Reaction equipment

Abstract

The utility model relates to a PCR (polymerase chain reaction) technology, in particular to reaction equipment, which comprises a reaction tube, a reaction tube, a first electrode, a second electrode, a third electrode, a fourth electrode and a fifth electrode, the reaction tube comprises a tube body, and the tube body is provided with an opening and a containing cavity; and a movable block capable of moving is arranged in the accommodating cavity. According to the application, the active block moves in the limited thread, so that the reaction efficiency of PCR is effectively improved.

Description

Technical Field

[0001] The present application relates to the technical field of convection PCR (Polymerase Chain Reaction) reaction, and in particular to a reaction device. Background Art

[0002] In the field of biotechnology, polymerase chain reaction (PCR) has become a mature technology for amplifying specific nucleic acid sequences. This reaction process involves three main steps: denaturation, annealing, and extension, each of which requires a different reaction temperature. The temperature of denaturation is typically between 90 and 97°C; the temperature of annealing is selected based on the melting temperature of the primers used, typically between 50 and 65°C; the typical temperature of the extension reaction is 72°C.

[0003] Among them, the thermal convection polymerase chain reaction method mainly gives a higher temperature (such as above 95°C) to the bottom of the test tube containing the PCR mixture, and the rest of the test tube is exposed to room temperature air to dissipate heat, so that the temperature gradient from the bottom to the surface of the PCR mixture gradually decreases, such as from 95°C to 35°C, to induce thermal convection, so that the PCR mixture passes through different temperature sections during the convection process and performs different reaction steps. At present, thermal convection PCR is free convection PCR (Free Convective Polymerase Chain Reaction), that is, a technology that uses the principle of natural convection to achieve temperature cycling in PCR reactions.

[0004] In addition, CN101003825A discloses a method and device for nucleic acid sequence amplification using thermal convection. The amplification device includes: a plurality of heat sources that can provide heat to or remove heat from a plurality of specific areas of a sample contained in a reaction vessel, wherein the heat sources are designed to maintain a specific spatial temperature distribution of the sample, so that the higher temperature area is lower in height than the lower temperature area, wherein the specific spatial temperature distribution includes specific temperature areas that each implement a temperature condition, and wherein the specific spatial temperature distribution is a temperature distribution that causes sample circulation by thermal convection, whereby denaturation, annealing, and polymerization steps occur sequentially and repeatedly in the sample.

[0005] However, this traditional multi-heat source distributed amplification device design is mainly based on the heat convection of the reaction liquid at different temperatures, and the efficiency of the heat convection is low.

[0006] For example, Miao, Guijun et al. attempted to change the efficiency of free convection by inducing thermal convection in the capillary in "Free Convective PCR: From Principle Study to Commercial Applications—A Critical Review." (Analytica Chimica Acta 1108 (2020)).

[0007] For another example, patent application CN104611222A discloses a multi-stage thermal convection device and its use, which comprises a plurality of heat sources with different distribution positions, and uses the temperature difference of different heat sources to pass through corresponding temperature environments for different stages such as denaturation, annealing and polymerization steps.

[0008] In addition, patent application CN109401948A also discloses a one-stage thermal convection PCR instrument, in which the aluminum alloy block heating plate at the bottom of the instrument is used for heating, the temperature at the bottom of the capillary is controlled at the denaturation temperature, and the top of the capillary is at the annealing temperature. Under the effect of gravity difference and temperature difference, the reagents inside the capillary can continuously reach different temperatures through natural convection, thereby realizing PCR.

[0009] However, the above-mentioned traditional temperature control schemes all rely on the density difference of the reaction liquid for heat convection, and the efficiency of heat convection is very low. Summary of the invention

[0010] The purpose of the present application is to provide a PCR reaction device to partially solve or alleviate the above-mentioned deficiencies in the prior art, and to improve the reaction efficiency of PCR through an active convection mode; moreover, the active convection PCR in the present application may not rely on the effect of gravity and may be used in a zero-gravity environment, such as in a weightless space.

[0011] In order to solve the above-mentioned technical problems, the present application specifically adopts the following technical solutions: The first aspect of the present application is to provide a reaction device, comprising:

[0012] A reaction tube, the reaction tube comprising: a tube body, the tube body having an opening, and a receiving cavity; a first stopper is arranged inside the receiving cavity, and the first stopper divides the receiving cavity into a first receiving cavity and a second receiving cavity, the second receiving cavity is used to receive at least one movable block, and the diameter of the movable block is larger than the inner diameter of the stopper opening on the first stopper;

[0013] A magnetic driving module is arranged corresponding to one side of the reaction tube and is used to provide a corresponding magnetic force to the movable block, thereby guiding the movable block to reciprocate along the axial direction of the accommodating cavity on the axis of the second accommodating cavity under the action of the corresponding magnetic force, thereby driving the liquid in the accommodating cavity to circulate back and forth.

