CN113736638A - PCR (polymerase chain reaction) heating system, device and method - Google Patents

Abstract

The invention provides a PCR (polymerase chain reaction) warming and heating system, a device and a method, wherein the system comprises a TEC heating module, a radiator, a cooling fan and a PCR amplification reaction container; the radiator is fixedly connected to a first surface of the TEC heating module, the radiator fan is arranged on a second surface adjacent to the first surface, and the PCR amplification reaction container is in movable contact with the radiator; on one hand, the system has simple integral structure and wide application range, can be installed and used in an integrated manner, and can realize high-precision rapid temperature rise and drop; on the other hand, compared with the existing PCR heating scheme, the PCR heating system in the technical scheme of the application can effectively control the production cost due to the simple structure, has stronger practicability and expansibility, and can be widely applied to the technical field of biology.

Description

PCR (polymerase chain reaction) heating system, device and method

Technical Field

The invention relates to the technical field of molecular biology, in particular to a PCR heating system, a device and a method.

Background

PCR (Polymerase Chain Reaction) is the key to the success or failure of detection when amplifying and detecting nucleic acid, and consists of three steps of denaturation, annealing and extension, and the specificity of the PCR depends on oligonucleotide primers which are complementary with both ends of a target sequence and is similar to the natural replication process of DNA. Different reaction stages of PCR require different holding temperatures, DNA is denatured into single strands at 95 ℃, primers and single strands are combined according to the base complementary pairing principle at about 60 ℃, the most suitable reaction temperature of DNA polymerase is reached when the temperature is adjusted to about 72 ℃, and the DNA polymerase catalyzes and synthesizes complementary strands from a hydroxyl end to a phosphate group along a template according to the base complementary pairing principle. Therefore, the temperature needs to be controlled rapidly and precisely during the PCR process.

Because the existing PCR heating device mostly puts the reaction solvent into an oil bath and a metal bath, and the oil bath and the metal bath are mostly used for indirectly supplying temperature to the reaction solvent, the reaction solvent has delay property; the existing PCR heating device has the problems of heat loss and inaccuracy of temperature control in the heat transfer process. In addition, the conventional PCR heating apparatus has poor accuracy, complicated structure and low practicability.

Disclosure of Invention

In view of the above, to at least partially solve one of the above technical problems, embodiments of the present invention provide a PCR temperature-increasing heating system with higher accuracy and faster temperature control speed, and a corresponding apparatus and control method.

In a first aspect, the present application provides a PCR warming and heating system, which includes: the TEC heating module, the radiator, the cooling fan and the PCR amplification reaction container;

the radiator is fixedly connected to a first surface of the TEC heating module, the radiator fan is arranged on a second surface adjacent to the first surface, and the PCR amplification reaction container is in movable contact with the radiator;

the TEC heating module is used for generating heat; the heat radiator is used for conducting the heat to the PCR amplification reaction vessel; the PCR amplification reaction container is used for heating the PCR amplification reagent by the heat obtained by conduction of the radiator; the heat dissipation fan is used for guiding the heat dissipation airflow of the TEC heating module.

In one possible embodiment of the present disclosure, the system further comprises a temperature sensor disposed on a surface of the PCR amplification reaction vessel.

In one possible embodiment of the present disclosure, the heat dissipation fan includes a first heat dissipation fan and a second heat dissipation fan; the first heat dissipation fan is arranged on a second surface adjacent to the first surface, and the second heat dissipation fan is arranged on a third surface far away from the second surface.

In a feasible embodiment of the scheme of the application, the surface of the heat radiator away from the PCR amplification reaction container is provided with at least one groove, the grooves are parallel to each other, and the grooves are used for placing a centrifugal tube or a chip-type flow channel.

In a possible embodiment of the present disclosure, the TEC heating module includes a hot end and a cold end; a first metal conductor is arranged on the surface of the hot end close to the cold end, a second metal conductor is arranged on the surface of the cold end close to the hot end, and a plurality of semiconductor elements are arranged between the first metal conductor and the second metal conductor; the hot end and the cold end are ceramic electrodes.

