A carrier, PCR detection system and PCR amplification method

Abstract

The present application relates to a carrier, a PCR detection system and a PCR amplification method, the carrier comprising a cavity wall, the cavity wall enclosing a reaction cavity for accommodating a reaction solution, at least part of the cavity wall being capable of being heated by an electromagnetic field. The PCR detection system comprises an electromagnetic heating mechanism and a cooling mechanism, the electromagnetic heating mechanism being used for heating the carrier, and the cooling mechanism being used for cooling the carrier. The PCR amplification method uses the carrier and the PCR detection system to perform a PCR amplification method, the PCR amplification method comprising: continuously flowing a cooling liquid in the cooling mechanism to continuously cool the carrier; and the electromagnetic heating mechanism generating a magnetic field and heating the carrier by the magnetic field.

Description

Technical Field

[0001] This invention relates to the field of in vitro diagnostic technology, and in particular to a vector, a PCR detection system, and a PCR amplification method. Background Technology

[0002] PCR (Polymerase Chain Reaction) is a molecular biology experimental method for the in vitro enzymatic synthesis of specific DNA fragments. It mainly consists of three repeated thermal cycles: high-temperature denaturation, low-temperature annealing, and optimal-temperature extension. Before the PCR reaction, the reaction solution needs to be placed in a carrier medium. This reaction solution comprises samples such as throat or nasal swabs and reagents used for the PCR reaction. During the PCR reaction, the reaction solution is heated or cooled by the temperature module of the PCR instrument, thus cycling the reaction solution through the high-temperature denaturation, low-temperature annealing, and optimal-temperature extension stages.

[0003] Existing technologies have long testing times and slow heating and cooling rates, mainly due to factors such as large heater heat capacity, high thermal resistance between the heater and the carrier, and slow thermal equilibrium of the carrier.

[0004] Existing technology uses contact heating, which easily produces condensation during the heating and cooling process. This condensation can cause short circuits and reduce the reliability of the equipment.

[0005] Existing temperature sensing technologies directly detect the temperature of the reaction liquid or carrier, which can easily cause corrosion of the temperature sensor and affect the accuracy of the detection.

[0006] Existing technologies often use fluorescence detection to determine the content of analytes. While existing technologies also disclose methods for determining the temperature of solid objects using fluorescence detection, when this temperature detection method is applied to detect the content of components detected by fluorescence, interference between the fluorescence used for temperature detection and the fluorescence used for component content detection cannot be avoided, leading to poor detection accuracy. Furthermore, existing methods for temperature detection using fluorescence have low sensitivity. Summary of the Invention

[0007] One object of the present invention is to provide a carrier to at least solve one of the above-mentioned technical problems.

[0008] To achieve the above objectives, a first aspect of the present invention provides a carrier including a cavity wall that surrounds a reaction cavity for containing a reaction liquid, wherein at least a portion of the cavity wall is heatable by an electromagnetic field.

[0009] Optionally, the carrier further includes a temperature sensor for reflecting the temperature of the reaction solution.

[0010] Optionally, the temperature sensing unit includes a first fluorescent thermosensitive unit disposed on the outer side of the cavity wall. The first fluorescent thermosensitive unit includes a fluorescent thermosensitive material and can reflect the temperature of the reaction solution.

[0011] Optionally, a light-blocking part is also provided between the first fluorescent temperature-sensitive part and the cavity wall.

[0012] Optionally, the cavity wall includes a first wall and a second wall disposed opposite to each other, and a side wall disposed between the first wall and the second wall, the temperature sensing part is disposed outside the first wall, and at least part of the second wall or at least part of the side wall is made of a transparent material.

[0013] Optionally, at least a portion of the cavity wall is formed of a heat-conducting sheet that can be heated by an electromagnetic field.

[0014] Optionally, the heat-conducting sheet is a metal sheet.

[0015] Optionally, the thickness of the heat-conducting sheet is 10-100 μm.

[0016] Optionally, at least part of the cavity wall is made of a transparent material.

[0017] Optionally, the transparent material is polydimethylsiloxane, polypropylene, plexiglass, or polycarbonate.

[0018] Optionally, the reaction chamber and / or the carrier have a flat structure.

[0019] Optionally, the flat structure refers to a reaction chamber or the carrier having a dimension perpendicular to its thickness direction that is greater than its thickness direction dimension.

[0020] Optionally, the ratio of the dimension of the reaction chamber or the carrier perpendicular to its thickness direction to its dimension in the thickness direction is greater than 5:1.

[0021] Optionally, the size ratio is 50:1 to 100:1.

[0022] Optionally, the electromagnetic heating mechanism is used to heat the carrier, and the cooling mechanism is used to cool the carrier.

[0023] Another object of the present invention is to provide a PCR detection system to at least solve one of the above-mentioned technical problems.