[0014] In some embodiments, the inner diameter of the first accommodating cavity is smaller than the inner diameter of the second accommodating cavity, the first limiter is a transition zone formed between the second end of the first accommodating cavity and the first end of the second accommodating cavity, and the inner diameter of the transition zone gradually increases in the direction from the first accommodating cavity to the second accommodating cavity.

[0015] In some embodiments, including:

[0016] A first temperature measuring module is disposed at a first end of the first accommodating cavity.

[0017] In some embodiments, including:

[0018] A second temperature measuring module, wherein the second temperature measuring module is arranged at the second end of the second containing cavity.

[0019] In some embodiments, a second limiting member is further provided in the second accommodating cavity, wherein the second limiting member includes: a limiting protrusion provided on the inner side of the second accommodating cavity, and a second limiting opening is formed on the inner edge of the limiting protrusion, and the inner diameter of the second limiting opening is smaller than the diameter of the movable block.

[0020] In some embodiments, the second limiting member includes: at least one limiting protrusion, and at least one connecting member for connecting the limiting protrusion and the tube wall of the second accommodating cavity; wherein, a plurality of the connecting members can be connected in a circle to form a columnar frame; the diameter of the columnar frame is greater than or equal to the inner diameter of the second accommodating cavity, and when the second limiting member is inserted into the inner side of the second accommodating cavity, the second limiting member will be slightly deformed under the extrusion of the second accommodating cavity, thereby enabling the second limiting member to be fixedly installed on the inner side of the second accommodating cavity.

[0021] In some embodiments, an inverted structure is provided at the first end of the connector, and a groove is provided in the inverted structure. When the tube wall of the second accommodating cavity is inserted into the inner side of the groove, the connector is fixed to the inner side of the second accommodating cavity.

[0022] In some embodiments, the inner diameter of the opening gradually decreases from the first end to the second end thereof, and the first end of the opening is a liquid inlet. In some embodiments, the movable block includes: at least one group of first protrusions arranged along a first direction, and at least one group of second protrusions arranged along a second direction, and the first direction intersects with the second direction, and the diameter of the movable block along the first direction and the diameter of the movable block along the second direction are both greater than the inner diameter of the limit opening.

[0023] In some embodiments, the movable mass is a permanent magnet. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings required for use in the embodiments or the prior art descriptions are briefly introduced below. In all the drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, each element or part is not necessarily drawn according to the actual scale. Obviously, the drawings described below are some embodiments of the present application. For those of ordinary skill in the art, other drawings can also be obtained based on these drawings without paying creative labor.

[0025] Figure 1 This is a schematic diagram of the reaction tube structure in an exemplary embodiment of the present application;

[0026] Figure 2 for Figure 1 A schematic diagram of the cross-sectional structure of the reaction tube shown;

[0027] Figure 3 This is a schematic diagram of the structure of a reaction tube in another embodiment of the present application;

[0028] Figure 4 for Figure 1 A schematic diagram of the internal structure of the reaction tube;

[0029] Figure 5 It is a schematic diagram of the structure of different activity blocks;

[0030] Figure 6 This is a schematic diagram of an active convection state in another exemplary embodiment of the present application;

[0031] Figure 7 This is a schematic diagram of the cross-sectional structure of a reaction tube in yet another embodiment of the present application;

[0032] Figure 8 This is a schematic diagram of a hollow structure of a movable block in an exemplary embodiment of the present application;

[0033] Fig. 9 This is a schematic diagram of the hollowed-out structure of a movable block in yet another exemplary embodiment of the present application.

[0034] Summary of reference numerals:

[0035] 10 is a reaction tube, 11 is an opening, 12 is a accommodating chamber, 121 is a first accommodating chamber, 122 is a second accommodating chamber, 13 is a first limiting member (also known as a transition zone), 14 is a second limiting member, 14a is a connecting member, and 14b is a limiting protrusion; 20 is a movable block, 21a is a first protrusion A, 21b is a first protrusion B, 22a is a second protrusion A, 22b is a second protrusion B, 30 is a heating module, L1 is a first axis, L2 is a second axis, I is a first temperature zone, and II is a second temperature zone. DETAILED DESCRIPTION

[0036] In order to make the purpose, technical scheme and advantages of the embodiments of the present application clearer, the technical scheme in the embodiments of the present application will be clearly and completely described below in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of this application.

[0037] Herein, suffixes such as "module", "component" or "unit" used to represent elements are only used to facilitate the description of the present application, and have no specific meanings by themselves. Therefore, "module", "component" or "unit" can be used mixedly.

[0038] In this document, the terms "upper", "lower", "inner", "outer", "front", "back", "one end", "the other end" and the like indicate positions or positional relationships based on the positions or positional relationships shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operate in a specific orientation, and therefore cannot be understood as limiting the present application. In addition, the terms "first" and "second" are used for descriptive purposes only, and cannot be understood as indicating or implying relative importance.