In a possible embodiment of the present disclosure, the groove includes a communication groove for storing the PCR amplification reagents after mixing; the TEC heating module is also connected with an external air pump, and the external air pump is used for controlling the PCR amplification rate.

In one possible embodiment of the protocol of the present application, the PCR amplification reaction vessel is filled with a solution for storing heat and/or transferring heat.

In a second aspect, the present invention further provides a PCR warming-up heating apparatus, which includes any one of the PCR warming-up heating systems described in the first aspect:

in a third aspect, the present invention further provides a PCR warming-up heating method for controlling any one of the PCR warming-up heating systems described in the first aspect, including the following steps:

controlling the TEC heating module to heat to generate heat;

and conducting the heat to the PCR amplification reaction container through the heat radiator, so that the PCR amplification reaction container conducts the obtained heat to heat the PCR amplification reagent through the heat radiator.

In a possible embodiment of the solution of the present application, the method further comprises the steps of:

and guiding the heat dissipation airflow of the TEC heating module by a heat dissipation fan.

Advantages and benefits of the present invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention:

according to the technical scheme, the system mainly comprises a TEC heating module, a radiator, a cooling fan, a PCR amplification reaction container and other components, on one hand, the system is simple in overall structure, wide in application range, capable of being installed and used in an integrated mode, and capable of achieving high-precision rapid temperature rise and drop; on the other hand, compared with the existing PCR heating scheme, the PCR heating system in the technical scheme of the application can effectively control the production cost due to the simple structure, and has stronger practicability and expansibility.

Drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.

FIG. 1 is a schematic structural diagram of a PCR temperature-increasing heating system according to an embodiment of the present invention;

FIG. 2 is a schematic diagram showing the structure of a PCR amplification reaction vessel according to an embodiment of the present invention;

FIG. 3 is a flowchart illustrating steps of a PCR temperature-increasing heating method according to an embodiment of the present invention.

Detailed Description

Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the accompanying drawings are illustrative only for the purpose of explaining the present invention, and are not to be construed as limiting the present invention. The step numbers in the following embodiments are provided only for convenience of illustration, the order between the steps is not limited at all, and the execution order of each step in the embodiments can be adapted according to the understanding of those skilled in the art.

Based on the above background art, most of the conventional PCR heating modules use a semiconductor heating or air heating technique to heat or cool the liquid in the PCR tube by a metal bath or an oil bath. The temperature raising and lowering speed of the module varies from 2.5 degrees (such as Stratagene MX3000P) to 20 degrees (such as LightCycler2.0 of Roche), the temperature raising and lowering speed of the common ABI7500 module is only 2.5 degrees per second, and the temperature raising and lowering speed can directly influence the time consumption and the effect of the PCR reaction. During the heating up, heat is generally generated by the heater, which is generally separated from the pattern carrier to be heated, resulting in heat loss accompanying heat transfer from the heater to the carrier sheet of the sample carrier during heating; in addition, the separation of the heater from the pattern carrier also causes a time delay or lag in the temperature control loop of the heating system or device, and changing the power of the heater to adjust the temperature does not directly and quickly reflect the temperature change of the pattern carrier.

Based on the defects in the prior art, as shown in fig. 1 in a first aspect, the PCR warming and heating system provided by the present application includes a TEC heating module, a heat sink, a heat dissipation fan, and a PCR amplification reaction vessel; the heat radiator is fixedly connected to the first surface of the TEC heating module, a heat radiation fan is arranged on the second surface adjacent to the first surface, and the PCR amplification reaction container is in movable contact with the heat radiator.

Wherein the first surface is a top surface of the TEC heating module after being horizontally placed, and the second surface is a side surface of the TEC heating module; a TEC heating module in the system is used for generating heat; the heat radiator is used for conducting heat to the PCR amplification reaction container; the PCR amplification reaction container is used for heating the PCR amplification reagent by the heat obtained by the conduction of the radiator; the cooling fan is used for guiding the cooling airflow of the TEC heating module.

