CN218842179U - Nucleic acid amplification device and nucleic acid detection device - Google Patents

Abstract

The invention provides a nucleic acid amplification device, which comprises a rotary disc, a rotary disc driving die, a temperature control module, an infrared heater, a lateral telescopic rod mechanism and a shell, wherein the shell surrounds the rotary disc driving module; the temperature control module comprises a bottom plate and a heating column arranged on the bottom plate, a through hole is formed in the bottom plate, the temperature control module is sleeved on the shell through the through hole, and the infrared heater is arranged at the top of the shell; the lateral telescopic rod mechanism is fixed on the side wall of the shell, and the top end of a telescopic rod of the lateral telescopic rod mechanism is connected with the bottom plate. The device disclosed is short in detection time and high in efficiency.

Description

Nucleic acid amplification device and nucleic acid detection device

Technical Field

The present disclosure relates to the field of in vitro detection, and in particular, to a nucleic acid amplification device and a nucleic acid detection device.

Background

A variable-temperature amplification method relates to the earliest nucleic acid amplification technology, and PCR is characterized in that DNA is denatured into single strands at a high temperature of 95 ℃ in vitro, primers and the single strands are combined according to the base complementary pairing principle at a low temperature (usually about 60 ℃), the temperature is adjusted to the optimal reaction temperature (about 72 ℃) of DNA polymerase, and the DNA polymerase synthesizes a complementary strand along the direction from phosphoric acid to pentose (5 '-3'). The PCR instrument manufactured based on polymerase is actually a temperature control device, and can be well controlled among denaturation temperature, renaturation temperature and extension temperature.

However, the conventional nucleic acid detection method is long in time and high in cost, and thus, the daily nucleic acid detection becomes a burden. Therefore, it has been difficult in the industry to find a method and a system for nucleic acid amplification detection that can be operated quickly, inexpensively and automatically by combining the rapid nucleic acid amplification detection with the rapid nucleic acid detection equipment of fluorescence detection technology.

SUMMERY OF THE UTILITY MODEL

In order to solve the problems in the related art, the present disclosure provides a nucleic acid amplification apparatus and a nucleic acid detection apparatus.

According to a first aspect of the present disclosure, a nucleic acid amplification device in the present disclosure includes a turntable, a turntable driving die, a temperature control module, an infrared heater, a lateral telescopic rod mechanism, and a housing, the housing encloses the turntable driving module, a through hole is provided at the top of the housing, and a rotating shaft of the turntable driving module extends out from the top of the housing through the through hole and is connected to the turntable through a center hole of the turntable; the temperature control module comprises a bottom plate and a heating column arranged on the bottom plate, a through hole is formed in the bottom plate, the temperature control module is sleeved on the shell through the through hole, and the infrared heater is arranged at the top of the shell; the lateral telescopic rod mechanism is fixed on the side wall of the shell, and the top end of a telescopic rod of the lateral telescopic rod mechanism is connected with the bottom plate.

Illustratively, the heating column is composed of a heat conducting aluminum block, an electromagnetic induction heating coil and a heat insulating column, wherein the heat insulating column is arranged on the bottom plate, the heat conducting aluminum block is connected with the top end of the heat insulating column, and the electromagnetic induction heating coil surrounds the heat conducting aluminum block.

As an example, the number of the heating columns is 3N, where N is a natural number.

As an example, the number of the heating columns is 9.

As an example, the top of the heat conductive aluminum block has a groove.

Illustratively, the groove has a circular cavity and a tapered cavity.

Illustratively, the lateral telescopic rod mechanism further comprises a housing and a lifting motor, wherein the lifting motor is connected with the telescopic rod to drive the telescopic rod to move up and down, and the telescopic rod extends out of the top of the housing and is connected with the bottom plate of the temperature control module.

Illustratively, the turntable driving module includes a motor, a fixed pin bearing, and a rotation shaft.

According to a second aspect of the present disclosure, there is provided a nucleic acid detection device, comprising the aforementioned nucleic acid amplification device, and a fluorescence collection module

Illustratively, the fluorescence collection module comprises an excitation light source and a photodetector, and the excitation light source and the photodetector are vertical above the turntable.

