groove on the back side (second surface) of the chip support plate, so that the PCR microfluidic chip body is embedded and fixed in the groove by, for example, gluing in a size-adapted manner, so that the entire PCR sealed detection box becomes one, and the bottom of the PCR sealed detection box is the bottom of the PCR microfluidic chip body. Maintaining a flat bottom structure is more conducive to cyclic heating during the PCR detection process, that is, there is no need to additionally design a special heating plate in the PCR detection device to adapt to the uneven bottom;
[0019] (4) The embodiment of the utility model corrects the error of the pipette tip by setting a guide hole for the pipette tip of the automatic pipetting device on the chip support plate, thereby avoiding the situation where the pipette tip of the automatic pipetting device is not aligned with the injection port of the PCR microfluidic chip when adding samples to the PCR microfluidic chip, thereby ensuring the stability of the injection and the leakage-free addition of the sample;
[0020] (5) The embodiment of the utility model provides a hidden sealant groove in the chip support plate to store the sealant, thereby ensuring the stable sealing performance of the sealant after the PCR microfluidic chip is loaded with samples, thereby achieving a sealing effect;
[0021] (6) The embodiment of the utility model designs an expandable PCR microfluidic chip array on the chip support plate, so that multiple PCR microfluidic chips can be loaded at the same time, thereby realizing high-throughput detection of PCR microfluidic chips. BRIEF DESCRIPTION OF THE DRAWINGS
[0022] Figure 1 It is a schematic diagram of the top view of the PCR sealed detection box of Example 1 of the utility model;
[0023] Figure 2 It is a schematic cross-sectional view of the PCR sealed detection box of Example 1 of the utility model;
[0024] Figure 3 This is a schematic diagram of the structure explosion of the PCR sealed detection box of Example 1 of the utility model;
[0025] Figure 4 It is a schematic diagram of the back structure of the chip support plate of the PCR sealed detection box in Example 1 of the utility model;
[0026] Figure 5 It is a structural schematic diagram of a PCR sealed detection box with a microfluidic chip array according to Example 2 of the utility model.
[0027] In the figure:
[0028] 1-PCR microfluidic chip body; 2-chip support plate; 3-reaction chamber; 4-injection hole; 5-chip reserved area; 7-guide hole; 8-sealing glue groove; 9-observation window; 10-support plate installation position identification pad. DETAILED DESCRIPTION
[0029] The utility model is further described below in conjunction with the accompanying drawings.
[0030] Example 1
[0031] The utility model provides a PCR sealed detection box. Figure 1-2 They are respectively a top view structural schematic diagram and a cross-sectional structural schematic diagram of the PCR sealed detection box of Example 1 of the utility model. Figure 3 Schematic diagram of the structure explosion of the PCR sealed detection box of Example 1 of the utility model. Figure 1-3 As shown, the PCR sealed detection box includes a PCR microfluidic chip body 1 and a chip support plate 2. The PCR microfluidic chip body 1 is formed by laminating (for example, laminating by bonding) a substrate (a silicon substrate in this embodiment) and a cover sheet (a glass slide in this embodiment) and then cutting. A plurality of grooves are formed on the silicon substrate by etching as reaction chambers 3, and injection holes 4 are formed on the glass slide at both ends relative to each reaction chamber 3 by, for example, laser drilling. In this embodiment, the chip support plate 2 can be a PC (polycarbonate) or PE (polyethylene) polymer material. Figure 4 Schematic diagram of the back structure of the chip support plate of the PCR sealed detection box of the utility model embodiment 1. Figure 4As shown, the second surface of the chip support plate 2 is provided with a groove as a chip reserved area 5, and the PCR microfluidic chip body 1 is embedded and fixed in the chip reserved area 5 with a size adaptation. In the present embodiment, the frame size of the chip reserved area 5 is adapted to the size of the PCR microfluidic chip body 1, so that the PCR microfluidic chip body 1 can be just embedded in the groove of the chip reserved area 5, and the microfluidic chip body 1 is fixed in the groove by dispensing glue at the four corners of the groove. Preferably, the four corners of the groove of the chip reserved area 5 are designed to be rounded, which is more conducive to dispensing glue. By providing a groove on the back side (second surface) of the chip support plate 2 to embed the PCR microfluidic chip body 1, the entire PCR sealed detection box is integrated, and the bottom of the PCR sealed detection box is the bottom of the PCR microfluidic chip body 1, and the structure of maintaining a flat bottom is more conducive to cyclic heating during the PCR detection process, that is, there is no need to additionally design a special heating plate that adapts to the uneven bottom in the PCR detection device.
