Multi-channel nucleic acid molecule detection device

Abstract

The utility model discloses a multi-channel nucleic acid molecule detection device which comprises a detection box shell, a liquid inlet is arranged on the detection box shell, a liquid separation cavity, a negative pressure cavity, a plurality of buffer cavities and a plurality of amplification cavities are arranged in the detection box shell, the liquid separation cavity is communicated with the liquid inlet, amplification reagents are arranged in the amplification cavities, and the negative pressure cavity is communicated with the negative pressure cavity. The plurality of buffer cavities are communicated with the liquid separation cavity and are communicated with inlets of the plurality of amplification cavities in a one-to-one correspondence manner, the negative pressure cavity is communicated with outlets of the plurality of amplification cavities, and thermally unstable sealing films are arranged between the negative pressure cavity and the amplification cavities in a sealing manner; the cracking container is suitable for being mounted on the detection box shell and is communicated with the liquid inlet after being mounted, a sample adding opening is formed in the cracking container, and a cracking reagent is arranged in the cracking container. According to the multi-channel nucleic acid molecule detection device disclosed by the embodiment of the utility model, amplification of a plurality of target sequences can be simultaneously carried out, quantitative sample adding can be realized, and the multi-channel nucleic acid molecule detection device has the advantages of accuracy in detection, strong applicability, convenience in operation and the like.

Description

Technical Field

[0001] The utility model relates to the technical field of nucleic acid detection, and in particular to a multi-channel nucleic acid molecule detection device. Background Art

[0002] Constant temperature amplification technology can be combined with portable equipment and test kits to achieve nucleic acid amplification outside the laboratory, such as outdoor field environments, epidemic prevention and control sites, etc., and has important application value. However, compared with polymerase chain reaction technology, it has certain limitations when amplifying multiple target sequences.

[0003] Portable nucleic acid detection devices that use constant temperature amplification technology in related technologies have difficulty in performing quantitative control of samples and reagents when simultaneously amplifying multiple target sequences, resulting in inconsistent amounts of samples and reagents allocated to each channel, affecting the accuracy of the detection results. Utility Model Content

[0004] The utility model aims to solve at least one of the technical problems existing in the prior art. To this end, the utility model proposes a multi-channel nucleic acid molecule detection device, which can simultaneously amplify multiple target sequences, can achieve quantitative sampling, and has the advantages of accurate detection, strong applicability, and convenient operation.

[0005] To achieve the above-mentioned purpose, according to an embodiment of the utility model, a multi-channel nucleic acid molecule detection device is proposed, and the multi-channel nucleic acid molecule detection device includes: a detection box shell, a liquid inlet is provided on the detection box shell, a liquid separation chamber, a negative pressure chamber, a plurality of buffer chambers and a plurality of amplification chambers are provided in the detection box shell, the liquid separation chamber is connected with the liquid inlet, an amplification reagent is provided in the amplification chamber, the plurality of buffer chambers are connected with the liquid separation chamber and are connected with the inlets of the plurality of amplification chambers one by one, the negative pressure chamber is respectively connected with the outlets of the plurality of amplification chambers, and a thermally unstable sealing film is sealed between the negative pressure chamber and the amplification chamber; a lysis container, the lysis container is suitable for being installed on the detection box shell and is connected with the liquid inlet after installation, a sample addition port is provided on the lysis container, and a lysis reagent is provided in the lysis container.

[0006] The multi-channel nucleic acid molecule detection device according to the embodiment of the utility model can amplify multiple target sequences at the same time, can realize quantitative sampling, and has the advantages of accurate detection, strong applicability, and convenient operation.

[0007] In addition, the multi-channel nucleic acid molecule detection device according to the above embodiment of the utility model may also have the following additional technical features:

[0008] According to an embodiment of the present invention, the buffer cavity is formed on the inner bottom surface of the liquid separation cavity, the amplification cavity is located below the buffer cavity, and the cross-sectional area of ​​the buffer cavity gradually decreases from top to bottom.

[0009] According to an embodiment of the present invention, the buffer chamber is connected to the amplification chamber via a connecting flow channel, and the connecting flow channel has a wavy bending section.

[0010] According to an embodiment of the present invention, the inner bottom surface of the liquid separation chamber is oriented in a horizontal direction.

[0011] According to an embodiment of the present invention, the volume of each buffer chamber is 20-50 microliters.

