CN118165804A - Kit, amplification equipment and closed amplification detection method - Google Patents

Abstract

The invention discloses a kit, an amplifying device and a closed state amplifying detection method, which relate to the technical field of small amplifying devices, in particular to a kit related to portable amplifying devices; the first flow channel and the second flow channel can be effectively and reliably closed in a destructive manner simultaneously only by configuring an external function in a closed operation area, so that the high-cost scheme of configuring different types of microfluidic valves by multiple amplification cavities is abandoned, and the problem that leakage pollution risks exist in recovery treatment of the used reagent kits in consumable materials is solved.

Description

Kit, amplification equipment and closed amplification detection method

Technical Field

The invention relates to the technical field of small-sized amplification equipment, in particular to a kit related to portable amplification equipment, and specifically relates to a kit, amplification equipment and a closed amplification detection method.

Background

In biomedical, chemical and environmental testing, etc., separation of components of interest from a sample fluid is important for quantitative or qualitative analysis, and these target substances to be analyzed are usually in a low concentration distribution state in various samples, and it is very difficult to analyze them directly. The molecular diagnosis technology using the nucleic acid sequence as an analysis object is very important in medical quarantine scenes such as disease diagnosis, epidemic prevention and control, accurate medicine instruction and the like, and plays a very important role in scenes such as criminal investigation, agriculture, animal husbandry and the like. With the development of the detection system based on the molecular diagnosis technology, an effective technical support is provided for the development of the on-chip detection system for the instant diagnosis and integration in the medical detection, and meanwhile, the requirement of lower operation specialty is met.

In practical use, the implementation scheme of instant diagnosis is various, different companies design different types of integrated consumables to realize the integrated output from a sample to a result on a small amplification device, a plurality of disc type amplification consumables are used, the unique disc type structure of the consumables can use the centrifugal force of rotation as a driving force, chinese patent nos. 201710371949.6 and 201711091035.0 disclose a disc type consumable, the front side and the back side of the disc type consumable are provided with different processing cavities and flow channels, and different parts are provided with a plurality of valve members, the corresponding valve is opened as required and communicated with the exhaust part, the sample liquid or the corresponding treatment liquid is circulated or mixed by utilizing different rotation speeds and rotation directions, and the corresponding valve is required to be closed to complete amplification in the amplification process, so that the design requires the configuration of a plurality of valves, and the complex and accurate rotation and valve opening and closing control are required in different time periods, so that the realization difficulty is very high; in the scheme disclosed in China patent 200980162435.3, the detection consumable box can be designed into a sector, so that in rotary centrifugal driving equipment, a plurality of consumable boxes can be arranged in parallel, multiple detection of a plurality of samples can be realized, a plurality of bulges are arranged in the system, switching of an opening or closing state is realized through overlap joint and staggered angle change of the bulges, a plurality of groups of valves which are independently controlled are replaced to a certain extent, the structural arrangement needs to be matched by a specific system, the special system has no advantage in opening and closing of a runner with a tiny size, and the requirement on the processing precision of the bulges is very high, so that the high-precision driving and closing of the bulges are realized; of course, some schemes like those disclosed in chinese patent 201510889385.6 can utilize bending or necking to form capillary parts and other technologies to form valve-like members with preset resistance, so as to save the switch of the valve members, and the sealing resistance between the treatment chambers formed by the design is different, and the sealing reliability is low, especially for the temperature cycle type PCR amplification chamber, the effective sealing is difficult to realize, the probability of generating bubbles at high temperature is very high, and the amplification detection result is inaccurate or the result repeatability is poor; there are other types of non-centrifugal force driven kits, of course, european patent EP2018723956 discloses the use of a pierceable membrane to close different chambers and pre-disposition of various types of liquid or lyophilized reagents in the different chambers, however, this design is not applicable to an amplification chamber, and the closure of the amplification chamber cannot be achieved once the pierceable membrane is pierced; the scheme disclosed in European patent EP3458609B1 is a scheme for realizing consumable boxes and related drives adopted by Roche small-sized amplification equipment, the adopted consumable is soft consumable, liquid is driven to transfer between different chambers through a plurality of groups of extrusion units, the liquid after extraction is extruded into the amplification consumable connected with the bottom to perform amplification, the sealing of the amplification process is dependent on the extrusion force exerted by the extrusion units, and although the effect is better in sealing a larger amplification system and a large-size flow channel, the sealing is very difficult for the small-size flow channel below micrometers; in practice many scholars have studied the technical difficulties of cyclic amplification of small-volume amplification chambers with a volume of typically not more than 50 μl, and as a result, found that in microfluidic chips, during the execution of PCR (polymerase chain reaction) reactions, fluid evaporation and loss may occur due to the concentrated heat applied to the reaction area and high heat flux density, besides, there is a problem that fluid leaks from the microfluidic chip, and further, due to the high reaction temperature in the reaction area, there is a risk that a large amount of bubbles are generated by fluid evaporation, which causes not only difficulty in accurate measurement of the optical system in the reaction area, but also variation of the reaction system due to bubbles, thereby causing distortion of the results, the accuracy and reliability of the system are lower.