[0024] To achieve this objective, the second aspect of the present invention adopts the following technical solution:

[0025] A PCR detection system includes an electromagnetic heating mechanism and a cooling mechanism, wherein the electromagnetic heating mechanism is used to heat the aforementioned carrier, and the cooling mechanism is used to cool the aforementioned carrier.

[0026] Optionally, the PCR detection system further includes a temperature detection mechanism for detecting the temperature of the reaction solution within the carrier.

[0027] Optionally, the temperature detection mechanism is a first temperature detection mechanism, which detects the temperature of the reaction solution by measuring the fluorescence intensity at the temperature sensing part of the detection carrier.

[0028] Optionally, the first temperature detection mechanism includes:

[0029] The first fluorescent transceiver assembly is used to emit excitation light of a preset intensity to the temperature sensing unit and to receive and detect the intensity of the excitation light at the temperature sensing unit.

[0030] A first controller is electrically connected to the first fluorescent transceiver assembly and is used to convert the light intensity signal detected by the first fluorescent transceiver assembly into a temperature signal.

[0031] Optionally, the first fluorescent transmitter / receiver assembly includes:

[0032] A first light source is used to emit the excitation light of a preset intensity;

[0033] The first incident channel is connected to the first light source and is used to transmit the excitation light of the first light source to the temperature sensing unit.

[0034] A first receiving channel and a first detector, wherein the first receiving channel is used to transmit the excitation light at the temperature sensing unit to the first detector, and the first detector is capable of detecting the intensity of the excitation light transmitted by the first receiving channel.

[0035] Optionally, the temperature detection mechanism is a second temperature detection mechanism, which is used to detect the temperature of the reaction liquid inside the carrier.

[0036] Optionally, the second temperature detection mechanism includes:

[0037] The second fluorescent thermosensitive part includes a fluorescent thermosensitive material, and the second fluorescent thermosensitive part can be located in the reaction liquid within the reaction chamber of the carrier;

[0038] The second fluorescent transceiver assembly is used to emit excitation light of a preset intensity to the second fluorescent thermosensitive part and to receive and detect the intensity of the excitation light at the second fluorescent thermosensitive part.

[0039] The second controller is electrically connected to the second fluorescent transceiver assembly and is used to convert the light intensity signal detected by the second fluorescent transceiver assembly into a temperature signal.

[0040] Optionally, the second fluorescent transmitter assembly includes:

[0041] The second incident channel is used to transmit the excitation light to the second fluorescence temperature-sensitive part;

[0042] The second receiving channel is used to transmit the excitation light at the second fluorescent temperature-sensitive part;

[0043] The second exit end of the second incident channel, the second incident end of the second receiving channel, and the second fluorescence thermosensitive part can all be located in the reaction liquid of the reaction chamber.

[0044] Optionally, the second temperature detection mechanism further includes a light-blocking covering, wherein at least the second exit end of the second incident channel, at least the second incident end of the second receiving channel, and the second fluorescent temperature-sensitive part are all disposed within the light-blocking covering.

[0045] Optionally, the second fluorescent temperature-sensitive portion includes a fluorescent temperature-sensitive material coated on the second exit end of the second incident channel; and / or, the second fluorescent temperature-sensitive portion is connected to the second exit end of the second incident channel and extends from the second incident channel.

[0046] Optionally, the cooling mechanism includes a container in which coolant is continuously circulated, and the carrier can be inserted into the coolant.

[0047] Optionally, the electromagnetic heating mechanism includes a coil for carrying current.

[0048] Optionally, the coil is disposed on the upper or lower side of the container, or the coil is sleeved on the outside of the container, or the coil is disposed inside the container, and / or

[0049] The PCR detection system also includes a fluorescence detection device, which is used to detect the content of the analyte in the reaction solution.

[0050] Optionally, the fluorescence detection device is located outside the container.

[0051] Optionally, the fluorescence detection device is located on one side of the axial direction or the radial direction of the coil.

[0052] Another object of the present invention is to provide a PCR amplification method to at least solve one of the above-mentioned technical problems.

[0053] To achieve this objective, the third aspect of the present invention adopts the following technical solution:

[0054] A PCR amplification method, comprising performing the PCR amplification method using the vector and the PCR detection system described above, the PCR amplification method including:

[0055] The coolant flows continuously within the cooling system to continuously cool the medium.

[0056] The electromagnetic heating mechanism generates a magnetic field, which heats the carrier.

[0057] Optionally, by adjusting the power of the electromagnetic heating mechanism, and / or

[0058] The temperature of the carrier is adjusted by regulating the temperature and / or flow rate of the coolant within the cooling mechanism; and / or

[0059] The coolant continuously flows within the cooling mechanism to continuously cool the carrier, including:

[0060] The carrier is in direct contact with the coolant.

[0061] Optionally, the PCR amplification method further includes:

[0062] The temperature of the reaction solution is detected by using a fluorescent thermosensitive material and the intensity of excitation light.