[0012] According to one embodiment of the utility model, the sample addition port is arranged on the upper surface of the lysis container and can be opened and closed, the lower surface of the lysis container is provided with a puncturable sealing film, the upper surface of the detection box shell is provided with a connecting card slot, the lysis container is suitable for being loaded into the connecting card slot, the inner bottom surface of the connecting card slot is provided with a hollow puncture piece, when the lysis container is loaded into the connecting card slot, the hollow puncture piece passes through the puncturable sealing film, and the hollow puncture piece is connected to the liquid inlet.

[0013] According to an embodiment of the present utility model, a waste liquid chamber is further provided in the detection box housing, and the waste liquid chamber is formed on the inner bottom surface of the liquid separation chamber.

[0014] According to an embodiment of the utility model, the outlet of each amplification chamber is provided with a waterproof and breathable membrane.

[0015] According to an embodiment of the present utility model, a transparent observation window suitable for observing the amplification chamber is provided on the detection box housing.

[0016] According to an embodiment of the utility model, the negative pressure chamber has a port, and the port is connected to the plurality of amplification chambers respectively through gas channels, and the thermally unstable sealing film seals the port.

[0017] Additional aspects and advantages of the present invention will be given in part in the following description, and in part will become apparent from the following description, or will be learned through the practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The above and / or additional aspects and advantages of the present invention will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

[0019] Figure 1 It is a schematic structural diagram of a multi-channel nucleic acid molecule detection device according to an embodiment of the utility model.

[0020] Figure numerals: multi-channel nucleic acid molecule detection device 1, liquid separation chamber 11, negative pressure chamber 12, amplification chamber 13, buffer chamber 14, connecting flow channel 15, bending section 16, connecting card slot 17, waste liquid chamber 18, gas channel 19, lysis container 20, sample addition port 21, puncturable sealing membrane 22, thermally unstable sealing membrane 30, hollow piercing element 40, waterproof breathable membrane 50. DETAILED DESCRIPTION

[0021] The embodiments of the present invention are described in detail below, and examples of the embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and cannot be understood as limiting the present invention.

[0022] In the description of the present utility model, it is necessary to understand that 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" and the like indicate positions or positional relationships based on the positions or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present utility model and simplifying the description, rather than indicating or implying 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 utility model. In addition, features defined as "first" and "second" may explicitly or implicitly include one or more of such features. In the description of the present utility model, unless otherwise specified, "multiple" means two or more.

[0023] In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the terms "installation", "connection" and "connection" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0024] The multi-channel nucleic acid molecule detection device 1 according to an embodiment of the present utility model is described below with reference to the accompanying drawings.

[0025] like Figure 1 As shown, the multi-channel nucleic acid molecule detection device 1 according to an embodiment of the present utility model includes a detection box housing and a lysis container 20.

[0026] The detection box housing is provided with a liquid inlet, and the detection box housing is provided with a liquid separation chamber 11, a negative pressure chamber 12, a plurality of buffer chambers 14 and a plurality of amplification chambers 13. The liquid separation chamber 11 is connected to the liquid inlet, and an amplification reagent is provided in the amplification chamber 13. The plurality of buffer chambers 14 are all connected to the liquid separation chamber 11 and are connected to the inlets of the plurality of amplification chambers 13 one by one. The negative pressure chamber 12 is respectively connected to the outlets of the plurality of amplification chambers 13, and a thermally unstable sealing film is sealed between the negative pressure chamber 12 and the amplification chamber 13. The lysis container 20 is suitable for being installed on the detection box housing and is connected to the liquid inlet after installation. The lysis container 20 is provided with a sample addition port 21, and a lysis reagent is provided in the lysis container 20.

[0027] Specifically, the sample adding port 21 may be constructed as an openable and closable structure, which is opened when adding samples and closed after adding samples to prevent secondary contamination caused by aerosols.

[0028] During detection, the sample inlet 21 of the lysis container 20 is first opened, and a sample is added into the lysis container 20 through the sample inlet 21 and mixed with the lysis reagent to fully lyse the sample and release nucleic acid, and then the sample inlet 21 is closed.

[0029] The lysis container 20 is mounted on the detection box housing so that the lysis container 20 is connected to the liquid inlet, and the liquid in the lysis container 20 flows into the liquid separation chamber 11 through the liquid inlet.

[0030] The liquid passes through the liquid separation chamber 11 and fills the plurality of buffer chambers 14 in sequence.