In order to ensure that the kit used by the small-sized amplification equipment can accurately perform amplification, a low-cost scheme which has simple sealing structure, single corresponding configuration structure and reliability is designed to solve the problem to be solved.

Disclosure of Invention

The invention aims at: in view of the above problems, the present invention provides a kit, an amplification apparatus, and a closed amplification detection method, which reliably ensure the pressure in an amplification chamber by destructively sealing a connecting channel, can prevent the generation of bubbles in an amplification system liquid during high-low temperature cyclic amplification, and can also enable a plurality of amplification chambers to be in independent closed states by destructively sealing operation, so that the problems of steam generation and mutual interference are hardly caused; the first flow channel and the second flow channel can be effectively and reliably closed in a destructive manner simultaneously only by configuring an external function in a closed operation area, so that the high-cost scheme of configuring different types of microfluidic valves by multiple amplification cavities is abandoned, and the problem that leakage pollution risks exist in recovery treatment of the used reagent kits in consumable materials is solved.

The technical scheme adopted by the invention is as follows:

The utility model provides a kit, includes the kit body, this internal system chamber that is equipped with of kit, amplification chamber, divide liquid drive portion and intercommunication runner, the intercommunication runner includes first runner and second runner, the one end of first runner communicates in the top of amplification chamber, the other end intercommunication divides liquid drive portion, the one end of second runner communicates in the middle part of amplification chamber, the other end intercommunication system chamber, divide liquid drive portion accessible pressure variation drive system intracavity wait to distribute liquid to fill in amplification chamber through the second runner, first runner and second runner can be simultaneously destructively sealed.

Further, a closing operation area for destructively closing the first flow channel and the second flow channel is further arranged in the kit body, a plurality of closing auxiliary parts are arranged in the closing operation area, and the closing auxiliary parts are arranged between the first flow channel and the second flow channel at intervals, so that the first flow channel and the second flow channel are separated at intervals.

Further, the communication flow channel comprises a parallel flow channel section matched with the length direction of the reagent box, the parallel flow channel section comprises a first flow channel parallel section corresponding to the first flow channel and a second flow channel parallel section corresponding to the second flow channel, and at least part of the first flow channel parallel section and the second flow channel parallel section are arranged in the closed operation area.

Further, a closed reinforcing part is arranged at the corresponding position of the parallel flow passage section and the closed operation area, and the flow passage sectional area of the position of the parallel flow passage section, where the closed reinforcing part is arranged, is smaller than the flow passage sectional area of the position, where the closed reinforcing part is not arranged. Preferably, the cross-sectional area of the flow passage where the closed reinforcement portion is arranged in the parallel flow passage segment is 1/3 to 4/5 of the cross-sectional area of the flow passage where the closed reinforcement portion is not arranged.

Further, the liquid separation driving part is provided with a deformation recovery piece, the deformation recovery piece can deform when being extruded by external force, and the suction force is recovered and generated when no external force acts, so that liquid to be distributed in the system cavity is filled in the amplification cavity through the second flow channel.

Further, the area above the center line of the amplification cavity is a configuration part with the cross-sectional area gradually decreasing from bottom to top.

Further, a buffer cavity is further arranged in the kit body, the buffer cavity is communicated with the system cavity through a buffer communication flow passage, and a flow control valve is arranged at the buffer communication flow passage; the buffer cavity is also provided with an openable connecting cover, the buffer cavity receives external liquid into the buffer cavity through the connecting cover, the kit body is also provided with a driving gas injection port and a vent port, the driving gas injection port is communicated with the buffer cavity and can blow driving gas flow into the buffer cavity, and the vent port is communicated with the system cavity and can supplement gas into the system cavity.

Further, the number of the amplification chambers is even, liquid separation driving units matched with the number of the amplification chambers are arranged in the liquid separation driving part, each amplification chamber is communicated with the corresponding liquid separation driving unit through the corresponding first flow channel and the common flow channel of the system chamber through the corresponding second flow channel, and the even amplification chambers are symmetrically distributed on two sides of the distribution flow channel section of the common flow channel.

An amplification apparatus comprising the kit, and further comprising

A kit receiving location capable of receiving and confining the kit;

the clamping device can apply clamping or releasing operation to the liquid separation driving part to enable the liquid separation driving part to generate pressure change and drive liquid to be distributed in the system cavity to be filled in the amplification cavity through the second flow channel;

A closure device capable of applying a destructive closure operation to the first flow path and the second flow path;

a heating module capable of performing a cyclic heating operation on the closed amplification chamber;

The detection module can cooperate with the heating module to perform fluorescence acquisition operation on the sealed amplification cavity in the cyclic heating process, so as to obtain an amplification detection result.