Advantageous technical effects

The device that this disclosure provided adopts the aluminium pig heating that the heat conductivility is high, and 9 heat conduction aluminium pigs are through being controlled by temperature control module, according to clockwise stable at "denaturation, annealing and extension" required temperature, can satisfy PCR nucleic acid amplification's temperature cycle. While the traditional PCR tube needs to heat and cool the reaction tube repeatedly for nucleic acid amplification, which greatly consumes time. In addition, the light wave emitted by the fluorescence detection module excitation light source is reflected to the photoelectric detector through the V-shaped aluminum block groove, so that the fluorescence can be collected; meanwhile, the turntable can simultaneously amplify the nucleic acid of three microfluidic chips in real time on line by one-time operation, and if the size of the turntable is not considered, the chips can be continuously added onto the turntable, so that the detection efficiency is further improved.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the disclosure.

Drawings

Other features, objects, and advantages of the present disclosure will become more apparent from the following detailed description of non-limiting embodiments when taken in conjunction with the accompanying drawings. The following is a description of the drawings.

FIG. 1 shows a block diagram of a nucleic acid amplification apparatus according to the present disclosure.

Fig. 2 shows a structure of a microfluidic chip according to the present disclosure.

Fig. 3 shows a structural view of a plastic encapsulation film of a microfluidic chip according to the present disclosure.

Fig. 4 shows a block diagram of a chip body of a microfluidic chip according to the present disclosure.

Fig. 5 shows a side view of a chip body of a microfluidic chip according to the present disclosure.

Fig. 6 shows a block diagram of a turntable according to the present disclosure.

FIG. 7 illustrates a block diagram of a temperature control module according to the present disclosure.

Fig. 8 shows a temperature arrangement schematic of a temperature control module according to the present disclosure.

Fig. 9 illustrates a structural view of a groove of a thermally conductive aluminum block according to the present disclosure.

Fig. 10 shows a block diagram of a turntable drive module according to the present disclosure.

Figure 11 illustrates a side extension pole mechanism configuration according to the present disclosure.

It should be understood that the dimensions of the various elements shown in the figures are not drawn to scale. Further, the same or similar reference numerals denote the same or similar components.

Detailed Description

Hereinafter, exemplary embodiments of the present disclosure will be described in detail with reference to the accompanying drawings so that those skilled in the art can easily implement them. Also, for the sake of clarity, parts not relevant to the description of the exemplary embodiments are omitted in the drawings.

In the present disclosure, it is to be understood that terms such as "including" or "having," etc., are intended to indicate the presence of the disclosed features, numbers, steps, behaviors, components, parts, or combinations thereof, and are not intended to preclude the possibility that one or more other features, numbers, steps, behaviors, components, parts, or combinations thereof may be present or added.

The same or similar reference numerals in the drawings of the present disclosure correspond to the same or similar components; in the description of the present disclosure, it is to be understood that, if there are terms such as "center", "upper", "lower", "left", "right", "horizontal", "inner", "outer", etc., indicating orientations or positional relationships based on the orientations or positional relationships shown in the drawings, the description is merely for convenience in describing the present disclosure and simplifying the description, but not for indicating or implying that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation, and thus the positional relationships described in the drawings are for illustrative purposes only and are not to be construed as limitations of the present disclosure, and specific meanings of the terms may be understood by those skilled in the art according to specific circumstances. Furthermore, the terms "first," "second," "third," and the like are used merely to distinguish one element from another, and are not to be construed as indicating or implying relative importance.

Throughout the description of the present disclosure, it should also be noted that, unless expressly stated or limited otherwise, the terms "disposed," "mounted," "connected," and "coupled" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; either directly or indirectly through intervening media, or may be interconnected between two elements. The specific meaning of the above terms in the present disclosure can be understood in a specific case to those of ordinary skill in the art.

It should be further noted that the embodiments and features of the embodiments in the present disclosure may be combined with each other without conflict. The present disclosure will be described in detail below with reference to the accompanying drawings in conjunction with embodiments.