[0039] In this document, unless otherwise clearly specified and limited, the terms "installed", "provided with", "connected", etc. should be understood in a broad sense. For example, "connected" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection, a direct connection, or an indirect connection 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 this application can be understood according to specific circumstances.

[0040] Herein "and / or" includes any and all combinations of one or more of the associated listed items.

[0041] Herein, "plurality" means two or more than two, ie, it includes two, three, four, five, etc.

[0042] It should be noted that, in this article, the terms "include", "comprises" or any other variations thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, an element defined by the sentence "comprises a ..." does not exclude the existence of other identical elements in the process, method, article or device including the element.

[0043] As used in this specification, the term "about" typically means + / - 5% of the stated value, more typically + / - 4% of the stated value, more typically + / - 3% of the stated value, more typically + / - 2% of the stated value, even more typically + / - 1% of the stated value, and even more typically + / - 0.5% of the stated value.

[0044] In this specification, some embodiments may be disclosed in a format of being in a range. It should be understood that this description of "being in a range" is only for convenience and brevity, and should not be interpreted as a rigid limitation on the disclosed range. Therefore, the description of the range should be considered to have specifically disclosed all possible sub-ranges and independent numerical values ​​within this range. For example, the description of the range 1-6 should be considered to have specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., as well as individual numbers within this range, such as 1, 2, 3, 4, 5 and 6. Regardless of the breadth of the range, the above rules apply.

[0045] Herein, a "quasi-T-shaped cavity" (also referred to as a T-shaped cavity) refers to a reaction tube that can be placed in a horizontal state or a nearly horizontal state (such as tilted), and the opening of the reaction tube is set on the side wall of the tube body. Among them, the first axis L1 of the tube body of the reaction tube and the second axis L2 of the opening (set on the side wall of the tube body) intersect to form a quasi-T-shaped structure, so it is called a "quasi-T-shaped cavity".

[0046] Different from the traditional technical route of realizing free convection of liquid based on gravity and temperature difference, the present application proposes a technical route of realizing active convection of liquid.

[0047] Unlike the traditional free convection route, the active convection scheme in this application not only gets rid of the reliance on gravity in the liquid convection process, but also gets rid of the reliance on multi-point heat source distribution. That is, even with only a single heat source, this application can successfully complete the stable temperature distribution of PCR in the tube.

[0048] See also Figure 1-Figure 7 As shown, the present application provides a reaction device capable of active convection. Different from the traditional free convection PCR scheme, the active convection reaction device in the present application can get rid of the limitations of low liquid convection efficiency in free convection PCR and the dependence of the convection process on gravity. Therefore, the present application can be further applied to PCR reaction experiments in a weightless environment in space (such as a space station), such as using the PCR reaction device of the present application to quickly complete nucleic acid detection in a space station.

[0049] Embodiment 1

[0050] This application provides a PCR reaction device suitable for active convection scenarios, see Figure 5 As shown, the reaction equipment includes:

[0051] The reaction tube 10 comprises: a tube body having an opening 11 and a receiving cavity 12; a first stopper 13 is arranged inside the receiving cavity, and the first stopper is oriented toward a first axis L1 of the receiving cavity (the first axis L1 is as shown in FIG. Figure 2 The accommodating cavity 12 is formed with a first accommodating cavity 121 and a second accommodating cavity 122 on both sides of the first limiting piece, and at least one movable block 20 is arranged in the second accommodating cavity, and the diameter of the movable block is larger than the diameter of the limiting opening, so that the movable block is limited by the first limiting piece to move in the second accommodating cavity 122;

[0052] Preferably, in some embodiments, the diameter of the projection figure of the at least one movable block on the action surface during operation (such as reciprocating motion in the tube) is 30%-90% of the diameter of the accommodating cavity (such as the second accommodating cavity), and the action surface is a plane perpendicular or approximately perpendicular to the first axis;

[0053] In other words, the area of ​​the projection pattern of at least one movable block on the action surface during operation is 30%-90% of the cross-sectional area of ​​the accommodating cavity.

[0054] The active surface may refer to the plane where the cross-sectional direction of the accommodating cavity is located; the projection figure refers to the projection area formed by the at least one movable block on the active surface along the first axis direction.

[0055] Preferably, the diameter of the cross section of the movable block is 30%-90% of the diameter of the second accommodating cavity 122, so that the movable block can play a sufficiently large drainage role for the liquid in the accommodating cavity;

[0056] A magnetic driving module, which is arranged corresponding to one side of the reaction tube and is used to provide a corresponding magnetic force to the movable block 20, so as to guide the movable block to reciprocate along the axial direction of the second accommodating chamber 122 within a set distance under the action of the corresponding magnetic force, so as to drive convection between the liquid in the second accommodating chamber 122 and the liquid in the first accommodating chamber 121, thereby reducing the temperature difference of the liquid between the first accommodating chamber and the second accommodating chamber through active convection;

[0057] At least one heating module is disposed on one side of the reaction tube.