A closed amplification detection method is applied to the amplification equipment and comprises the following steps,

Filling: starting a liquid separation driving part to generate pressure difference between the amplification cavity and the system cavity, and driving liquid to be distributed in the system cavity to be filled in the amplification cavity to obtain a filled amplification cavity;

and (3) a sealing step: under the external action applied by the sealing device, the first flow channel and the second flow channel are simultaneously and destructively sealed, so that the filling-state amplification cavity is isolated and sealed, and the liquid in the filling-state amplification cavity is used as the amplification system liquid;

Amplification detection step: and (3) amplifying the amplification system liquid in the amplification cavity in a closed state, and performing fluorescence detection on the amplification system liquid in the amplification to obtain an amplification detection result.

In summary, due to the adoption of the technical scheme, the beneficial effects of the invention are as follows:

1. According to the invention, through the advantage analysis of the traditional closed-cover amplification scheme, the technical scheme of sealing the amplification chambers in the kit matched with the small-sized amplification equipment is obtained, and the pressure in the amplification chambers can be reliably ensured through the destructive sealing connecting flow channels, so that the low-pressure leakage state similar to a pressure cooker or even the non-pressure leakage state can be formed, the generation of bubbles in the amplification system liquid can be avoided in the high-low temperature cyclic amplification, and the destructive sealing operation also enables the amplification chambers to be in independent sealing states, so that the problems of steam generation and mutual interference are hardly caused.

2. The invention ensures that the first flow channel and the second flow channel comprise parallel flow channel sections, and at least part of the parallel flow channel sections can be arranged in the closed operation area, so that the first flow channel and the second flow channel can be effectively and reliably and simultaneously destructively sealed by only configuring external action in the closed operation area, the high-cost scheme of configuring different types of microfluidic valves by multiple amplification cavities is abandoned, and the problem of leakage pollution risk in recovery treatment of the reagent box which is completed in consumable materials is also solved.

3. The invention is provided with the public flow channel for a plurality of second flow channels to communicate with the system cavity, and even amplification cavities can be symmetrically arranged at two sides of the distribution flow channel section of the public flow channel.

4. The parallel runner sections in the closed operation area are internally provided with the closed reinforcing parts, so that the small and concentrated area of the broken runner required by the closing operation can be ensured, and the closed reinforcing parts are matched with a plurality of closed auxiliary parts which are distributed at intervals, so that the breaking resistance is reduced, the deformation of the broken closure can be accommodated, the internal stress is released, and the broken closure is ensured to be more reliable.

5. The plurality of liquid-dividing driving units corresponding to the number of the amplification chambers are independently configured so that the liquid in the system chambers can be more uniformly and uniformly sucked and distributed into not less than two amplification chambers.

6. The area above the central line of the amplification cavity comprises the configuration part with the cross-sectional area gradually reduced from bottom to top, and the connecting port of the first flow channel is arranged at the top of the amplification cavity, so that the turbulence degree in the filling process is smaller, the filling basically does not generate air gaps, and the filling degree is higher.

7. The configuration of the buffer cavity of the invention enables the application scene of the kit to be wider and enables the liquid in the system cavity to be more uniformly mixed.

8. The closed amplification detection method provided by the application utilizes the kit, and performs destructive closure of the connecting flow channel after the amplification cavity is filled to form an independent amplification reaction cavity, so that the pressure required in the amplification process can be ensured, the liquid in the system can be prevented from evaporating, and a thermal cover similar to a traditional system is not required to be arranged.

Drawings

FIG. 1 is a schematic diagram of the structure of a kit of the present invention;

FIG. 2 is a state diagram of the system within the cartridge containing the liquid to be dispensed;

FIG. 3 is a cross-sectional view in the direction A-A of FIG. 2 showing the liquid separation driving part in a state of being pressed and deformed;

FIG. 4 is a cross-sectional view taken along line A-A of FIG. 2 illustrating the liquid separation drive portion being released;

FIG. 5 is a cross-sectional view taken along the direction A-A of FIG. 2 illustrating the recovery of deformation of the liquid separation driving portion;

FIG. 6 is a diagram of a process of dispensing liquid into the system cavity corresponding to FIG. 4;

FIG. 7 is a diagram of a process of dispensing liquid into the system cavity corresponding to FIG. 5;

FIG. 8 is a front view of the closing of the connection flow passage with the completed filling in the closed operating area by external action;

FIG. 9 is a side view of the closure of the connection channel with the completed filling in the closed operating area by external action;

FIG. 10 is a schematic diagram showing the state of the first flow channel and the second flow channel of the amplification chamber before filling in accordance with the present invention;

FIG. 11 is a schematic diagram showing the state of the first flow channel and the second flow channel of the amplification chamber after filling.