[0031] The heat-unstable sealing film 30 sealing between the negative pressure chamber 12 and the amplification chamber 13 is heated and melted to connect the negative pressure chamber 12 with the amplification chamber 13. The gas in each amplification chamber 13 is sucked into the negative pressure chamber 12. At the same time, under the action of air pressure, the liquid buffered in each buffer chamber 14 enters the amplification chamber 13 and mixes with the amplification reagent pre-stored in the amplification chamber 13 to perform constant temperature amplification.

[0032] The amplification results are detected, such as fluorescence detection or color judgment.

[0033] Specifically, the amplification reagent may include one or more of an enzyme system, specific upstream and downstream primers, deoxyribonucleoside triphosphates (dNTPs), a buffer system magnesium ion, etc., and may also selectively include one or more of a specific fluorescent probe, a color indicator, and a fluorescent dye to achieve fluorescence detection. The specific upstream and downstream primers and specific probes in each amplification chamber 13 may be different, and are designed according to the corresponding detection targets to achieve simultaneous detection of different detection targets.

[0034] For example, in some embodiments, the amplification reagent includes an enzyme system, specific upstream and downstream primers, deoxyribonucleoside triphosphates, specific fluorescent probes, a buffer system, and magnesium ions. The enzyme system includes a recombinase that can bind to an oligonucleotide primer, a single-stranded DNA binding protein, a strand displacement DNA polymerase, and a nuclease exonuclease that cuts the fluorescent probe. The fluorescent probe contains a base analog tetrahydrofuran molecule, which has a fluorescent group and a quenching group on both sides of the tetrahydrofuran molecule, and a blocker that prevents the probe from extending at the 3 ends of the probe. The exonuclease, as a DNA repair enzyme, recognizes the tetrahydrofuran site and cuts the probe to separate the fluorescent group and the quenching group to generate fluorescence, so that fluorescence detection can be performed after amplification. In this embodiment, the heating temperature can be 30 degrees Celsius.

[0035] In other embodiments, the amplification reagent comprises an enzyme system, three groups of specific primers, deoxyribonucleoside triphosphates, a color indicator, a buffer system, and magnesium ions. The enzyme system comprises a recombinase that can bind to oligonucleotide primers, a single-stranded DNA binding protein, a strand displacement DNA polymerase, and an exonuclease that cuts the fluorescent probe. Three specific groups of primers (FIP / BIP, FIP / BIP, FIP / BIP) are designed according to the different target molecules to be tested in each amplification chamber. The color indicator can be one of L-HNB and phenol red, so that the test results can be judged by color after amplification. In this embodiment, the heating temperature can be 65 degrees Celsius and last for 20 minutes.

[0036] According to the multi-channel nucleic acid molecule detection device 1 of the embodiment of the utility model, by setting up multiple buffer chambers 14 and negative pressure chambers 12, the lysis reagent of the mixed sample in the lysis container 20 enters the liquid separation chamber 11 through the liquid inlet, and then enters the buffer chamber 14 first. After filling the multiple buffer chambers 14, the negative pressure chamber 12 is opened by heating, so that the reagent in the buffer chamber 14 is sucked into the amplification chamber 13 for amplification reaction. This can ensure that enough samples and reagents enter each amplification chamber 13, realize quantitative addition of reagents in each buffer chamber 14, facilitate multi-channel simultaneous amplification of multiple target sequences, and improve the accuracy of detection results when multiple target sequences are detected simultaneously.

[0037] Furthermore, since the heat-unstable sealing film 30 is melted by heating to generate power in the negative pressure chamber 12 to suck the reagent from the buffer chamber 14 into the amplification chamber 13, it can not only provide power for the flow of the reagent and ensure that the reagent completely enters the amplification chamber 13, thereby further ensuring the accuracy of the detection result, but also only requires a heating step to achieve the connection between the negative pressure chamber 12 and the amplification chamber 13, which is simple and convenient to operate. Since the constant temperature amplification technology itself also requires heating, there is no need for additional external devices to provide driving force for the reagent, which facilitates the operation of the multi-channel nucleic acid molecule detection device 1, reduces the restrictions of other external devices on the use occasions of the multi-channel nucleic acid molecule detection device 1, facilitates on-site detection of the multi-channel nucleic acid molecule detection device 1, and improves the applicability of the multi-channel nucleic acid molecule detection device 1.

[0038] Therefore, the multi-channel nucleic acid molecule detection device 1 according to the embodiment of the utility model can amplify multiple target sequences at the same time, can realize quantitative sampling, and has the advantages of accurate detection, strong applicability, and convenient operation.

