Closed type sample treatment and detection card box device with pipetting function

Abstract

The invention discloses a closed sample treatment and detection card box device with a pipetting function, and relates to the technical field of closed sample treatment and detection.The closed sample treatment and detection card box device comprises a base and an upper cover, the upper cover is rotatably installed on the base, a cavity and a reaction detection pipe are distributed in the base around the center, and the reaction detection pipe is connected with the cavity. A reaction detection tube is arranged in the upper cover, a sleeve structure is arranged at the position, deviating from the center, of the upper cover, a pipette is installed in the sleeve structure in an up-down sliding mode, and a sealing structure used for sealing the reaction detection tube is arranged in the upper cover. Therefore, the environmental condition of the reaction detection area cannot be polluted by external air in the whole use process of the card box, the influence on the detection precision is avoided, and the temperature difference between the reaction hole pipe part and the top cover can be effectively avoided in the heating process, so that the environmental condition of reaction detection is optimized, and the detection efficiency is improved. And meanwhile, the outer diameter of the card box is reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of closed sample processing and detection, and particularly to a cartridge device for closed sample processing and detection with a pipetting function. Background Art

[0002] Sample analysis in the fields of biology, chemistry, and medicine generally includes steps such as sample preparation, reaction, separation, and detection. Conventional experimental operations are mostly carried out in an open experimental environment, where the sample can be in direct contact with the outside through the air. However, there are often some samples that are highly harmful to operators, easily interfered by the outside, or cause interference to the outside, such as biological samples containing pathogenic pathogens. In order to reduce biological hazards, there is an urgent need to invent a device with functions such as closed sample processing, reaction, and product detection;

[0003] Currently, cartridges for sample analysis usually use micro-pipes and micro-valves to achieve precise control of reagents or samples. However, the fluid control accuracy is usually affected by the characteristics of the sample or reagent, such as viscosity, surface tension, etc. Therefore, such cartridges are usually only applicable to a certain type or several types of reagents and do not have universal applicability;

[0004] For the amplification detection of nucleic acids, existing cartridge devices usually use a micro-valve method to achieve sealing, or use liquids such as silicone oil or paraffin for sealing to avoid sample evaporation due to high temperature. Both methods have differences in the sample reaction temperature from traditional PCR tubes and thermal lids. Therefore, it is necessary to optimize and improve the reagent system or reaction conditions according to the actual situation;

[0005] At the same time, the cartridge requires a relatively complex structure to achieve the horizontal and vertical movement of the pipette inside the closed box; the cartridge needs to have a sample addition port and a lid for sealing the sample addition port, etc., resulting in a complex structure or a large volume of the existing cartridge. Summary of the Invention

[0006] The purpose of the present invention is to provide a cartridge device for closed sample processing and detection with a pipetting function, which can be applied to various detection technologies including biochemical analysis, immunoassay, nucleic acid detection, etc., to solve the problems raised in the above background art.

[0007] To achieve the above purpose, the present invention provides the following technical solution: A cartridge device for closed sample processing and detection with a pipetting function, comprising a base and an upper cover. The upper cover is rotatably installed on the base. At least one chamber is distributed around the center in the base. Reaction and detection tubes are detachably arranged around the center at the bottom of the base. The upper cover is provided with a sleeve structure at a position deviating from the center. A pipette is slidably installed up and down in the sleeve structure. The upper cover is provided with a sealing structure for sealing the reaction and detection tubes.

[0008] Preferably, the sealing structure includes a guiding hole and a sealing cover arranged in the upper cover, and the guiding hole corresponds to the reaction detection tube.

[0009] Preferably, the sealing cover is a sealing film with elastic characteristics, and is fixedly connected to the bottom of the upper cover at the position area of the guiding hole.

[0010] Preferably, the sealing cover is a tube cover with a cylindrical structure, and the tube cover can slide up and down in the guiding hole.

[0011] Preferably, the tip at the bottom of the pipette is not on the same straight line as the central axis of the inner cavity on the upper side, and the eccentric distance is 3 mm.