The marks in the figure: 10-amplification chamber, 11-first flow channel, 111-first flow channel parallel section, 12-second flow channel, 121-second flow channel parallel section, 20-liquid separation driving part, 30-system chamber, 311-distribution flow channel section, 32-driving gas injection port, 40-buffer chamber, 41-flow control valve, 42-connection cover, 43-vent port, 100-closing operation area, 210-closing strengthening part, 211-closing auxiliary part, 500-closing device and 600-clamping device.

Detailed Description

The present invention will be described in detail with reference to the accompanying drawings.

The present invention will be described in further detail with reference to the drawings and examples, in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.

Example 1

The kit provided by the invention is configured into a basic rectangular shape, as shown in fig. 1, the length direction of the kit is shown from top to bottom in the figure, the direction perpendicular to the length direction is the width direction of the kit, at least two amplification chambers 10 are arranged at intervals in the middle area of the kit, the number of the amplification chambers 10 can be set arbitrarily according to the needs, the number of the amplification chambers 10 can be optimally configured into an even number in order to ensure the balance of flow resistance, the amplification chambers 10 are optimally configured into a shape with the cross-sectional area changing, and the shape comprises a liquid drop type or a liquid-like drop type, an egg type, an ellipsoid, a cone-like body or other revolution bodies, so that the sufficient extension in the liquid filling process can be ensured, the air gap probability is smaller due to smaller turbulence, the filling degree of the amplification cavity 10 is higher, the filling degree of 95% -100% can be realized, the subsequent amplification is more reliable, and the detection accuracy is higher. The device also comprises a connecting runner, wherein the connecting runner comprises a first runner 11, a second runner 12 and a public runner, and a connecting port of the first runner 11 is arranged at the top of the amplification cavity 10, so that the top area of the amplification cavity 10 can be filled at last, the possibility of gas residue is reduced, and the filling degree is ensured to be higher; in this embodiment, the area of the amplification chamber 10 near the top may specifically be the area above the center line of the amplification chamber 10, including the configuration portion with the cross-sectional area gradually decreasing from bottom to top, so that a certain compression constraint can be applied to the liquid to be filled later, and a rectification effect is achieved, and finally the filling degree of the amplification chamber 10 is basically close to 100%, and no gas remains basically. The dispensing liquid inlet of the amplification chamber 10, that is, the connection port of the second flow channel 12, is disposed near the center line of the amplification chamber 10, so that the dual effects of gravity and abrupt cross-section change can be utilized to enable the liquid filled into the amplification chamber 10 to be substantially adhered to the wall, thereby reducing the probability of turbulence and air gap generation, and if the connection port of the second flow channel 12 is disposed at the bottom of the amplification chamber 10, a high-turbulence filling state similar to a fountain is more likely to be generated, which is unfavorable for high-filling and filling without air gap. Before filling the amplification chamber 10, the first flow channel 11 can be in fluid communication with the liquid-dividing driving part 20, the second flow channel 12 can be in fluid communication with the system chamber 30, at least part of the parallel flow channel sections exist between the first flow channel 11 and the second flow channel 12, the first flow channel 11 is provided with first flow channel parallel sections 111 which are all arranged in parallel along the length direction of the reagent box, the second flow channel 12 is provided with second flow channel parallel sections 121 which are all arranged in parallel along the length direction of the reagent box, thus forming parallel flow channel sections which are arranged along the length direction of the reagent box and are twice as many as the amplification chamber 10, and the first flow channel 11 and the second flow channel 12 are at least partially arranged in the closed operation area 100, the amplification chamber 10 can be filled under the driving of the liquid-dividing driving part 20, and the first flow channel 11 and the second flow channel 12 in the closed operation region 100 can be simultaneously destructively closed under the external action applied by the closing means 500, so that not less than two of the filled amplification chambers 10 can amplify the amplification system liquid therein in a closed state, preferably, the first flow channel parallel section 111 and the second flow channel parallel section 121 corresponding to the closed operation region 100 here, the destructively closed under the external action here results from the first flow channel parallel section 111 and the second flow channel parallel section 121 in the closed operation region 100, the length of the closed operation region 100 is configured to be substantially identical to the width dimension of the kit, and may be particularly 0.9 to 1.1 times the width dimension of the kit to achieve that each parallel flow channel segment can be reliably closed without excessive waste of space and corresponding driving force, where the length of the closed operating region 100 may exceed the width of the kit because the closed operating region 100 is an external action contact location such as welding or fusing, which may result in the length of the closed operating region 100 exceeding the width of the kit.