More specifically, the TEC heating module in the system of the embodiment is mainly used for rapid temperature change; the radiator and the cooling fan are used for accelerating the temperature change of the TEC heating module; the PCR amplification reaction container positioned on the upper part of the TEC heating module changes among different temperatures under the condition that the TEC heating module rapidly rises and falls, the solution in the reaction container can meet the requirement of gene amplification on temperature change, and the reaction container can quickly rise and fall along with the TEC heating module due to the direct contact of the reaction container and the TEC heating module. The invention solves the problems of heat loss in the heat transfer process and inaccuracy of temperature control of the traditional PCR heating device. In addition, the reaction container for placing the PCR amplification reagent in the system of the embodiment, for example, a centrifuge tube for storing the PCR amplification reagent, can be made of plastic, glass, silica gel, silicon wafer, metal and the like, and can be freely selected according to different heating requirements, for example, the requirement condition and rate for temperature control. In the embodiment, the two openings of the PCR amplification reaction vessel have a plurality of sealing modes. The liquid can be prevented from volatilizing by sealing with an oil seal or silica gel and the like, the middle of the oil seal is a reagent, and two ends of the oil seal are blocked by oil seals, and the oil seal can be repeatedly used; or silica gel or other solidifiable glue is selected for plugging as the liquid on the plug, and the liquid can be prevented from volatilizing to the maximum extent.

In some optional embodiments, the PCR warming heating system further comprises a temperature sensor disposed on a surface of the PCR amplification reaction vessel.

Specifically, the TEC heating module directly contacts the temperature sensor, so that real-time monitoring can be realized. The temperature sensor selected in the embodiment is very thin, the conduction effect on the temperature can be ignored, the heat loss is avoided, and the TEC heating module is equivalent to a reaction container directly acting on the temperature sensor.

In some alternative embodiments, the PCR warming heating system may be provided with two heat dissipation fans: the first cooling fan is arranged on a second surface adjacent to the first surface, and the second cooling fan is arranged on a third surface far away from the second surface.

The second surface and the third surface are both side surfaces of the TEC heating module when placed horizontally, and the second surface and the third surface are two side surfaces opposite to each other (or parallel to each other). The heat radiation fans are arranged on the two opposite side surfaces respectively, so that the flow guiding of the heat radiation air flow of the TEC heating module can be accelerated, and the temperature of the TEC heating module can be controlled more quickly.

In some alternative embodiments, as shown in fig. 2, in the PCR warming system, at least one groove is formed on the surface of the PCR amplification reaction vessel away from the heat sink, the grooves are parallel to each other, and the groove is used for placing a centrifuge tube or a chip-type flow channel.

Specifically, there are multiple grooves on the upper surface of the PCR amplification reaction container in parallel, and centrifuge tubes or chip-type flow channels can be placed in the grooves, so that the reagents can be replaced more conveniently, and the PCR amplification reaction container can be used repeatedly and has a long service life. A plurality of groups of different reagents can be simultaneously put into the PCR amplification reaction vessel without mutual interference, thereby effectively improving the accuracy of parallel experiments.

In addition, in some embodiments, the groove of the PCR amplification reaction vessel may further include a communication groove for storing the mixed PCR amplification reagents; the TEC heating module is also connected with an external air pump, and the external air pump is used for controlling the PCR amplification rate.

Specifically, the arrangement mode of the groove or the pipeline in the PCR amplification reaction container in the embodiment system is not fixed, and the application of the system is greatly expanded. For example, two pipelines are opened, and different reagents are respectively put into the two pipelines, so that the related mixing experiment can be carried out. In addition, the system of the embodiment has the advantages that the reagent dosage is small in the heating process, the TEC can control the temperature, the speed can be controlled by controlling the air pressure through the external air pump, and the derivative devices of the system of the embodiment are extremely abundant.

In some optional embodiments, the TEC heating module comprises a hot side and a cold side; the surface of the hot end close to the cold end is provided with a first metal conductor, the surface of the cold end close to the hot end is provided with a second metal conductor, and a plurality of semiconductor elements are arranged between the first metal conductor and the second metal conductor; the hot end and the cold end are ceramic electrodes.