[0058] Preferably, in some embodiments, Figure 6 As shown, only one heating module 30 may be provided, and the heating module is provided at the first end of the first accommodating cavity 121 (ie, the end away from the second accommodating cavity 122 ).

[0059] In some embodiments, the inner diameter of the first accommodating cavity is smaller than the inner diameter of the second accommodating cavity, and the first limiting member 13 is a transition zone formed between the second end of the first accommodating cavity and the first end of the second accommodating cavity, and the inner diameter of the transition zone gradually increases along the direction from the first accommodating cavity to the second accommodating cavity, and the end of the transition zone close to the first accommodating cavity is the limiting opening, and the inner diameter of the limiting opening is smaller than the diameter of the movable block.

[0060] It should be noted that the active convection scheme proposed in the present application can also be applied to the reaction requirements of extreme PCR. Different from the reaction stages of traditional PCR, extreme PCR can limit the three reaction stages of PCR to two reaction stages, that is, the sample only needs to alternate between two temperature ranges to complete a cycle. For example, patent application CN104662160A discloses an extreme PCR, in which each cycle can be completed in less than 2 seconds, and even a single step such as annealing temperature only needs to be maintained for 0.05s-0.9s. However, the existing heat source setting method is difficult to meet the rapid cycle requirements under extreme PCR. On the contrary, the active convection scheme proposed in the present application can effectively ensure temperature stability on the basis of improving the efficiency of temperature cycling, thereby contributing to the normal reaction of extreme PCR.

[0061] See Figure 6As shown, the heating module 30 is arranged at the end of the first accommodating chamber 121 with a smaller tube diameter. During the heating process, the first accommodating chamber 121 and the second accommodating chamber 122 of the reaction tube will form a first temperature zone I and a second temperature zone II respectively, and the temperature of the first temperature zone I is higher than that of the second temperature zone II. When the sample completes a reaction in the first temperature zone I and the second temperature zone II in sequence, it is considered that a PCR cycle is completed.

[0062] Specifically, when the movable block reciprocates in the second accommodating chamber 122, the liquid with a lower temperature in the second accommodating chamber 122 will be driven to move toward the first accommodating chamber 121. Correspondingly, the liquid with a higher temperature in the first accommodating chamber 121 will also move toward the second accommodating chamber 122, thereby realizing active convection of the liquid under the two chambers (the liquid convection is shown in FIG. Figure 6 shown).

[0063] Alternatively, in other embodiments, the first limiting member may also be at least one limiting plate formed by a protrusion of the inner wall of the accommodating cavity, and the edge of at least one limiting plate cooperates with the inner wall of the accommodating cavity in the cross-sectional direction of the accommodating cavity (i.e., the cross-sectional direction in a direction perpendicular to the first axis L1) to form a limiting opening to allow liquid to flow between the first accommodating cavity and the second accommodating cavity, while limiting the movable block from moving from the second accommodating cavity to the first accommodating cavity.

[0064] In some embodiments, the opening is arranged on the side wall of the reaction chamber, and the second axis L2 of the opening intersects with the first axis L1, so that the opening and the reaction chamber together form a T-shaped cavity. Therefore, the reaction tube in this embodiment is also referred to as a T-tube. Preferably, the T-tube is suitable for a horizontally arranged PCR reaction scenario, in which case the liquid has no gravity effect in the horizontal direction, and thus it is difficult to achieve liquid convection using gravity.

[0065] Among them, since the T-shaped cavity has the characteristic of being placed horizontally, for batch PCR reactions, the purpose of high-throughput detection can be achieved by stacking multiple layers. For example, in some embodiments, the same heating module can be arranged in the vertical direction, and multiple reaction tubes of the T-shaped cavity can be stacked in the vertical direction to form a reaction tube array suitable for high-throughput detection.

[0066] And because it is placed horizontally, the device can be made flat to flexibly meet the adaptation requirements of different detection scenarios or supporting detection equipment.

[0067] In some embodiments, the inner diameter of the opening gradually decreases from the first end to the second end, and the first end of the opening is a liquid inlet. Preferably, the opening is configured to be smaller at the top and larger at the bottom to prevent liquid from splashing out.

[0068] In some embodiments, Figure 2 As shown, the movable block can be spherical, or the movable block can be cylindrical. For example, when the movable block is cylindrical, its two ends are designed to be spherical (such as hemispherical) to keep its corners in a curved surface design.