[0039] The following describes a multi-channel nucleic acid molecule detection device 1 according to a specific embodiment of the present utility model with reference to the accompanying drawings.

[0040] In some specific embodiments of the present invention, Figure 1 As shown, the multi-channel nucleic acid molecule detection device 1 according to an embodiment of the present utility model includes a detection box housing and a lysis container 20.

[0041] Advantageously, if Figure 1 As shown, the buffer chamber 14 is formed on the inner bottom surface of the liquid separation chamber 11 (the up and down directions are shown by the arrows in the figure), the amplification chamber 13 is located below the buffer chamber 14, and the cross-sectional area of ​​the buffer chamber 14 gradually decreases from top to bottom. In this way, it is convenient for the liquid in the liquid separation chamber 11 to flow into the buffer chamber 14, so that the liquid fills the buffer chamber 14 so as to quantitatively control the pre-stored liquid to be entered into the amplification chamber 13.

[0042] More advantageously, if Figure 1 As shown, the buffer chamber 14 is connected to the amplification chamber 13 through a connecting channel 15, and the connecting channel 15 has a wavy bending section 16. This can prevent the liquid in the buffer chamber 14 from directly entering the amplification chamber 13, so that the liquid in the buffer chamber 14 flows into the amplification chamber 13 under pressure after the negative pressure chamber 12 is opened, so that the liquid fills the buffer chamber 14 before entering the amplification chamber 13, thereby further facilitating the quantitative control of the liquid.

[0043] Furthermore, if Figure 1 As shown, the inner bottom surface of the liquid separation chamber 11 is oriented in the horizontal direction, so that the liquid in the liquid separation chamber 11 can flow into the plurality of buffer chambers 14 evenly, and further facilitate quantitative control of the liquid.

[0044] Alternatively, if Figure 1 As shown, the volume of each buffer chamber 14 is 20-50 microliters. 25 microliters is preferred. This makes it easy to quantitatively control the reagents while ensuring that the amplification chamber 13 has enough reagents for the amplification reaction.

[0045] Specifically, Figure 1 As shown, the detection box housing is further provided with a waste liquid chamber 18, which is formed on the inner bottom surface of the liquid separation chamber 11. In this way, the excess reagent can enter the waste liquid chamber 18, so that the reagent in the lysis container 20 can be greater than the sum of the volumes of the multiple buffer chambers 14, ensuring that each buffer chamber 14 is allocated with sufficient reagent, and further facilitating quantitative control of the reagent.

[0046] Optionally, the number of buffer chambers 14 may be 3-8, and 3 is preferred.

[0047] Figure 1 FIG. 1 shows a multi-channel nucleic acid molecule detection device 1 according to some embodiments of the present utility model. Figure 1 As shown, the sample addition port 21 is provided on the upper surface of the lysis container 20 and can be opened and closed, the lower surface of the lysis container 20 is provided with a puncturable sealing film 22, the upper surface of the detection box housing is provided with a connecting card slot 17, the lysis container 20 is suitable for being loaded into the connecting card slot 17, and the inner bottom surface of the connecting card slot 17 is provided with a hollow puncture member 40. When the lysis container 20 is loaded into the connecting card slot 17, the hollow puncture member 40 passes through the puncturable sealing film 22, and the hollow puncture member 40 is connected with the liquid inlet. Specifically, the puncturable sealing film 22 can be an aluminum film. In this way, after the sample is added through the sample addition port 21, the lysis container 20 can be loaded into the connecting card slot 17. During the loading process, the hollow puncture member 40 punctures the puncturable sealing film 22 to connect the lysis container 20 and the liquid inlet, so that the liquid in the lysis container 20 enters the liquid separation chamber 11. In this way, after adding the sample, you only need to put the lysis container 20 into the connecting card slot 17 and heat it to automatically perform subsequent steps such as one-to-many, amplification, and detection without the need for other operations or secondary adding of samples, thereby simplifying the operating procedures of the multi-channel nucleic acid molecule detection device 1 and making the operation of the multi-channel nucleic acid molecule detection device 1 simpler and more convenient.

[0048] Specifically, Figure 1 As shown, the outlet of each amplification chamber 13 is provided with a waterproof breathable membrane 50. In this way, the gas in the amplification chamber 13 can be discharged through the waterproof breathable membrane 50 and enter the negative pressure chamber 12, while the liquid is blocked in the amplification chamber 13 by the waterproof breathable membrane 50, which further facilitates the quantitative control of the reagent in the amplification chamber 13 and prevents the liquid from blocking the gas path.