[0012] Preferably, the bottom of the tip of the pipette has an anti-blocking structure, and the anti-blocking structure is a wedge-shaped structure.

[0013] Preferably, a positioning hole is arranged at the upper end of the pipette, and a positioning pin is arranged on the upper cover at the corresponding position, so as to achieve precise circumferential positioning of the pipette.

[0014] Preferably, a piston is slidably installed in the inner cavity of the pipette. A connecting groove is arranged in the middle of the upper end surface of the piston for connection and installation with an external connecting mechanism. The piston is in sealed contact with the side wall of the inner cavity and can move along the axis direction of the pipette to achieve liquid suction or liquid discharge.

[0015] Preferably, a first sealing ring is installed between the pipette and the side wall of the sleeve structure, and a second sealing ring is installed between the outer end surface of the base and the bottom cavity wall of the upper cover, and both are "O"-shaped sealing rings.

[0016] Preferably, both the first sealing ring and the second sealing ring are "O"-shaped sealing rings.

[0017] Preferably, the whole sealing cover is made of a transparent material, so as to detect the fluorescence signal of the reaction hole from above.

[0018] Preferably, there are elastic bayonets on the upper cover and the base. When the two are aligned, there is an obvious tactile sensation or sound.

[0019] Preferably, two sample adding holes are arranged in the upper cover, and a guiding structure is arranged in the sample adding holes, so that the sample can be added to the reaction cavity or to a separate sample cavity in two different ways, and the sample adding tube can only reach the cavity position where the sample needs to be added, avoiding the sample dripping into other holes.

[0020] In summary, the beneficial effects of the present invention are:

[0021] The assembled cartridge of the present invention can solve the limitations of conventional cartridges in terms of reagent compatibility, and the sources of these limitations include:

[0022] 1) Using microvalves or microfluidic channels to control the flow of liquids, which cannot adapt to and be compatible with liquid media with different properties (such as viscosity, surface tension, etc.). Microvalves and microfluidic channels will also cause varying degrees of liquid residue, which will also affect the detection performance;

[0023] 2) The shape of the reaction tube is different from that of the commonly used PCR tube, resulting in differences in the temperature change and temperature field of the reaction system; the fluorescence signals are also different;

[0024] 3) Different mixing methods also lead to differences in the reaction effect, such as the efficiency of magnetic bead nucleic acid capture and the differences in magnetic bead washing performance, etc.;

[0025] 4) The lack of a hot lid function also leads to differences in the temperature field of the reaction system during the reaction process, thus causing changes in the reaction effect;

[0026] By sealing the reaction detection area in the cartridge, the present invention can ensure that the environmental conditions of the reaction detection area are not contaminated by external air during the use of the cartridge, avoiding the impact on the detection accuracy. Moreover, during the heating process, it can also effectively avoid the temperature difference between the reaction well tube part and the top cover, so that the environmental conditions for reaction detection reach the optimal state;

[0027] At the same time, the tip of the pipette and its inner cavity are set to be non-coaxial, thereby effectively reducing the outer diameter size of the cartridge. During the movement of the pipette, maintaining the relative position accuracy with each cavity is a key factor affecting the sample addition performance, drainage residue, etc. The cooperation between the pipette and the hole pins of the upper cover can ensure that the position of the pipette is fixed when the cartridge is connected to the instrument, thus avoiding the position deviation of the pipette and each cavity caused by the connection process;

[0028] The present invention can detect samples of different methodologies by replacing different reaction detection tubes, such as: it can support colorimetric biochemical detection, can support immunoassay, can support real-time fluorescence quantitative PCR detection, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions in the embodiments of the invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the following drawings are only some embodiments of the invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0030] Figure 1Schematic diagram of the overall structure of a cartridge device for closed - type sample processing and detection with pipetting function according to the present invention;

[0031] Figure 2 Schematic diagram of the overall front - sectional view structure of a cartridge device for closed - type sample processing and detection with pipetting function according to the present invention;

[0032] Figure 3 Schematic diagram of the front - sectional view structure of a cartridge device for closed - type sample processing and detection with pipetting function according to the present invention;