[0069] In some embodiments, the movable block 20 includes: at least one group of first protrusions (including: first protrusion A21a, first protrusion B21b) arranged along the first direction, and at least one group of second protrusions (including second protrusion A22a, second protrusion B22b) arranged along the second direction, and the first direction intersects with the second direction, and the diameter of the movable block along the first direction and the diameter of the movable block along the second direction are both greater than the inner diameter of the limit opening. Wherein, a spacing area is formed between adjacent protrusions for liquid circulation.

[0070] In this embodiment, the multi-protrusion movable block design can not only cooperate with the limiting opening to play a limiting role, but also the spacing area between the protrusions can help provide sufficient movement space for liquid convection.

[0071] Preferably, the edge of the cross section of the movable block is composed of multiple smooth curves to prevent the movable block from damaging the inner wall of the reaction tube when reciprocating.

[0072] In some embodiments, the heating module is disposed at the first end of the first accommodating cavity 121 .

[0073] In some embodiments, two or more heating modules may also be set. For example, one heating module may be set at the bottom of the PCR tube (such as the first end of the first accommodating cavity) and one heating module may be set at the middle of the PCR tube (such as between the first and second accommodating cavities).

[0074] In some embodiments, including:

[0075] A first temperature measuring module, wherein the first temperature measuring module is arranged at a first end of the first containing cavity;

[0076] The second temperature measuring module is arranged at the second end of the second accommodating cavity. That is to say, in this embodiment, two temperature measuring points are arranged at the heat source and the far heat source respectively, so as to monitor the temperature difference between the two ends in real time.

[0077] In some embodiments, the magnetic force driving module may be a Hall effect sensor. The Hall effect sensor can sense and determine the magnetic field direction of the movable block (such as a permanent magnet), thereby applying a reverse magnetic field force to form a driving force for the movable block to move toward the second end of the second accommodating cavity to move the movable block, and stop the reverse magnetic field force according to the distance of the magnetic field. At this time, if the reaction tube is placed at an angle, the movable block slowly falls to the bottom of the tube due to gravity.

[0078] It can be understood that the size of the PCR reaction tube in the present application can be designed using the size of a conventional PCR tube.

[0079] Furthermore, the active convection solution proposed in the present application avoids the limitations of conducting convection PCR reactions in capillaries, which can save application costs while improving convection efficiency.

[0080] It should be understood that there are at least the following limitations for conducting PCR reactions in capillaries: 1) The viscosity of the reaction liquid will seriously affect the convection efficiency; 2) The diameter of the capillary is related to the liquid level. When the liquid level is not appropriate, such as when insufficient or excessive amounts of liquid are added, the efficiency of natural convection will be seriously affected; 3) Specific capillary consumables are required, which lack a certain degree of versatility and have a relatively high application cost.

[0081] In some embodiments, the movable block can be placed in the PCR reaction tube during the processing stage. Alternatively, in other embodiments, the movable block can also be manually placed in the reaction tube by a staff member before the reaction (e.g. Figure 2 (shown in the middle right figure).

[0082] In some embodiments, the number of active blocks may be set to one.

[0083] For example, in some embodiments, the movable block may be a solid structure, such as a solid magnet.

[0084] For example, in some embodiments, the movable block may also be a hollow structure (also known as a grid structure or a mesh structure). Figure 8 As shown, the movable block may be provided with a plurality of mesh holes penetrating the movable block.

[0085] For another example, in some embodiments, the movable block may be a three-dimensional mesh structure. Specifically, the movable block may have a spherical surface (or the surface may also be set to other geometric shapes), and the spherical surface is provided with a plurality of mesh holes penetrating the surface.

[0086] Alternatively, in other embodiments, the number of movable blocks can be set to two or more. For example, in some embodiments, a plurality of small-diameter magnetic particles (such as magnet beads) can be set inside the chamber, wherein the plurality of magnetic particles can be focused together to form a large magnet bead, or a grid-like structure. The area of ​​the projection pattern of this large magnet bead or the formed grid-like structure on the action surface can be 30%-90% of the cross-sectional area of ​​the accommodating cavity.

[0087] See also Fig. 9As shown, in some embodiments, the movable block is a magnetic particle, and a plurality of magnetic particles are arranged in the receiving chamber (preferably the second receiving chamber); wherein the plurality of magnetic particles are connected to each other under the action of a corresponding magnetic force, and pores are formed between the contact points or contact surfaces between the adjacent plurality of magnetic particles, so that the plurality of magnetic particles form a mesh structure. wherein, when the mesh structure moves back and forth in the tube driven by magnetic force, liquid can pass through the pores.

[0088] In some embodiments, the structural design of the PCR reaction tube (also known as a T-type tube) that can be used horizontally can be seen in Figure 1-Figure 5 As shown, the structure of the magnetic element can also be seen Fig. 9 shown.