As mentioned above, the current nucleic acid detection is long and costly, and the daily nucleic acid detection is a burden, the present disclosure provides a nucleic acid amplification apparatus in order to solve the problem.

As shown in fig. 1, the nucleic acid amplification device in the present disclosure includes a turntable 1, a turntable driving module, a temperature control module 3, a housing 4, a lateral telescopic rod mechanism 5, and an infrared heater 6, wherein the housing 4 surrounds the turntable driving module, a through hole 41 is formed in the top of the housing 4, and a rotating shaft 71 of the turntable driving module extends out of the top of the housing 4 through the through hole and is connected to the turntable 1 through a central hole 104 of the turntable 1; the temperature control module 3 comprises a bottom plate 305 and a heating column arranged on the bottom plate 305, a through hole 304 is arranged on the bottom plate 305, the temperature control module 3 is sleeved on the shell 4 through the through hole 304, and the infrared heater 6 is arranged on the top of the shell 4; the lateral telescopic rod mechanism is fixed on the side wall of the housing 4, and the top end of the telescopic rod of the lateral telescopic rod mechanism is connected with the bottom plate 305.

In the present disclosure, the housing 4 encloses the turntable driving module to protect the motor of the turntable driving module, the rotating shaft 71 of the turntable driving module can drive the turntable 1 to rotate, the turntable 1 is loaded with a microfluidic chip for nucleic acid amplification reaction, and the rotation of the turntable 1 can drive the liquid in each chamber of the microfluidic chip to flow so as to complete the amplification reaction (which will be discussed in detail later). The temperature control module 3 can heat the microfluidic chip 2 to meet the temperature requirement of the amplification reaction. The infrared heater is aligned with the wax valve chamber of the microfluidic chip and used for heating the wax valve chamber. The lateral telescopic mechanism moves up and down through the telescopic rod to drive the temperature control module 3 to move up and down.

As mentioned above, the rotation of the rotating disc 1 can drive the liquid in each chamber of the microfluidic chip 2 to flow so as to complete the amplification reaction. Specifically, as shown in fig. 2 to 5, the microfluidic chip 2 includes a plastic package film 10 and a chip main body 20, the chip main body 20 is provided with a sample adding cavity 201, a sample pre-storage cavity 203, a wax valve cavity 210, a first vent 206, a second vent 207, and a reaction chamber 212 protruding downward, wherein the sample adding cavity 201 is communicated with the sample pre-storage cavity 203 through a first flow channel 202, the sample pre-storage cavity 203 is communicated with the reaction chamber 212 through a second flow channel 205, the wax valve cavity 210 is communicated with the reaction chamber 212 through a third flow channel 211, the first vent 206 is communicated with the reaction chamber 212 through a fourth flow channel 214, and the second vent 207 is communicated with the wax valve cavity 210 through a fifth flow channel 208. The sample adding cavity 201, the sample pre-storage cavity 203, the wax valve cavity 210, the first vent hole 206, the second vent hole 207, the reaction cavity 212 protruding downwards, the first flow channel 202, the second flow channel 205, the third flow channel 211 and the fourth flow channel 214 are processed on the upper surface of the chip main body 20 through the prior art such as micro-nano processing and the like, and then are sealed on the upper surface of the chip main body 20 through the plastic packaging film 10 so as to provide closed cavities and flow channels. The sample application chamber 201 is used for sample application and also as a vent for gas venting during liquid flow. The sample pre-storage cavity 203 is used for storing a sample, and after the sample is added from the sample adding cavity 201, the sample directly flows into the sample storage cavity 203 through the first flow channel 202 to be subjected to subsequent reaction. The wax valve cavity 210 is used for pre-storing paraffin oil 209 in a solid state, the melted paraffin oil 209 enters the reaction cavity 212 from the wax valve cavity 210 through the third flow passage 211 under the action of centrifugal force, and the paraffin oil 209 is used for preventing the reaction liquid from evaporating in the reaction process. The reaction chamber 212 is used for PCR reaction. The reaction chamber 212 is provided with pre-stored PCR reagent freeze-dried beads 213 for completing the PCR amplification reaction after the sample and paraffin oil enter the reaction chamber 212. The reaction chamber 212 includes a circular member 2121 protruding downward from the lower surface of the chip body 2, and a tapered member 2122, and the tapered member 2122 is coupled to the circular member 2121 to form the reaction chamber 212. By disposing the reaction chamber 212 to protrude downward from the lower surface of the chip body 2, the thermal cycling conditions for PCR amplification are more uniform and efficient in heat transfer than the flat plate type chip. Illustratively, cone 2122 has a "V" shaped configuration. Be provided with three exhaust hole on the plastic envelope membrane 1, be the first exhaust hole 101 of plastic envelope membrane, plastic envelope membrane second exhaust hole 102 and plastic envelope membrane third exhaust hole 103 respectively, three exhaust hole respectively with application of sample chamber 201, first exhaust hole 206, second exhaust hole 207 correspond the setting. When the device is used, due to the fact that liquid flows, gas needs to be discharged, three exhaust holes are formed in the plastic packaging film 1, and the three exhaust holes correspond to the sample adding cavity 201, the first exhaust hole 206 and the second exhaust hole 207 respectively and can discharge gas in the chip so as to facilitate the flow of the liquid.