[0033] Figure 4 Schematic diagram of the base structure of a cartridge device for closed - type sample processing and detection with pipetting function according to the present invention;

[0034] Figure 5 Schematic diagram of the top - view structure of the base of a cartridge device for closed - type sample processing and detection with pipetting function according to the present invention;

[0035] Figure 6 Schematic diagram of the structure at the sealing film of a cartridge device for closed - type sample processing and detection with pipetting function according to the present invention;

[0036] Figure 7 Schematic diagram of the tube cap structure of a cartridge device for closed - type sample processing and detection with pipetting function according to the present invention;

[0037] Figure 8 Schematic diagram of the reagent encapsulation in nucleic acid extraction of a cartridge device for closed - type sample processing and detection with pipetting function according to the present invention;

[0038] Figure 9 Schematic diagram of the nucleic acid extraction process of a cartridge device for closed - type sample processing and detection with pipetting function according to the present invention;

[0039] Figure 10 Schematic diagram of the main - view structure when the upper cover and the base of a cartridge device for closed - type sample processing and detection with pipetting function are opened according to the present invention;

[0040] Figure 11 Schematic diagram of the unfolded structure of the base and the reaction detection tube of a cartridge device for closed - type sample processing and detection with pipetting function according to the present invention;

[0041] Figure 12 Schematic diagram of the elastic bayonet structure at the upper cover and the base of a cartridge device for closed - type sample processing and detection with pipetting function according to the present invention.

[0042] The reference signs in the drawings are described separately as follows: 1. Base; 2. Upper cover; 3. Pipette; 4. Piston; 5. First sealing ring; 6. Second sealing ring; 7. Reaction detection tube; 8. Tube cap; 9. Sealing film; 10. Pressing plate; 11. Positioning hole; 12. Positioning pin; 13. Guide structure. Detailed implementation mode

[0043] All features disclosed in this specification, or all steps in the disclosed methods or processes, except for mutually exclusive features and / or steps, can be combined in any way.

[0044] Any feature disclosed in this specification (including any additional claims, abstract, and drawings), unless specifically stated, can be replaced by other equivalent or similar-purpose alternative features. That is, unless specifically stated, each feature is only an example in a series of equivalent or similar features.

[0045] In the present invention, unless otherwise clearly defined and limited, terms such as "installed", "connected", "connected", "fixed", etc. should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or integrated; it can be a mechanical connection, a direct connection, or an indirect connection through an intermediate medium, and it can be the communication inside at least two elements or the interaction relationship between at least two elements, unless otherwise clearly defined. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0046] The following is combined with Figures 1-12 The present invention will be described in detail. For the convenience of narration, the directions mentioned below are defined as follows: The up-down, left-right, front-back directions mentioned below are the same as the front-back, left-right, up-down directions of the Figure 1 viewing direction, Figure 1 and the directions shown are the same as the front-back, left-right, up-down directions of the front view direction of the device of the present invention.

[0047] Please refer to Figures 1-12, Embodiment 1 provided by the present invention: A cartridge device for closed sample processing and detection with pipetting function, including a base 1 and an upper cover 2. The upper cover 2 is rotatably installed on the base 1 to form a closed cavity, which can be disassembled. When operating, first inject the sample, and then connect the two to form a sealed space without additionally adding a sample injection port and a sealing structure for the sample injection port. There are indication marks on the upper cover 2 and the base 1 to facilitate aligning and rotating the positions of the two during disassembly and assembly. There are elastic bayonets on the upper cover 2 and the base 1. When the two are aligned, there is an obvious touch or sound. The base 1 is distributed with eleven chambers around the rotation center for holding samples, reagents, waste liquids, etc., or for reaction and detection. Among them, there is a main chamber in the middle of all chambers. At the area opposite to the main chamber in the base 1, four reaction and detection tubes 7 are provided, which can be an integral structure with the base 1 or separable, and are connected to the base 1 through interference fit, bonding, welding or clamping structure. There is a sleeve structure at a position deviating from the center in the upper cover 2 for installing a pipette 3 to form a closed cavity with the base 1. A second sealing ring 5 is provided at the connection between the upper cover 2 and the base 1. A first sealing ring 6 is installed between the pipette 3 and the side wall of the sleeve structure, and both are "O" - shaped sealing rings. The structure is simple, with low cost and good sealing effect. At the same time, a sealing structure for sealing the reaction and detection tube 7 is provided in the upper cover;