In this embodiment, the liquid-separating driving portion 20 includes liquid-separating driving units corresponding to the amplifying chambers 10, and each liquid-separating driving unit includes a deformation recovery member, and forms a pumping action in the recovery process, so that the liquid to be dispensed in the system chamber 30 is injected into the amplifying chambers 10 through the second flow channel 12, where the deformation recovery member may be an independent element, including an air bag or an air film member configured by using a leather material, a rubber material, or the like, or may be a non-independent element, or may be a semi-closed chamber or a hollow portion on a substrate of the kit, and the deformation recovery film may be made of materials such as propylene, polyethylene, polystyrene, cyclic Olefin Copolymer (COC), polyester film, and polyacetate. In order to ensure that the temperature rising and falling operation can be performed quickly and efficiently by the amplification system liquid in each amplification cavity 10, the volume of each amplification cavity 10 is configured to be 5-50 mu L, more preferably the volume of each amplification cavity 10 is configured to be 20-40 mu L, the volume of each amplification cavity 10 can be configured to be 30 mu L under special conditions, thus being matched with a liquid-dividing driving unit with a preset deformation volume capacity, the low-resistance quick and uniform liquid distribution transfer operation can be realized, the filling-state amplification cavity 10 with the filling degree close to 100% is completed, the connecting flow channel can be destructively closed in a closed operation area 100, the amplification cavities 10 are further heated even by temperature circulation, the amplification cavities 10 are in a closed high-pressure amplification state because the amplification system liquid in the amplification cavities 10 does not shrink with temperature change, the interior of the amplification cavities 10 is also irrelevant to the air pressure of the external environment, the low-pressure leakage like high-pressure even the non-pressure leakage state can be formed in the interior of the amplification cavity, in such a low-temperature circulation state can be realized, the destruction reaction liquid can be generated, the multiple independent cavities 10 are also in a closed operation state, the same type of the self-contained state is almost no interference can be generated, the problem is solved, the problem of the recovery of the multiple-bubble-free air bubble can be solved, and the mutual pollution can be solved, and the problem is solved, and the problem of the amplification box is not has been solved, and the problem of the mutual recovery of the sealing of the amplification system is solved.

In this embodiment, the number of the amplification chambers 10 is six, the first flow channel 11 and the second flow channel 12 which are connected can form 12 parallel flow channel sections, part of the parallel flow channel sections can be contained in the closed operation area 100, and meanwhile, a plurality of closing auxiliary parts 211 which are arranged at intervals are also contained in the closed operation area 100, and the first flow channel 11 or the second flow channel 12 is arranged between the two closing auxiliary parts 211, so that the first flow channel 11 and the second flow channel 12 which are correspondingly connected in each amplification chamber 10 are separated at intervals, the operation resistance in the closed operation area 100 can be reduced, the deformation of the destructive closure can be contained, the internal stress is released, and the destructive closure is ensured to be more reliable. In this embodiment, six second flow channels 12 are indirectly connected to the system chamber 30 through a common flow channel, the common flow channel comprises a distribution flow channel section 311 at least arranged between two adjacent amplification chambers 10, and of course, the common flow channel comprises a distribution flow channel section 311 which enables even amplification chambers 10 to be equally distributed on two sides, that is, three amplification chambers 10 are respectively arranged on two sides of the distribution flow channel section 311, the common flow channel further comprises a collecting and distributing section which is horizontally arranged, the other end of the distribution flow channel section 311 opposite to the system chamber 30 is connected to the middle part of the collecting and distributing section, the distribution flow channel section 311 and the collecting and distributing section are mutually matched to form a T-shaped structure, the other ends of the six second flow channels 12, which are oppositely connected with the amplification chambers 10, are connected with the flow collecting and distributing section, so that the turbulence degree in the process of transferring the liquid in the chambers 30 to be amplified to the amplification chambers 10 is smaller, and the liquid can be distributed to all the amplification chambers 10 more uniformly and uniformly, and the effect is similar to that of a flute-shaped tube in a fluid conveying scene. The design can enable the flow resistance between different amplification chambers 10 and the system chamber 30 to be basically consistent, ensure that different amplification chambers 10 have basically the same filling state, and enable the subsequent amplification result to be more accurate and reliable. In order to ensure that the amplification chambers 10 can be uniformly and consistently filled, the common flow channel further comprises a shunt buffer section distributed along the width direction of the kit, the other ends of the plurality of second flow channels 12, which are opposite to each other and are connected with the amplification chambers 10, are connected to the shunt buffer section, so that the liquid to be distributed before each amplification chamber 10 is filled can be fully buffered and flows fully developed, the shunt buffer section can be vertically connected with the partial flow section of the distribution flow channel section 311, the hydraulic diameters of the common flow channel, the first flow channel 11 and the second flow channel 12 are in the range of 100 mu m-800 mu m, thus the flow resistance among the chambers of the kit is not excessively large, the processing process of the communication flow channel is ensured to be simple, and the processing precision is higher, the communication flow channel below 100 μm is not selected, the cost is greatly reduced while the processing precision is ensured, the diameter of the common flow channel is configured to be 1.2-3 times that of the first flow channel 11 or the second flow channel 12, more preferably, the hydraulic diameter of the flow channel of the common flow channel is configured to be 1.5-2 times that of the first flow channel 11 or the second flow channel 12, thus ensuring the low turbulence degree distribution of the first flow channel 11 and the second flow channel 12, forming a buffer space for containing liquid to be distributed, ensuring the filling degree of filling in the amplification cavity 10, and the hydraulic diameters of the first flow channel 11 and the second flow channel 12 can be designed to be 300 μm in a specific reagent kit design, whereas the hydraulic diameter of the common flow channel is designed to be 500 μm.