[0032] like Figure 1 , Figure 2 , Figure 4 As shown, the front side (first surface) of the chip support plate 2 is provided with a plurality of guide holes 7, and the guide holes 7 are aligned one by one with the injection holes 4 of the PCR microfluidic chip body 1 in the direction perpendicular to the first and second surfaces of the chip support plate 2, that is, the number of the guide holes 7 corresponds to the number of the injection holes 4. Below the guide holes 7, that is, between the guide holes 7 and the injection holes 4, a sealant groove 8 is also provided, and the sealant groove 8 is respectively connected with the guide holes 7 and the injection holes 4 in the direction perpendicular to the first and second surfaces of the chip support plate 2, and is used to accommodate the sealant added from the guide holes 7 by the pipette tip of the pipetting device to seal the injection holes 4. Generally speaking, the diameter of the guide hole 7 is slightly larger than the size of the pipette tip of the pipetting device, which is convenient for fixing the pipette tip to ensure that the front end of the pipette tip can be aligned with the injection hole 4, and can be vertically pressed above the injection hole 4 to ensure that the sample is added without leakage. By setting a guide hole 7 for the pipette tip of the automatic pipetting device on the chip support plate 2, the error of the pipette tip is corrected, thereby avoiding the situation where the pipette tip of the automatic pipetting device is not aligned with the PCR microfluidic chip injection port 4 when adding samples to the PCR microfluidic chip body 1, thereby ensuring the stability of injection.
[0033] Preferably, Figure 4 As shown, the sealant groove 8 is designed as a connected groove at one end of the multiple parallel reaction chambers 3, rather than a separate sealant groove corresponding to each guide hole 7. This can increase the coverage area of the sealant, increase the contact area between the sealant and the surface of the PCR microfluidic chip body 1, make the seal of the sealant more secure, and avoid leakage. The embodiment of the utility model provides a hidden sealant groove 8 in the chip support plate 2 to store the sealant, ensure that the sealing performance of the sealant after the PCR microfluidic chip body 1 is added, and achieve a sealing effect.
[0034] Further, such as Figure 2 As shown, an observation window 9 is also provided on the chip support plate 2, and the observation window 9 is provided on the front side of the chip support plate 2 corresponding to the area of the reaction chamber 3 of the PCR microfluidic chip body 1. That is to say, in this embodiment, the area on the back side of the chip support plate 2 where the PCR microfluidic chip body 1 is embedded is hollowed out, and when the PCR microfluidic chip body 1 is embedded into the chip support plate 2 and fixed by glue, the injection holes 4 on the PCR microfluidic chip body 1 are respectively aligned one by one with the guide holes 7 of the chip support plate 2, and the reaction chamber 3 area is just located in the hollowed-out area (observation window 9) and exposed, so that it is convenient to obtain fluorescence detection data from the observation window 9.
[0035] Further, such as Figure 1 As shown, the chip tray 2 also includes a tray installation position identification pad 10, which is used to position the PCR sealed detection box with the PCR detection device when it is loaded into the PCR detection device. For example, in this embodiment, the tray installation position identification pads 10 are respectively set at the four corners of the chip tray 2. Specifically, the tray installation position identification pads 10 can be positioned by magnetic force. The tray installation position identification pads 10 are made of metal iron, for example, and are automatically positioned by magnetic attraction with a magnetic sheet set at a specific position on the PCR detection device.