[0048] Among them, the sealing structure includes a guiding hole and a sealing cover provided in the upper cover. The guiding hole corresponds to the reaction and detection tube 7. In the free state, the sealing cover does not contact the base, but under an external action, the sealing cover can form a closed chamber with the reaction and detection tube 7. The sealing cover can be a sealing film 9 with elastic characteristics. Refer to Figure 6 , fixedly connected to the bottom of the upper cover 2 at the area where the guiding hole is located. The material can be rubber, silica gel, PP and other materials, and is transparent, so that the fluorescence signal of the reaction hole can be detected from above. Thus, under the action of an external instrument, using the heating rod on the pressing plate 10, insert the heating rod into the guiding hole, and the pressing plate 10 moves downward, so that the heating rod presses the sealing film 9 downward into the orifice position of the reaction and detection tube 7 for heating, thereby realizing sealing, and effectively avoiding the influence of the external environment during reaction detection;

[0049] The sealing cover can also be a tube cover 8 with a cylindrical structure and is transparent. Refer to Figure 7, the upper end of the tube cap 8 is concave in the middle, and the tube cap 8 can slide up and down in the guiding hole. The concave in the middle can just let the heating rod on the pressing plate 10 insert into it, so as to heat the tube cap 8 and seal the corresponding reaction detection tube 7. In this way, when heating and reacting the substances in the reaction detection tube 7, the heating rod on the top pressing plate 10 can also be used to abut against the tube cap 8 for heating. In this way, the temperatures of the bottom and the orifice part of the whole reaction detection tube 7 can be ensured to be controlled within the same range, so as to effectively avoid the generation of water droplets at the top of the reaction detection tube 7 when heating the reaction detection tube 7, which may contaminate the test substance, and the detection effect accuracy can be better;

[0050] The pipette 3 has an inner cavity, a tip and an anti-blocking structure. The tip and the inner cavity of the pipette are not coaxial, which can effectively reduce the outer diameter size of the cartridge. The eccentricity distance is ≥3 mm. The tip can puncture the sealing film 9 and reduce the liquid hanging on the outer wall, so as to reduce the contamination between reagents and improve the liquid injection accuracy;

[0051] The pipette 3 has hydrophobic and low adsorption characteristics, and the material is a polymer. There is a structure between the pipette 3 and the upper cover to prevent rotation and circumferential positioning. There is a positioning hole 11 at the upper end of the pipette 3, and a positioning pin 12 at the corresponding position on the upper cover 2, and the two achieve precise circumferential positioning of the pipette;

[0052] The bottom of the tip has an anti-blocking structure, which can be a wedge-shaped structure, or there are cross or cross-shaped grooves on the bottom plane;

[0053] A piston 4 is slidably arranged in the inner cavity of the pipette 3. The diameter of the opening on the pipette 3 is smaller than the diameter of the sealing part of the piston 4 to prevent the piston 4 from sliding out. There is a connection groove connected to the outside in the middle area of the upper end surface of the piston 4. Adding a piston inside helps to reduce the dead volume in the pipette and improve the pipetting accuracy;

[0054] There is a sample addition hole on the upper cover 2. In order to avoid accidentally adding samples to other holes during the sample addition operation or avoiding samples dripping onto other holes, there is a guiding structure 13 on the upper cover 2, so that the pipette can only reach the position of the sample addition cavity.