Specifically, the semiconductor cooling/heating device (TEC) in the embodiment is made by using the peltier effect of a semiconductor material; the peltier effect is a phenomenon in which when a direct current passes through a couple composed of two semiconductor materials, one end absorbs heat and the other end releases heat. In the embodiment, heavily doped N-type and P-type bismuth telluride are mainly used as semiconductor materials of TEC, and bismuth telluride elements are electrically connected in series and generate heat in parallel. The TEC heating module comprises a number of P-type and N-type pairs (sets) connected together by electrodes and sandwiched between two ceramic electrodes; when current flows through the TEC, heat generated by the current is transferred from one side of the TEC to the other, creating hot and cold sides on the TEC to achieve an increase or decrease in temperature.

In some alternative embodiments, the PCR amplification reaction vessels in the system are filled with a solution for storing heat and/or for heat transfer.

Specifically, the solution with higher specific heat capacity is filled in the PCR amplification reaction container, so that the heat absorption and heat dissipation capacity of the PCR amplification reaction container is improved, and the requirement of gene amplification on temperature change is met.

In a second aspect, in addition to the system of the first aspect, the present invention provides a PCR warming-up and heating apparatus having any one of the PCR warming-up and heating systems of the first aspect mounted thereon.

In a third aspect, as shown in fig. 3, the present application provides a PCR temperature-increasing heating method based on the system in the first aspect, where the method includes steps S100-S300:

s100, controlling the TEC heating module to heat to generate heat;

s200, conducting heat to the PCR amplification reaction container through a radiator;

s300, heating the PCR amplification reagent by the heat obtained by conducting the PCR amplification reaction container through a radiator;

specifically, in the embodiment, the reagent for PCR amplification is poured into a container such as a centrifuge tube, the container is placed in the PCR amplification reaction container, and then the TEC heating module is controlled to heat, as the temperature of the TEC heating module rises, the heat generated by the TEC heating module is absorbed by the heat sink and further conducted to the PCR amplification reaction container, and the PCR amplification reaction container obtains the heat conducted by the heat sink to heat the reagent for PCR amplification therein. The PCR amplification reaction container positioned at the upper part in the system changes among different temperatures under the condition that the TEC heating module rapidly rises and falls, the solution in the PCR amplification reaction container can meet the requirement of gene amplification on temperature change, and the reaction container can quickly rise and fall along with the TEC heating module due to the direct contact of the reaction container and the TEC heating module.

It can be understood that, since the TEC heating module in the embodiment can perform heating or cooling, the embodiment can also perform cooling on the PCR amplification reagent by heat conduction.

In some alternative embodiments, the PCR warming heating method may further include the steps of:

s400, guiding the heat dissipation airflow of the TEC heating module through a heat dissipation fan.

Specifically, the embodiment dissipates heat through the heat dissipating fan TEC heating module, so that the TEC heating module can reach the target temperature more quickly in the process of temperature regulation and control, especially in the process of temperature reduction.

From the above specific implementation process, it can be concluded that the technical solution provided by the present invention has the following advantages or advantages compared to the prior art:

1. the technical scheme of the application has scientific overall structure design and convenient production, installation and use. The PCR rapid heating system provided by the embodiment mainly solves the problems of rough work, poor precision, complex structure, high production cost and unsuitability for the use of practical micro-fluidic automatic detection equipment of the existing PCR heating device through the installation layout of structural components such as a TEC heating module, a radiator, a cooling fan, a temperature sensor, a reaction container for PCR amplification and the like.

2. According to the technical scheme, the integrated temperature control device is simple in overall structure, wide in application range, capable of being integrated, installed and used, low in production cost, high in temperature control precision and capable of being applied to PCR rapid temperature control device equipment.

In alternative embodiments, the functions/acts noted in the block diagrams may occur out of the order noted in the operational illustrations. For example, two blocks shown in succession may, in fact, be executed substantially concurrently, or the blocks may sometimes be executed in the reverse order, depending upon the functionality/acts involved. Furthermore, the embodiments presented and described in the flow charts of the present invention are provided by way of example in order to provide a more thorough understanding of the technology. The disclosed methods are not limited to the operations and logic flows presented herein. Alternative embodiments are contemplated in which the order of various operations is changed and in which sub-operations described as part of larger operations are performed independently.