The system cavity 30 of the embodiment has a conical flow guiding portion and a cylindrical accommodating portion, the bottom end of the conical flow guiding portion is provided with a public flow channel interface, the system cavity 30 can have an accommodating space for accommodating a liquid to be dispensed with a preset capacity, the volume of the liquid to be dispensed accommodated in the accommodating space can be 1.2-2 times of the sum of the volumes of all the amplification cavities 10, too much liquid to be dispensed is arranged, the risk of difficult liquid recovery and pollution in the kit is high, and too little liquid is likely to introduce bubbles in the closed amplification cavities 10, so that the reliability of amplification detection results is low. The system cavity 30 is also connected with a buffer cavity 40, the top of the buffer cavity 40 is also provided with an openable connecting cover 42, and can receive samples to be detected or sample treatment fluid to be detected into the buffer cavity 40, here, different types of reagents in different states can be configured in the buffer cavity 40, so that the buffer cavity 40 receives different types of samples, namely, lysate, so that various swab preservation solutions can be used, and blood and tissue solutions are fully mixed with the lysate after waiting for detecting samples to be added into the buffer cavity 40; the present invention is not limited to this, and the buffer chamber 40 and the system chamber 30 are configured to have similar shapes and similar volumes, a buffer communication channel connection port is disposed at the bottom of the buffer chamber 40, the other end of the buffer communication channel is disposed at the bottom of the system chamber 30 and is spaced from a common channel connection port, in order to ensure that the processing operations of the chambers do not affect each other, a fluidic valve 41 is disposed in the buffer communication channel between the buffer chamber 40 and the system chamber 30, a driving gas injection port 32 and a gas vent 43 are disposed at the top of the kit, the driving gas injection port 32 can be communicated with the buffer chamber 40, and the gas vent 43 can be communicated with the system chamber 30.

Fig. 2 is a state diagram of a liquid to be dispensed in a system cavity 30 in a kit, wherein a sample liquid to be detected or a sample processing liquid to be detected in a buffer cavity 40 can be driven by an air source after being completely cracked, a driving air flow is blown in by driving an air injection port 32, and then the liquid can be transferred into the system cavity 30, and can be transferred back and forth between the buffer cavity 40 and the system cavity 30 for several times, so that the liquid in the system cavity 30 can be fully mixed to form the liquid to be dispensed, after the transfer, in combination with the sectional view of fig. 3, a flow control valve 41 is in a closed state, a liquid separation driving part 20 at the bottom is applied with a clamping force by an external clamping device 600, and a deformation recovery piece is compressed and deformed, so that the liquid to be dispensed in the system cavity 30 can be kept in the state of fig. 2, and the operation of applying the clamping force by the external clamping device 600 can also be arranged before the driving the air injection port 32 is blown in the driving air flow, so as to ensure that the amplification cavity 10 is in a positive pressure state and the liquid in the system cavity 30 is not influenced by a fully mixing process.

Fig. 4 and 5 illustrate a process of releasing the liquid separation driving part 20 by the clamping device 600, in which the deformation restoring member in the liquid separation driving part 20 in the clamped and compressed state is gradually restored to the non-compressed state, in which the liquid to be dispensed in the system cavity 30 can be gradually sucked through the common flow channel and the first flow channel 11 to transfer to the amplification cavity 10, the air vent 43 can supplement air to the system cavity 30 to ensure continuous and smooth operation of the transfer process, the liquid separation driving part 20 of the invention comprises liquid separation driving units corresponding to the amplification cavities 10, the filling state of each amplification cavity 10 can be ensured to be substantially the same, and the air and redundant liquid in the amplification cavity 10 can be pumped into the liquid separation driving part 20 through the second flow channel 12, and the whole process of liquid transfer can be gradually and uniformly filled as shown in fig. 6 and 7.