[0036] The working process of the PCR sealed detection box provided by the embodiment of the utility model is as follows: first, the PCR sealed detection box (the observation window 9 on the chip support plate 2 faces upward) is loaded onto the adapted PCR detection device; the PCR microfluidic chip body is automatically fixed under the adsorption force of the PCR detection device, and the PCR microfluidic chip body 1 is completely fitted with the heating position of the PCR detection device, which facilitates the conduction of heat. When using magnetic positioning, magnets at corresponding positions are reserved on the PCR detection device, and the support plate mounting position identification pad 10 can be adsorbed and aligned on the POCT integrated machine at the same time through the magnetic sheet 11. According to the program flow, the pipetting robot on the PCR detection device grabs the adapted gun tip, absorbs the prepared PCR detection reagent, and the gun tip is aligned with the injection hole 4 of the PCR microfluidic chip body 1 through the gun tip guide hole 7. Because some gun tips are bent and not completely vertical, there will be a position error between them and the sampling hole 4. The error can be corrected through the guide hole 7, and pressure is applied through the pipette to make the front end of the gun tip fit tightly with the sampling hole 4 of the PCR microfluidic chip body 1. The internal air pressure of the pipette pushes the liquid in the gun tip to flow into the PCR microfluidic chip body 1 to complete the sample addition; after the sample addition, the gun tip is replaced again, and after absorbing the sealant, the gun tip dispenses the sealant through the guide hole 7 to the sealant groove 8 below to seal the sampling hole 4 of the PCR microfluidic chip body 1, and then the sealant is cured by ultraviolet light to make the PCR microfluidic chip body 1 completely sealed; after sealing, start the PCR amplification test, obtain the fluorescence detection data through the observation window 9 on the chip support plate 2, complete the data analysis, and get the test results.
[0037] Example 2
[0038] Figure 5 Schematic diagram of the structure of the PCR sealed detection box with microfluidic chip array according to Example 2 of the utility model. In this embodiment, a plurality of PCR microfluidic chip bodies 1 are arranged in an array on the chip support plate 2. For example, Figure 5 As shown in the quadruple (2*2 array) and octacouple (4*2 array), in the octacouple arrangement, a total of 8 3-channel PCR microfluidic chip bodies 1 can realize PCR detection of 24 samples.
[0039] Finally, it should be noted that the above description is only a preferred embodiment of the utility model and is not intended to limit the utility model. Although the utility model is described in detail with reference to the above embodiments, those skilled in the art can still modify the technical solutions recorded in the above embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the utility model shall be included in the protection scope of the utility model.
Claims
1. A PCR sealed detection kit, characterized in that: The invention comprises a PCR microfluidic chip body (1) and a chip support plate (2), wherein the PCR microfluidic chip body (1) comprises a base and a cover plate which are bonded to each other, wherein at least one reaction chamber (3) is formed on the base, and injection holes (4) are respectively formed on the cover plate at the end position relative to each reaction chamber (3); a groove is provided on the second surface of the chip support plate (2) as a chip reserved area (5), and the PCR microfluidic chip body (1) is embedded in and fixed in the chip reserved area (5) with a size that matches the size.
2. The PCR sealed detection kit according to claim 1, characterized in that: The first surface of the chip support plate (2) is provided with at least one guide hole (7), and the guide hole (7) is aligned one by one with the injection holes (4) of the PCR microfluidic chip body (1).
3. The PCR sealed detection kit according to claim 2, characterized in that: The chip support plate (2) further comprises at least one sealing glue groove (8), wherein the at least one sealing glue groove (8) is arranged between the at least one guide hole (7) and the corresponding injection hole (4), and is respectively connected to the corresponding guide hole (7) and the injection hole (4), and is used for accommodating the sealing glue added from the guide hole (7) to seal the injection hole (4).
4. The PCR sealed detection kit according to claim 2, characterized in that: The chip support plate (2) further comprises an observation window (9), wherein the observation window (9) is provided on the first surface of the chip support plate (2) in an area corresponding to the reaction chamber (3) of the PCR microfluidic chip body (1).
5. The PCR sealed detection kit according to claim 1, characterized in that: The chip support plate (2) further comprises a support plate installation position identification pad (10) for positioning the PCR sealed detection box with the PCR detection device when the PCR sealed detection box is loaded into the PCR detection device.
6. The PCR sealed detection kit according to claim 5, characterized in that: The support plate installation position identification pad (10) is positioned by magnetic force.
7. The PCR sealed detection kit according to claim 3, characterized in that: The at least one sealant groove (8) is a connected groove at one end of the at least one reaction chamber (3).
8. The PCR sealed detection kit according to claim 1, characterized in that: A plurality of the grooves are arranged in an array on the chip support plate (2) as the chip reserved area (5), and a PCR microfluidic chip body (1) is embedded in each of the chip reserved areas (5).
9. The PCR sealed detection kit according to claim 1, characterized in that: The groove depth of the chip reserved area (5) is less than the thickness of the PCR microfluidic chip body (1).