Furthermore, although the present invention is described in the context of functional modules, it should be understood that, unless otherwise stated to the contrary, one or more of the functions and/or features may be integrated in a single physical device and/or software module, or one or more of the functions and/or features may be implemented in a separate physical device or software module. It will also be appreciated that a detailed discussion of the actual implementation of each module is not necessary for an understanding of the present invention. Rather, the actual implementation of the various functional modules in the apparatus disclosed herein will be understood within the ordinary skill of an engineer, given the nature, function, and internal relationship of the modules. Accordingly, those skilled in the art can, using ordinary skill, practice the invention as set forth in the claims without undue experimentation. It is also to be understood that the specific concepts disclosed are merely illustrative of and not intended to limit the scope of the invention, which is defined by the appended claims and their full scope of equivalents.

The logic and/or steps represented in the flowcharts or otherwise described herein, e.g., an ordered listing of executable instructions that can be considered to implement logical functions, can be embodied in any computer-readable medium for use by or in connection with an instruction execution system, apparatus, or device, such as a computer-based system, processor-containing system, or other system that can fetch the instructions from the instruction execution system, apparatus, or device and execute the instructions.

In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

While embodiments of the invention have been shown and described, it will be understood by those of ordinary skill in the art that: various changes, modifications, substitutions and alterations can be made to the embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the claims and their equivalents.

While the preferred embodiments of the present invention have been illustrated and described, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

Claims (10)

1. A PCR temperature-rising heating system is characterized by comprising a TEC heating module, a radiator, a cooling fan and a PCR amplification reaction container;

the radiator is fixedly connected to a first surface of the TEC heating module, the radiator fan is arranged on a second surface adjacent to the first surface, and the PCR amplification reaction container is in movable contact with the radiator;

the TEC heating module is used for generating heat; the heat radiator is used for conducting the heat to the PCR amplification reaction vessel; the PCR amplification reaction container is used for heating the PCR amplification reagent by the heat obtained by conduction of the radiator; the heat dissipation fan is used for guiding the heat dissipation airflow of the TEC heating module.

2. The PCR warming and heating system according to claim 1, further comprising a temperature sensor disposed on a surface of the PCR amplification reaction vessel.

3. The PCR thermal system according to claim 1, wherein the heat dissipation fan comprises a first heat dissipation fan and a second heat dissipation fan; the first heat dissipation fan is arranged on a second surface adjacent to the first surface, and the second heat dissipation fan is arranged on a third surface far away from the second surface.

4. The PCR temperature-rising heating system of claim 1, wherein at least one groove is formed on the surface of the PCR amplification reaction vessel far away from the heat radiator, the grooves are parallel to each other, and the grooves are used for placing centrifuge tubes or chip-type flow channels.

5. The PCR thermal heating system according to claim 1, wherein the TEC heating module comprises a first ceramic electrode and a second ceramic electrode; the first ceramic electrode is a hot end, and the first ceramic electrode is a cold end; the surface of the hot end close to the cold end is provided with a first metal conductor, the surface of the cold end close to the hot end is provided with a second metal conductor, and a plurality of semiconductor elements are arranged between the first metal conductor and the second metal conductor.

6. The PCR warming and heating system of claim 4, wherein the grooves comprise a communication groove for storing the PCR amplification reagents after mixing; the TEC heating module is also connected with an external air pump, and the external air pump is used for controlling the PCR amplification rate.

7. A PCR warming and heating system according to any one of claims 1 to 6, wherein the PCR amplification reaction vessel is filled with a solution for storing heat and/or transferring heat.

8. An apparatus for PCR temperature-increasing heating, comprising a PCR temperature-increasing heating system according to any one of claims 1 to 7.

9. A PCR temperature-increasing heating method applied to a PCR temperature-increasing heating system according to any one of claims 1 to 7, comprising the steps of:

controlling the TEC heating module to heat to generate heat;

and conducting the heat to the PCR amplification reaction container through the heat radiator, so that the PCR amplification reaction container conducts the obtained heat to heat the PCR amplification reagent through the heat radiator.

10. The PCR temperature-rising heating method according to claim 9, further comprising the steps of:

and guiding the heat dissipation airflow of the TEC heating module by a heat dissipation fan.

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