Fig. 8 and 9 are views showing the state where the connection flow path of the filled operation region 100 is closed by the external action, where the filled amplification chamber 10 refers to a state where the filling degree is 95% -100%, the volume relationship between the liquid-dividing driving unit and the amplification chamber 10 needs to be ensured in order to ensure that the state is stable, and at least part or all of the first flow path 11 and the second flow path 12 contain the liquid to be dispensed, and in this state, the external action applied by the closing device 500 may have a destructive action on the closed operation region 100 while the first flow path 11 and the second flow path 12 are destructively closed, and where the closing device 500 of the external action may be at least one of the forms of external action by gluing, extrusion deformation, thermal fusion, welding, fluid curing, etc., and destructively closing the first flow path 11 and the second flow path 12. The closure device 500 may be a pressurizing device that breaks the communication relationship of the flow channels by squeezing deformation; the communication relation of the flow channels can be destroyed by filling and extruding the foaming agent through the foaming device; can be a thermal welding device, and the communicating relation of the runners is destroyed by fusing the runner constituent materials at high temperature; the device can be laser welding equipment, and the communication relation of the flow channels is destroyed by high-temperature fusion welding of laser; the device can be light wave emitting equipment such as ultraviolet, and the communication relation of the flow channels is destroyed by solidifying specific fluid by ultraviolet and the like. Fig. 10 is a schematic diagram of the state of the first flow channel 11 and the second flow channel 12 of the amplification chamber 10 before filling, fig. 11 is a schematic diagram of the state of the first flow channel 11 and the second flow channel 12 of the amplification chamber 10 after filling, the previous explanation has been made about the liquid filling process of the amplification chamber 10, the closing device 500 can act on the closed operation area 100 after filling, further, a destructive closing effect is applied to the first flow channel 11 and the second flow channel 12 at the same time, so as to realize the simultaneous efficient closing of at least two filling state amplification chambers 10, the first flow channel 11 and the second flow channel 12 are arranged in parallel flow channel sections in the closed operation area 100, and the closed strengthening part 210 is arranged at the position where the closed strengthening part 210 can reduce the flow cross section to 1/3-2/3 of the flow channel cross section at the position where the closed strengthening part is not arranged, so that the flow cross section is required to be broken, the closed operation area is smaller, the energy consumption or material consumption is reduced, and the first flow channel 11 and the second flow channel 12 can be closed more reliably, and the problem that the flow cross section is excessively large or the flow dead zone is caused by the arrangement.

The closing reinforcement portion 210 and the closing auxiliary portion 211 may cooperate with each other in the closing operation region 100 to form a linear operation region having a predetermined width, the linear operation region is perpendicular to the parallel flow path section, the communication flow path in the linear operation region may be completely destructively closed under the external action, where the predetermined width of the linear operation region may be between 0.3mm and 2mm, and the linear operation region is not too large, otherwise, the state and stability of filling the amplification chamber 10 with the liquid may be affected, and the problem that the communication flow path cannot be reliably closed is not too small.

Example 2

An amplification apparatus comprising the kit provided in example 1, further comprising

A kit receiving location capable of receiving and confining the kit;

A clamping device 600 which can apply clamping or releasing operation to the liquid separation driving part 20 to enable the liquid separation driving part 20 to generate pressure change and drive the liquid to be dispensed in the system cavity 30 to be filled in the amplification cavity 10 through the second flow channel 12;

a closure device 500 capable of applying a destructive closure operation to the first flow channel 11 and the second flow channel 12;

a heating module capable of performing a cyclic heating operation on the closed amplification chamber 10;

The detection module can cooperate with the heating module to perform fluorescence acquisition operation on the sealed amplification cavity 10 in the cyclic heating process, so as to obtain an amplification detection result.

Example 3

A closed amplification detection method is applied to the amplification equipment provided in the embodiment 2, and comprises the following steps,

Filling: starting the liquid separation driving part 20 to generate pressure difference between the amplification cavity 10 and the system cavity 30, and driving the liquid to be distributed in the system cavity 30 to be filled in the amplification cavity 10 to obtain the filled amplification cavity 10;

And (3) a sealing step: under the external action of the sealing device 500, the first flow channel 11 and the second flow channel 12 are simultaneously and destructively sealed, so that the filling-state amplification cavity 10 is isolated and sealed, and the liquid in the filling-state amplification cavity 10 is used as an amplification system liquid;

amplification detection step: and amplifying the amplification system liquid in the amplification cavity 10 in a closed state, and performing fluorescent detection on the amplification system liquid in the amplification to obtain an amplification detection result.

The principles and embodiments of the present invention have been described herein with reference to specific examples, which are intended to be merely illustrative of the methods of the present invention and their core ideas. It should be noted that it will be apparent to those skilled in the art that various modifications and adaptations of the invention can be made without departing from the principles of the invention and these modifications and adaptations are intended to be within the scope of the invention as defined in the following claims.

In the description of the present invention, it should be noted that, directions or positional relationships indicated by terms such as "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc., are directions or positional relationships based on those shown in the drawings, or are directions or positional relationships conventionally put in use of the inventive product, are merely for convenience of describing the present invention and simplifying the description, and are not indicative or implying that the apparatus or element to be referred to must have a specific direction, be constructed and operated in a specific direction, and thus should not be construed as limiting the present invention.