Referring to fig. 6, the turntable 1 is provided with a circular through hole 104 in the middle and at least one recess 101, the shape of the recess 101 being adapted to the microfluidic chip to secure the microfluidic chip in the recess 101. As an example, as shown in fig. 6, the number of grooves may be 3. As mentioned above, the rotating shaft 71 of the turntable driving module can drive the turntable 1 to rotate, and centrifugal force is generated during the rotation of the turntable 1, and the centrifugal force causes the liquid in the chamber of the chip body 2 to flow into the reaction chamber for PCR amplification reaction.

As shown in fig. 10, the turntable driving module 7 includes a motor 74, a fixing pin 73, a bearing 72, and a rotating shaft 71, wherein the fixing pin 73 connects the bearing 72 with the rotating shaft 71 to connect with the motor, so that the motor can drive the rotating shaft 71 to rotate. As an example, the motor may be a servo motor. The turntable driving module 7 is driven by a motor controller, and the motor controller is provided with a rotary encoder and can control the motor to rotate. The motor controller may be a commercially available product and will not be described herein.

As mentioned above, the infrared heater is aligned with the wax valve chamber of the microfluidic chip for heating the wax valve chamber, and specifically, the turntable 1 is provided with the infrared irradiation hole 103, and the infrared heater, the wax valve chamber and the infrared irradiation hole 103 are aligned, so that the wax valve chamber of the microfluidic chip is aligned for heating.

Illustratively, as shown in fig. 7, the heating column is composed of a heat conducting aluminum block 301, an electromagnetic induction heating coil 302, and a heat insulating column 303, wherein the heat insulating column 303 is disposed on a bottom plate 305, the heat conducting aluminum block 301 is connected to a top end of the heat insulating column 303, and the electromagnetic induction heating coil 302 surrounds the heat conducting aluminum block 301 to heat the heat conducting aluminum block. As an example, the temperature control module 3 adopts an eddy current heating mode, the control method is PID control, the lower coil heats the heat conducting aluminum block under the action of alternating current, and the heat conducting aluminum block is provided with a temperature sensor for monitoring the temperature of the heat conducting aluminum block in real time and keeping the temperature of the heat conducting aluminum block to reach the required PCR temperature.

As an example, the number of the heating columns is 3N, where N is a natural number. In a specific embodiment, as shown in FIGS. 7 to 8, the number of the heating columns is 9, every adjacent three are used as a group, and the heating temperatures are 95 ℃, 72 ℃ and 65 ℃ respectively as the denaturation temperature, the extension temperature and the annealing temperature in the PCR process, as shown in FIG. 8. Of course, one skilled in the art will also appreciate that other temperatures may be used so long as denaturation, extension, and annealing are achieved.

Illustratively, as shown in fig. 9, the top of the heat conducting aluminum block 301 has a groove 3011 to accommodate the reaction of the microfluidic chip, so as to heat the reaction chamber 212. Reaction wells 102 are also provided in the grooves 101 to extend the reaction chambers 212 of the microfluidic chip from the bottom of the carousel 1 to be received in the grooves 3011 of the heating posts.