[0089] It should be noted that the T-shaped tube can be placed in a horizontal or approximately horizontal state in actual application, or it can be placed at a certain tilt angle (the tilt angle refers to the angle between the first axis of the T-shaped tube and the horizontal plane). For example, the T-shaped tube can be placed at an angle of about 0-30° with the horizontal plane during use, or it can be placed at an angle of about 0-45° with the horizontal plane. Of course, it is understandable that the specific tilt angle can be adaptively adjusted by the user in combination with the supporting equipment, and this application does not limit it.

[0090] Embodiment 2

[0091] The present application also proposes a PCR reaction tube suitable for use in a gravity scenario. Unlike the reaction tube in Example 1, the reaction tube in the present application can be placed in a vertical or nearly vertical state (for example, the reaction tube has a certain inclination angle with the vertical direction) during the reaction process (therefore, it can also be called a vertical tube), and unlike the T-tube, the opening of the reaction tube in this embodiment is set at the second end of the second reaction chamber.

[0092] like Figure 7 As shown, the reaction tube in the embodiment of the present application includes: an opening 11 arranged at the end of the reaction tube 10, and a movable block (not shown in the figure) is arranged inside the reaction tube. Preferably, a transition zone 13 (equivalent to the first stopper) is arranged inside the reaction tube 10, and the transition zone 13 divides the reaction tube 10 into a first accommodating chamber 121 and a second accommodating chamber 122; wherein, the inner diameter of the transition zone 13 gradually decreases from the first end (i.e., the side connected to the second accommodating chamber 122) to the second end; and the second end of the second accommodating chamber 122 is formed with an opening 11 for adding samples.

[0093] Furthermore, in some embodiments, a second limiting member 14 is also provided in the second accommodating cavity 122, wherein the second limiting member 14 includes: a limiting protrusion provided on the inner side of the second accommodating cavity 122, and a second limiting opening is formed on the edge of the limiting protrusion facing inward (i.e., the side facing the first axis L1 direction), and an inner diameter of the second limiting opening is smaller than the diameter of the movable block, so as to further limit the movement of the movable block within the second accommodating cavity 122.

[0094] For example, in some embodiments, the second limiting member may be configured as a detachable structure, such as the second limiting member 14 includes: at least one limiting protrusion 14b, and at least one connecting member 14a for connecting the limiting protrusion 14b and the tube wall.

[0095] Specifically, a plurality of connecting members 14a are connected in a circumferential manner to form a column frame, the diameter of which is equal to or slightly larger than the inner diameter of the second accommodating cavity 122, and when the second stopper 14 is inserted into the inner side of the second accommodating cavity 122, the second stopper 14 will be slightly deformed under the pressure of the second accommodating cavity 122, so that the second stopper 14 can be fixedly installed inside the second accommodating cavity 122. Alternatively, the first end of the connecting member 14a (the second end of which is connected to the stopper protrusion) can be provided with an undercut structure, and the undercut structure has a groove, and when the tube wall of the second accommodating cavity 122 is inserted into the inner side of the groove, it can also be fixed inside the second accommodating cavity 122.

[0096] It can be understood that the reaction tube or reaction equipment in this embodiment may include the same or similar structures or components as those in the first embodiment, which will not be described in detail here.

[0097] In some embodiments, the structural design of the vertical tube suitable for vertical placement can be seen in Figure 6-Figure 8 shown.

[0098] It can be understood that in actual application, the vertical tube in this embodiment can be placed at a vertical or approximately vertical angle, or it can also be placed at a certain inclination angle with the vertical direction (wherein the inclination angle refers to the angle between the first axis and the vertical direction / vertical plane).

[0099] For example, during use, the vertical tube can be placed at an angle of about 0-30° to the vertical direction, or can be placed at an angle of about 0-45° to the vertical direction. Of course, it is understood that the specific tilt angle can be adaptively adjusted by the user in combination with supporting equipment, and this application does not limit it.

[0100] Embodiment 3

[0101] The present application further provides a control method for an active convection reaction device, comprising the steps of:

[0102] S101 provides a reaction device, the reaction device comprising: a tube body, the tube body having an opening 11, and a first accommodating cavity 121 and a second accommodating cavity 122 formed in the tube body, a limiting opening formed between the first accommodating cavity 121 and the second accommodating cavity 122, for limiting the movable block located in the second accommodating cavity from moving into the first accommodating cavity; a magnetic driving device disposed outside the reaction tube, the magnetic driving device for providing a magnetic force to the movable block;

[0103] It can be understood that the reaction equipment described in any of the above embodiments can be used in this embodiment.