In the description of the present invention, it should also be noted that, unless explicitly specified and limited otherwise, the terms "disposed," "mounted," "connected," and "connected" are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communication between two elements. The specific meaning of the above terms in the present invention will be understood in specific cases by those of ordinary skill in the art.

Claims (10)

1. The utility model provides a kit, its characterized in that, includes the kit body, this internal system chamber, amplification chamber, divide liquid drive portion and the intercommunication runner of being equipped with of kit, the intercommunication runner includes first runner and second runner, the one end of first runner communicates in the top of amplification chamber, the other end communicates divides liquid drive portion, the one end of second runner communicates in the middle part of amplification chamber, the other end intercommunication system chamber, divide liquid drive portion accessible pressure variation drive system intracavity wait to distribute liquid to fill in amplification chamber through the second runner, first runner and second runner can be simultaneously by destructive closure.

2. The kit according to claim 1, wherein the kit body is further provided therein with a closing operation area for destructively closing the first flow channel and the second flow channel, a plurality of closing aids are arranged in the closing operation area, and the plurality of closing aids are arranged between the first flow channel and the second flow channel at intervals so that the first flow channel and the second flow channel are separated at intervals.

3. The kit according to claim 2, wherein the communication flow path includes a parallel flow path section matching the length direction of the kit, the parallel flow path section including a first flow path parallel section corresponding to the first flow path, and a second flow path parallel section corresponding to the second flow path, at least part of the first flow path parallel section and the second flow path parallel section being arranged in the closed operation region.

4. The kit according to claim 3, wherein the parallel flow channel section is provided with a closing reinforcement portion at a position corresponding to the closing operation region, and a flow channel sectional area of the parallel flow channel section where the closing reinforcement portion is provided is smaller than a flow channel sectional area of the parallel flow channel section where the closing reinforcement portion is not provided.

5. The kit according to claim 1, wherein the liquid separation driving part is provided with a deformation recovery piece, the deformation recovery piece can deform when being extruded by external force, and can recover and generate suction force when no external force acts, so that the liquid to be distributed in the system cavity is filled in the amplification cavity through the second flow channel.

6. The kit according to claim 1, wherein the region above the centerline of the amplification chamber is a contoured portion having a gradually decreasing cross-sectional area from bottom to top.

7. The kit according to claim 1, wherein a buffer cavity is further arranged in the kit body, the buffer cavity is communicated with the system cavity through a buffer communication flow channel, and a flow control valve is arranged at the buffer communication flow channel; the buffer cavity is also provided with an openable connecting cover, the buffer cavity receives external liquid into the buffer cavity through the connecting cover, the kit body is also provided with a driving gas injection port and a vent port, the driving gas injection port is communicated with the buffer cavity and can blow driving gas flow into the buffer cavity, and the vent port is communicated with the system cavity and can supplement gas into the system cavity.

8. The kit according to any one of claims 1 to 7, wherein the number of the amplification chambers is an even number, liquid-dividing driving units matched with the number of the amplification chambers are arranged in the liquid-dividing driving part, each amplification chamber is communicated with the corresponding liquid-dividing driving unit through a corresponding first flow channel and is communicated with a common flow channel of the system chamber through a corresponding second flow channel, and the even number of amplification chambers are symmetrically distributed on two sides of a distribution flow channel section of the common flow channel.

9. An amplification apparatus comprising the kit of any one of claims 1-8, further comprising

A kit receiving location capable of receiving and confining the kit;

the clamping device can apply clamping or releasing operation to the liquid separation driving part to enable the liquid separation driving part to generate pressure change and drive liquid to be distributed in the system cavity to be filled in the amplification cavity through the second flow channel;

A closure device capable of applying a destructive closure operation to the first flow path and the second flow path;

a heating module capable of performing a cyclic heating operation on the closed amplification chamber;

The detection module can cooperate with the heating module to perform fluorescence acquisition operation on the sealed amplification cavity in the cyclic heating process, so as to obtain an amplification detection result.

10. A method for detecting the amplification of a closed state, which is applied to the amplification equipment according to claim 9, comprises the following steps,

Filling: starting a liquid separation driving part to generate pressure difference between the amplification cavity and the system cavity, and driving liquid to be distributed in the system cavity to be filled in the amplification cavity to obtain a filled amplification cavity;

and (3) a sealing step: under the external action applied by the sealing device, the first flow channel and the second flow channel are simultaneously and destructively sealed, so that the filling-state amplification cavity is isolated and sealed, and the liquid in the filling-state amplification cavity is used as the amplification system liquid;

amplification detection step: amplifying the amplification system liquid in the amplification cavity in a closed state, and carrying out fluorescent detection on the amplification system liquid in the amplification to obtain an amplification detection result.