[0055] I. When performing pipetting operations using the cartridge:

[0056] ① Rotate the upper cover 2 relative to the base 1 so that the pipette 3 is aligned with the reaction detection tube 7 to be operated;

[0057] ② Lower the pipette 3 below the liquid level;

[0058] ③ Drive the external connection mechanism to move the piston 4 upward by a certain distance to complete liquid suction;

[0059] ④ Raise the liquid pipe 3 above the liquid level;

[0060] ⑤ Rotate the upper cover 2 relative to the base 1 to align the pipette 3 with another reaction detection tube 7 to be operated;

[0061] ⑥ Lower the pipette 3 to the bottom of the well;

[0062] ⑦ Drive the external connection mechanism to move the piston 4 downward by a certain distance to complete liquid discharge;

[0063] ⑧ Raise the pipette 3 above the base to complete pipetting.

[0064] II. When using the cartridge for mixing operations:

[0065] ① Rotate the upper cover 2 relative to the base 1 to align the pipette 3 with the reaction detection tube 7 to be operated;

[0066] ② Lower the pipette 3 below the liquid level;

[0067] ③ Drive the external connection mechanism to move the piston 4 upward by a certain distance to complete liquid suction;

[0068] ④ Reset the pipette 3 and rotate the upper cover 2 relative to the base 1 again to align the pipette 3 with another reaction detection tube 7 to be operated;

[0069] ⑤ Lower the pipette 3 by a certain distance;

[0070] ⑥ Drive the external connection mechanism to move the piston 4 upward by a certain distance to complete liquid suction;

[0071] ⑦ Reset the pipette 3 and rotate the upper cover 2 relative to the base 1 again to align the pipette 3 with an empty reaction detection tube 7;

[0072] ⑧ Lower the pipette 3 by a certain distance, drive the external connection mechanism to move the piston 4 downward by a certain distance to complete liquid discharge, so as to mix the liquids in the two reaction detection tubes 7 and discharge them into another reaction detection tube 7.

[0073] III. Nucleic acid extraction using the cartridge

[0074] The cartridge of the present invention can realize sample detection including different methodologies such as molecular, biochemical, and immunological. Taking nucleic acid detection (magnetic bead method nucleic acid extraction and real-time fluorescence quantitative PCR) as an example, the working process of the cartridge is referred to Figure 8

[0075] The reagents pre-packaged in the chamber of the base 1 and the reaction detection tube 7 are as follows:

[0076] 101: Lysis solution

[0077] 102: Magnetic beads

[0078] 112: Proteinase K

[0079] 103: Wash Solution I

[0080] 104: Wash Solution II

[0081] 105: Elution Solution

[0082] 106: Liquid Paraffin

[0083] 108: PCR Mix

[0084] 109: Internal Standard (IC)

[0085] 107-1: PCR Reaction Solution 1

[0086] 107-2: PCR Reaction Solution 2

[0087] 107-3: PCR Reaction Solution 3

[0088] 107-4: PCR Reaction Solution 4

[0089] 111: Waste Liquid Well

[0090] The workflow is as follows:

[0091] The first step

[0092] Manually add the sample to be analyzed into well 101, and then transfer magnetic beads (102), proteinase K (112) and IC (109) into well 101;

[0093] The second step

[0094] Perform lysis: Mix the liquid in reaction well 101, heat the liquid in reaction well 101 to a certain temperature, and the external magnetic field adsorbs the magnetic beads in well 101 to the side wall, and transfer the waste liquid in well 101 to waste liquid well 111;

[0095] The third step

[0096] Primary wash: Transfer wash solution I (103) into well 101, mix the liquid in reaction well 101, the external magnetic field adsorbs the magnetic beads in well 101 to the side wall, and transfer the waste liquid in well 101 to waste liquid well 111;

[0097] The fourth step

[0098] Secondary wash: Transfer wash solution II (104) into well 101, mix the liquid in reaction well 101, the external magnetic field adsorbs the magnetic beads in well 101 to the side wall, and transfer the waste liquid in well 101 to waste liquid well 111;

[0099] The fifth step

[0100] Elution: Transfer the eluent (105) to well 101, mix and heat the liquid in reaction well 101. The external magnetic field adsorbs the magnetic beads in well 101 to the side wall, and the remaining liquid in well 101 is the template to be amplified after extraction.

[0101] The sixth step

[0102] PCR system construction: Transfer the PCR mixture (108) and the template to be amplified (101) to wells 107-1 to 107-4, mix the liquid in wells 107-1 to 107-4, and transfer the liquid paraffin (106) to wells 107-1 to 107-4.