Illustratively, the recess 3011 has a circular cavity 30111 and a tapered cavity 30112 to match the shape of the reaction chamber 212, so that the recess 3011 and the reaction chamber 212 can be more closely fitted to improve temperature conduction. By way of example, the tapered cavity 30112 may be V-shaped.

Illustratively, as shown in fig. 11, the lateral expansion link mechanism 5 further includes a lifting motor 51, an electric push rod main body 52, and a push rod 53, wherein the lifting motor 51 can drive the electric push rod main body 52 to move up and down with the expansion link 53, and the expansion link 53 is connected to a bottom plate 305 of the temperature control module 3. The lateral telescopic mechanism 5 is fixed on the shell 4 to save space and reduce the volume of the equipment. The telescopic link can drive temperature control module 3 and reciprocate, and is concrete, during the rebound, thereby reaction chamber 212 is held in the recess 3011 of heating post and is heated it, and when the rebound, reaction chamber 212 separates rather than in the recess 3011 of heating post, and the motor can drive pivot 71 and rotate and then make carousel 1 rotate to make reaction chamber 212 get into the position of next heating post.

According to a second aspect of the present disclosure, the present disclosure also provides a nucleic acid detecting device to achieve rapid detection. According to the present disclosure, a nucleic acid detection device is provided, which includes a nucleic acid amplification device provided in the first aspect of the present disclosure, and a fluorescence collection module.

Illustratively, the fluorescence collection module includes an excitation light source and a photodetector, which are vertical above the turntable 1. In the present disclosure, the fluorescence collecting module may be any commercially available fluorescence collecting module as long as the fluorescence intensity can be detected, which is not described herein again.

According to the nucleic acid detecting device provided by the present disclosure, the use steps are as follows:

1. adding a nucleic acid solution to be detected into a microfluidic chip, wherein a pre-stored freeze-dried reagent is arranged in a chip sample pre-storage chamber, and pre-stored wax is arranged in a wax valve chamber;

2. the motor controller controls the motor to rotate at a low speed, the rotary encoder works, an initialization positioning signal is sent to the rotary encoder, the motor stops, and initialization positioning is completed;

3. heating the wax storage cavity hole on the turntable by using an infrared heater, wherein the solid wax is changed into liquid wax after 5 seconds;

4. the motor controller controls the motor to rotate at a high speed, so that the nucleic acid solution and the melted wax solution are automatically mixed in the reaction chamber in a high-speed centrifugal manner, and a freeze-drying reagent is pre-embedded in the reaction chamber, specifically, the rotating speed is 2000-3000 r/min;

5. simultaneously, as shown in fig. 8, the temperature control module adopts a variable-width pulse signal to enable the 9 electromagnetic induction heating coils to respectively heat and arrange the 9 heat-conducting aluminum blocks in a manner of 95 ℃, 60 ℃, 72 ℃, 95 ℃, 60 ℃ and 72 ℃ (clockwise);

6. the motor stops rotating, the turntable returns to the initial position, the lateral telescopic rod mechanism drives the heat conduction aluminum block to move upwards for 3cm, so that the reaction chambers of the 3 micro-fluidic chips just fall into the 3V-shaped grooves of the heat conduction aluminum block with the temperature of 95 ℃, and the reaction chambers are subjected to denaturation reaction for 15 seconds at the temperature of 95 ℃. After finishing, the telescopic rod mechanism is laterally slid to enable the turntable to return to the initial position by 3cm downwards

7. The motor is controlled by a rotary encoder to drive the rotary table to rotate 120 degrees clockwise, 3 chips on the rotary table are alternately dropped on 3 constant-temperature heat-conducting aluminum blocks at 60 ℃, the lateral telescopic rod mechanism drives the heat-conducting aluminum blocks to move upwards for 3cm again, reaction chambers of the 3 micro-fluidic chips are just dropped in the V-shaped grooves of the 3 heat-conducting aluminum blocks at 60 ℃, and after the reaction chambers are annealed for 30 seconds, the lateral sliding telescopic rod mechanism enables the rotary table to return 3cm downwards to the initial position;