[0104] S102: detecting and obtaining a first temperature at a first end of the first accommodating chamber, and detecting and obtaining a second temperature at a second end of the second accommodating chamber;

[0105] S103, when it is detected that a first temperature difference between the first temperature and the second temperature is greater than a set temperature difference threshold, controlling the movable block to move forward or backward along the first axis direction of the reaction chamber at a specific speed mode through a magnetic driving device;

[0106] S104: After the first time, calculate the second temperature difference between the current second temperature and the preset target temperature, and adjust the specific speed mode according to the second temperature difference; wherein the adjustment step includes:

[0107] When the second temperature difference is less than the first temperature difference threshold, accelerating the specific speed mode;

[0108] When the second temperature difference is greater than or equal to the first temperature difference threshold and less than the second temperature difference threshold, maintaining the specific speed mode;

[0109] When the second temperature difference is greater than or equal to the second temperature difference threshold and less than a third temperature difference threshold, the specific speed mode is decelerated.

[0110] It is worth noting that in the existing natural thermal convection PCR process, due to the low convection efficiency, the PCR amplification efficiency is also low, so the sensitivity of temperature difference detection is not high. On the contrary, the active convection mode proposed in this application not only improves the convection efficiency, but also improves the PCR amplification efficiency, thereby improving the temperature detection sensitivity.

[0111] Furthermore, in this embodiment, active convection and temperature difference monitoring are used in coordination to dynamically and in real time regulate a specific speed mode, and the stability of the temperature difference distribution can be effectively maintained through rapid micro-regulation.

[0112] It can be understood that the specific values ​​of the set temperature difference threshold, temperature difference threshold, target temperature, etc. in this embodiment can be adaptively set according to the actual PCR conditions.

[0113] In particular, in some embodiments, when the second temperature difference is small (eg, when the second temperature is greater than the target temperature), the specific speed mode may be directly reduced to zero.

[0114] In some embodiments, the reaction device further includes: a cooling module disposed at the second end of the second reaction chamber. Correspondingly, the method further includes the steps of:

[0115] When the second temperature difference is greater than the third temperature difference, the temperature of the liquid at the second end of the second reaction chamber is reduced by the temperature reduction module.

[0116] For example, in some embodiments, the cooling module may be a refrigeration plate, or the cooling module may also have a pipeline of a low-temperature medium.

[0117] For example, in some embodiments, at least one heating module can be provided at each end of the reaction tube, and a cooling module can be added if necessary (equivalent to providing different heat transfer sources on the periphery of the reaction tube). In some specific embodiments, one of the heat transfer sources can complete high-temperature heating (such as a temperature ≥ 90°C), while the temperature of one of the heat transfer sources is lower than about 40°C (or even 0°C, forming a high temperature at one end and a low temperature at the other end, to achieve rapid heating and cooling of the liquid).

[0118] In this embodiment, a specific speed mode is used in combination with the actual temperature difference to perform graded speed adjustment on the movable block, so that the movement mode (ie, movement speed) of the movable block can be quickly adjusted according to different temperature difference sizes.

[0119] Furthermore, for the active convection scheme based on the movable block, the present application also proposes a restrictive movement mode, and preferably sets the active thread of the movable block on the side away from the heat source, which is beneficial for maintaining a relatively stable temperature difference environment for the PCR reaction (avoiding excessive convection) to form relatively stable multi-stage temperature ranges; at the same time, the mutual coordination of the restrictive movement mode and the multi-stage adjustment scheme can effectively avoid or reduce the generation of bubbles or aerosols due to the movement of the movable block.

[0120] Furthermore, in some embodiments, the method further comprises:

[0121] When the movable block moves, detecting the bubble content in the liquid;

[0122] When the bubble content is higher than a set bubble content, the specific speed mode is decelerated.

[0123] In this embodiment, the movable block is adjusted in time through the bubble detection feedback result, which can effectively control the amount of bubble generation to prevent the activity of the movable block from interfering with the accuracy and reliability of the PCR reaction.

[0124] In some embodiments, the specific speed mode in S103 is determined by the following steps:

[0125] When the first temperature difference belongs to a first temperature difference range, a first specific speed mode is adopted;

[0126] When the first temperature difference belongs to a second temperature difference range, a second specific speed mode is adopted; wherein the first temperature difference range is smaller than the second temperature difference range, and correspondingly the specific speed in the first specific speed mode is smaller than the specific speed in the second specific speed mode.

[0127] The movable block can have different movement speeds by adjusting the magnitude of the magnetic force.

[0128] It is understandable that in a PCR reaction scenario, the reaction time of the sample in a specific temperature range is very short. Once a certain area in the tube fails to reach the specified target temperature within the set time, it is very easy to cause incomplete reaction during the PCR cycle.

[0129] In this regard, in this embodiment, a simple interval adjustment method is directly adopted in the initial state, which can effectively simplify the speed control method of the movable block, improve the response efficiency of the movable block, and avoid excessive adjustment time.