[0103] The seventh step

[0104] Amplification detection: Control the temperature of the liquid in reaction wells 107-1 to 107-4 to complete the amplification of the nucleic acid sequence, and detect the fluorescence signal in the reaction wells. According to the change of the fluorescence signal, calculate the concentration of nucleic acid in the reaction wells.

[0105] The detection of the entire nucleic acid is completed under sealed conditions, and the operation is simple and convenient, which can effectively ensure the accuracy of nucleic acid detection. Moreover, during the heating process, it can also effectively avoid the temperature difference between the tube part and the top cover of the reaction well, so that the environmental conditions for reaction detection reach the optimal state.

[0106] As described above, it is only the specific implementation manner of the invention, but the protection scope of the invention is not limited thereto. Any change or replacement that can be thought of without creative work should be covered within the protection scope of the invention. Therefore, the protection scope of the invention should be subject to the protection scope defined by the claims.

Claims

1. A cartridge device for closed sample processing and detection with pipetting function, comprising a base (1) and an upper cover (2), characterized in that: The upper cover (2) is rotatably mounted on the base (1). At least one chamber is distributed around the center in the base (1). Reaction detection tubes (7) are detachably arranged around the center at the bottom of the base (1). The upper cover (2) is provided with a sleeve structure at a position deviating from the center. A pipette (3) is slidably mounted up and down in the sleeve structure. A sealing structure for sealing the reaction detection tube (7) is provided in the upper cover (2).

2. The cartridge device for closed sample processing and detection with pipetting function according to claim 1, characterized in that: The sealing structure includes a guiding hole and a sealing cover provided in the upper cover (2). The guiding hole corresponds to the reaction detection tube (7).

3. The cartridge device for closed sample processing and detection with pipetting function according to claim 2, characterized in that: The sealing cover is an elastic sealing film (9), which is fixedly connected to the bottom of the upper cover (2) at the area where the guiding hole is located.

4. A cartridge device for closed sample processing and detection with a pipetting function according to claim 2, characterized in that: The sealing cover can also be a tube cover (8) with a cylindrical structure, and the tube cover (8) can slide up and down in the guiding hole.

5. A cartridge device for closed sample processing and detection with pipetting function according to claim 1, characterized in that: The tip at the bottom of the pipette (3) is not on the same straight line as the center line of the inner cavity on the upper side, and the eccentricity distance is ≥ 3 mm.

6. The cartridge device for closed sample processing and detection with pipetting function according to claim 5, characterized in that: The tip at the bottom of the pipette (3) has an anti-blocking structure. The anti-blocking structure is a wedge-shaped structure, or there are cross or cross-shaped grooves on the bottom plane. A positioning hole (11) is provided at the upper end of the pipette (3), and a positioning pin (12) is provided at the corresponding position on the upper cover (2).

7. A cartridge device for closed sample processing and detection with pipetting function according to claim 6, characterized in that: A piston (4) is slidably mounted in the inner cavity of the pipette (3). A connecting groove is provided in the middle of the upper end surface of the piston (4). The piston (4) is in sealing contact with the side wall of the inner cavity.

8. A cartridge device for closed sample processing and detection with pipetting function according to claim 7, characterized in that: A first sealing ring (6) is installed between the pipette (3) and the side wall of the sleeve structure. A second sealing ring (5) is installed between the outer end surface of the base (1) and the cavity wall at the bottom of the upper cover (2).

9. A cartridge device for closed sample processing and detection with a pipetting function according to claim 2, characterized in that: The sealing cover is made of a transparent material.

10. A cartridge device for closed sample processing and detection with pipetting function according to claims 1-9, characterized in that: The usage method includes the following steps: S1: Add the required reagents into each chamber of the base, and check whether film sealing is required according to the needs. S2: Install and connect the upper cover and the base to ensure the sealing of the connection. S3: Add the corresponding sample through the guiding structure in the sample addition port. S4: Cooperate with the instrument to automatically complete the pretreatment and reaction detection of the sample.

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