[0049] Optionally, the detection box housing is provided with a transparent observation window suitable for observing the amplification chamber 13. Specifically, the cross section of the amplification chamber 13 can be circular, and the two opposite surfaces of the amplification chamber 13 in the thickness direction of the detection box housing can be made of transparent material, so as to facilitate detection after amplification.

[0050] Specifically, Figure 1 As shown, the negative pressure chamber 12 has a port, which is connected to the multiple amplification chambers 13 through the gas channel 19, and the thermally unstable sealing film 30 seals the port. Specifically, the gas channel 19 may include a main path and multiple branches, the main path is connected to the port, and the multiple branches are respectively connected to the outlets of the multiple amplification chambers 13, and the thermally unstable sealing film 30 seals between the port and the main path. In this way, it is convenient to use the negative pressure chamber 12 to drive the flow of gas in the amplification chamber 13 and liquid in the buffer chamber 14.

[0051] Other structures and operations of the multi-channel nucleic acid molecule detection device 1 according to the embodiment of the present invention are known to ordinary technicians in the field and will not be described in detail here.

[0052] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "illustrative embodiments", "examples", "specific examples", or "some examples" means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the utility model. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0053] Although the embodiments of the present invention have been shown and described, those skilled in the art will appreciate that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the claims and their equivalents.

Claims

1. A multi-channel nucleic acid molecule detection device, characterized in that: include: A detection box shell, wherein the detection box shell is provided with a liquid inlet, and the detection box shell is provided with a liquid separation chamber, a negative pressure chamber, a plurality of buffer chambers and a plurality of amplification chambers, the liquid separation chamber is connected with the liquid inlet, an amplification reagent is provided in the amplification chamber, the plurality of buffer chambers are all connected with the liquid separation chamber and are connected with the inlets of the plurality of amplification chambers one by one, the negative pressure chamber is respectively connected with the outlets of the plurality of amplification chambers, and a thermally unstable sealing film is sealed between the negative pressure chamber and the amplification chamber; A lysis container, wherein the lysis container is suitable for being installed on the detection box housing and is connected with the liquid inlet after installation. The lysis container is provided with a sample addition port, and a lysis reagent is provided in the lysis container.

2. The multi-channel nucleic acid molecule detection device according to claim 1, characterized in that: The buffer cavity is formed on the inner bottom surface of the liquid separation cavity, the amplification cavity is located below the buffer cavity, and the cross-sectional area of ​​the buffer cavity gradually decreases from top to bottom.

3. The multi-channel nucleic acid molecule detection device according to claim 2, characterized in that: The buffer chamber is connected to the amplification chamber through a connecting flow channel, and the connecting flow channel has a wavy bending section.

4. The multi-channel nucleic acid molecule detection device according to claim 2, characterized in that: The inner bottom surface of the liquid separation chamber is oriented in a horizontal direction.

5. The multi-channel nucleic acid molecule detection device according to claim 1, characterized in that: The volume of each buffer chamber is 20-50 μl.

6. The multi-channel nucleic acid molecule detection device according to claim 1, characterized in that: The sample addition port is arranged on the upper surface of the lysis container and can be opened and closed. The lower surface of the lysis container is provided with a puncturable sealing film. The upper surface of the detection box shell is provided with a connecting slot. The lysis container is suitable for being loaded into the connecting slot. The inner bottom surface of the connecting slot is provided with a hollow puncture piece. When the lysis container is loaded into the connecting slot, the hollow puncture piece passes through the puncturable sealing film, and the hollow puncture piece is connected to the liquid inlet.

7. The multi-channel nucleic acid molecule detection device according to claim 1, characterized in that: A waste liquid cavity is also provided in the detection box housing, and the waste liquid cavity is formed on the inner bottom surface of the liquid separation cavity.

8. The multi-channel nucleic acid molecule detection device according to claim 1, characterized in that: The outlet of each amplification chamber is provided with a waterproof and breathable membrane.

9. The multi-channel nucleic acid molecule detection device according to claim 1, characterized in that: The detection box housing is provided with a transparent observation window suitable for observing the amplification chamber.

10. The multi-channel nucleic acid molecule detection device according to claim 1, characterized in that: The negative pressure chamber has a port, and the port is connected to the plurality of amplification chambers through gas channels, respectively, and the thermally unstable sealing film seals the port.

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