[0130] In addition, the present application further uses specific parameters (i.e., the second temperature difference) to flexibly and hierarchically adjust the specific speed mode selected in the initial state, so as to quickly respond to the temperature difference change and adjust the active convection state within a limited time. Thus, the temperature at both ends of the PCR tube can be maintained within a specific temperature range, while avoiding the generation of a large number of bubbles during a long adjustment process to interfere with the normal reaction of PCR.

[0131] It should be noted that, in this article, the terms "include", "comprises" or any other variations thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, an element defined by the sentence "comprises a ..." does not exclude the existence of other identical elements in the process, method, article or device including the element.

[0132] Through the description of the above implementation methods, those skilled in the art can clearly understand that the above-mentioned embodiment methods can be implemented by means of software plus a necessary general hardware platform, and of course by hardware, but in many cases the former is a better implementation method. Based on such an understanding, the technical solution of the present application, or the part that contributes to the prior art, can be embodied in the form of a software product, which is stored in a storage medium (such as ROM / RAM, a disk, or an optical disk), and includes a number of instructions for a computer terminal (which can be a mobile phone, a computer, a server, or a network device, etc.) to execute the methods described in each embodiment of the present application.

[0133] The embodiments of the present application are described above in conjunction with the accompanying drawings, but the present application is not limited to the above-mentioned specific implementation methods. The above-mentioned specific implementation methods are merely illustrative and not restrictive. Under the guidance of the present application, ordinary technicians in this field can also make many forms without departing from the purpose of the present application and the scope of protection of the claims, all of which are within the protection of the present application.

Claims

1. A reaction device, characterized in that: include: A reaction tube (10), the reaction tube (10) comprising: a tube body, the tube body having an opening (11), and a receiving cavity (12); a first stopper is arranged inside the receiving cavity (12), and the first stopper divides the receiving cavity into a first receiving cavity (121) and a second receiving cavity (122); the second receiving cavity (12) is used to receive at least one movable block (20), and the diameter of the movable block (20) is greater than the inner diameter of the stopper opening on the first stopper; A magnetic driving module is arranged corresponding to one side of the reaction tube and is used to provide a corresponding magnetic force to the movable block (20), thereby guiding the movable block to reciprocate along the axial direction of the accommodating chamber (12) on the axis of the second accommodating chamber (122) under the action of the corresponding magnetic force, thereby driving the liquid in the accommodating chamber (12) to circulate back and forth.

2. The reaction device according to claim 1, characterized in that: The inner diameter of the first accommodating cavity is smaller than the inner diameter of the second accommodating cavity. The first limiter is a transition zone formed between the second end of the first accommodating cavity and the first end of the second accommodating cavity. The inner diameter of the transition zone gradually increases along the direction from the first accommodating cavity to the second accommodating cavity.

3. The reaction device according to claim 1, characterized in that: include: A first temperature measuring module is disposed at a first end of the first accommodating cavity.

4. The reaction device according to claim 3, characterized in that: include: A second temperature measuring module, wherein the second temperature measuring module is arranged at the second end of the second containing cavity.

5. The reaction device according to claim 1, characterized in that: A second limiting member (14) is also provided in the second accommodating cavity (122), wherein the second limiting member (14) comprises: a limiting protrusion provided on the inner side of the second accommodating cavity (122), and a second limiting opening is formed on the inner edge of the limiting protrusion, and the inner diameter of the second limiting opening is smaller than the diameter of the movable block (20).

6. The reaction device according to claim 5, characterized in that: The second limiting member (14) comprises: at least one limiting protrusion (14b), and at least one connecting member (14a) for connecting the limiting protrusion (14b) and the tube wall of the second accommodating cavity (122); wherein a plurality of the connecting members (14a) can be connected in a circumferential manner to form a columnar frame; the diameter of the columnar frame is greater than or equal to the inner diameter of the second accommodating cavity (122), and when the second limiting member (14) is inserted into the inner side of the second accommodating cavity (122), the second limiting member (14) will be slightly deformed under the pressure of the second accommodating cavity (122), thereby enabling the second limiting member (14) to be fixedly installed inside the second accommodating cavity (122).

7. The reaction device according to claim 6, characterized in that: The first end of the connecting piece (14a) is provided with an undercut structure, and a groove is provided inside the undercut structure. When the tube wall of the second accommodating cavity (122) is inserted into the inner side of the groove, the connecting piece (14a) is fixed inside the second accommodating cavity (122).

8. The reaction device according to claim 1, characterized in that: The inner diameter of the opening gradually decreases from the first end to the second end thereof, and the first end of the opening is a liquid inlet.

9. The reaction device according to claim 1, characterized in that: The movable block (20) comprises: at least one group of first protrusions arranged along a first direction, and at least one group of second protrusions arranged along a second direction, and the first direction intersects with the second direction, and the diameter of the movable block along the first direction and the diameter of the movable block along the second direction are both greater than the inner diameter of the limit opening.

10. The reaction device according to claim 6, characterized in that: The movable block is a permanent magnet.

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