8. the motor is controlled by a rotary encoder to drive the turntable to rotate 120 degrees clockwise, reaction chambers of 3 micro-fluidic chips on the turntable are alternately made to fall onto 3 72-DEG C constant-temperature heat-conducting aluminum blocks, the lateral telescopic rod mechanism drives the heat-conducting aluminum blocks to move upwards for 3cm again, the reaction chambers of the 3 micro-fluidic chips are made to just fall into 3V-shaped grooves of the 72-DEG C heat-conducting aluminum blocks, and after the reaction chambers are extended and react for 45 seconds, the lateral sliding telescopic rod mechanism enables the turntable to return 3cm downwards to the initial position;

9. repeating the steps (6), (7) and (8) for 30-40 times to complete the whole nucleic acid amplification link;

10. the motor controller controls the motor to drive the turntable to reset, the excitation light source emits light waves, excited fluorescence is reflected to the photoelectric detector, the photoelectric detector generates analog signals, and the analog signals are sent to the signal acquisition processor to generate real-time fluorescence detection signals;

11. and controlling a motor to drive the next reaction chamber on the turntable to an excitation light source irradiation hole through a motor controller to complete the fluorescence signal detection of the second micro-fluidic chip and sequentially complete the fluorescence detection of the third micro-fluidic chip.

The foregoing description is only exemplary of the preferred embodiments of the disclosure and is illustrative of the principles of the technology employed. It will be understood by those skilled in the art that the scope of the present invention is not limited to the specific combination of the above-mentioned features, but also covers other embodiments formed by any combination of the above-mentioned features or their equivalents without departing from the spirit of the present invention. For example, the above features and the technical features disclosed in the present disclosure (but not limited to) having similar functions are replaced with each other to form the technical solution.

Claims (10)

1. A nucleic acid amplification device is characterized by comprising a rotary disc, a rotary disc driving die, a temperature control module, an infrared heater, a lateral telescopic rod mechanism and a shell, wherein the shell surrounds the rotary disc driving module, a through hole is formed in the top of the shell, and a rotating shaft of the rotary disc driving module extends out of the top of the shell through the through hole and is connected with the rotary disc through a central hole of the rotary disc; the temperature control module comprises a bottom plate and a heating column arranged on the bottom plate, a through hole is formed in the bottom plate, the temperature control module is sleeved on the shell through the through hole, and the infrared heater is arranged at the top of the shell; the lateral telescopic rod mechanism is fixed on the side wall of the shell, and the top end of a telescopic rod of the lateral telescopic rod mechanism is connected with the bottom plate.

2. The nucleic acid amplification apparatus of claim 1, wherein the heating column comprises a heat conductive aluminum block, an electromagnetic induction heating coil, and a heat insulating column, wherein the heat insulating column is disposed on the bottom plate, the heat conductive aluminum block is connected to a top end of the heat insulating column, and the electromagnetic induction heating coil surrounds the heat conductive aluminum block.

3. The nucleic acid amplification apparatus according to claim 2, wherein the number of the heating columns is 3N, where N is a natural number.

4. The nucleic acid amplification apparatus according to claim 3, wherein the number of the heating columns is 9.

5. The nucleic acid amplification apparatus according to any one of claims 2 to 4, wherein the heat conductive aluminum block has a groove at the top.

6. The nucleic acid amplification apparatus of claim 5, wherein the groove has a circular cavity and a tapered cavity.

7. The nucleic acid amplification apparatus of claim 1, wherein the lateral expansion link mechanism further has a housing and a lifting motor connected to the expansion link to drive the expansion link to move up and down, the expansion link extending from a top of the housing and connected to a bottom plate of the temperature control module.

8. The nucleic acid amplification apparatus of claim 1, wherein the turntable driving module comprises a motor, a fixed pin bearing, and a rotating shaft.

9. A nucleic acid detecting device comprising the nucleic acid amplification device of any one of claims 1-8, and a fluorescence collection module.

10. The nucleic acid detecting device according to claim 9, wherein the fluorescence collecting module comprises an excitation light source and a photodetector, and the excitation light source and the photodetector are vertical above the turntable.

Images