[0063] Optionally, the detection of the temperature of the reaction solution by means of the fluorescent thermosensitive material and the intensity of the excitation light includes:

[0064] The fluorescent thermosensitive material continuously emits excitation light;

[0065] Collect the excitation light from the fluorescent temperature-sensitive material.

[0066] The intensity of the collected excitation light is obtained, and the temperature of the reaction solution is obtained from the intensity of the collected excitation light.

[0067] As can be seen from the above, the technical solution provided by this invention allows the cavity wall to be heated by an electromagnetic field. Therefore, the carrier can be heated non-contactly using an electromagnetic field. Since the electromagnetic heating mechanism that generates the electromagnetic field does not contact the carrier, even if condensation occurs on the carrier during heating or cooling, it will not cause a short circuit in the electromagnetic heating mechanism's circuit, thus improving the reliability of the equipment. The electromagnetic heating mechanism that generates the electromagnetic field can heat the reaction liquid inside the carrier without contact, thus saving the time required for heat conduction between the carrier and the electromagnetic heating mechanism, improving heating efficiency, and accelerating the PCR reaction process and efficiency. This solves the problems of long detection times and slow heating / cooling rates caused by the large heat capacity and high thermal resistance between the heater and the carrier in traditional methods.

[0068] The carrier is formed directly from a heat-conducting sheet or a heat-conducting sheet and a transparent material, which reduces the heat capacity of the carrier, allows the reaction liquid to heat up quickly, and results in low carrier cost and a simple process.

[0069] The carrier may further include a first fluorescent thermosensitive element, and a first temperature detection mechanism detects the temperature of the reaction solution by detecting the fluorescence intensity at the first fluorescent thermosensitive element of the carrier. Because the reaction chamber is very thin, the temperature of the reaction solution can be quickly and uniformly distributed throughout. Simultaneously, heat transfer between the chamber wall and the reaction solution is rapid; therefore, the temperature of the chamber wall can be considered to be the same as the temperature of the reaction solution. The first fluorescent thermosensitive element is located on the outside of the carrier and on the chamber wall; therefore, heat from the chamber wall can be quickly conducted to the second fluorescent thermosensitive element, enabling the second fluorescent thermosensitive element to reflect the temperature of the reaction solution in real time and accurately.

[0070] The cavity wall has a certain light-blocking function. When the first fluorescent temperature-sensitive part is directly placed on the outside of the cavity wall, the cavity wall can prevent the fluorescence used for temperature detection from entering the reaction cavity, thus avoiding interference with the detection results of the fluorescence detection device and ensuring the accuracy of PCR detection.

[0071] A light-blocking section is also provided between the first fluorescence temperature-sensitive part and the cavity wall. When light leaks from the cavity wall, the light-blocking section can completely prevent the fluorescence used for temperature detection from entering the reaction cavity, thus completely avoiding interference from the fluorescence used for temperature detection on the detection results of the fluorescence detection device and ensuring the accuracy of PCR detection.

[0072] The reaction chamber and / or carrier have a flat structure, which makes the reaction liquid inside the reaction chamber very thin and the center of the reaction liquid is very close to the surface of the liquid. When the carrier is heated / cooled, the temperature of the reaction liquid can reach uniformity in a very short time, which improves the thermal equilibrium rate of the carrier and greatly improves the heating and cooling rate of the reaction liquid and the detection efficiency.

[0073] The PCR detection system includes an electromagnetic heating mechanism and a cooling mechanism. The electromagnetic heating mechanism heats the carrier, and the cooling mechanism cools the carrier. The electromagnetic heating mechanism generates an alternating magnetic field, inducing eddy currents in the heat-conducting sheet through magnetic field eddy current heating, thereby raising the temperature of the carrier and subsequently the reaction solution within it. The electromagnetic heating mechanism heats the carrier through a non-contact electromagnetic field; since it does not come into contact with the carrier, even if condensation occurs during the heating and cooling process, it will not cause a short circuit in the electromagnetic heating mechanism's circuitry, thus improving the reliability of the equipment. Because the electromagnetic heating mechanism heats the reaction solution within the carrier without direct contact, it saves the time required for heat conduction between the carrier and the traditional heater, as well as the time required for the heater itself to heat up. This results in rapid heating of the carrier and reaction solution, improving heating efficiency and accelerating the PCR reaction process and efficiency. It solves the problems of long detection times and slow heating and cooling rates caused by the large heat capacity of the heater and the high thermal resistance between the heater and the carrier.

[0074] The cooling mechanism is used to cool the carrier, thereby lowering the temperature of the reaction solution inside the carrier. The electromagnetic heating mechanism and the cooling mechanism keep the reaction solution at the denaturation temperature, extension temperature and annealing temperature of the PCR reaction process, respectively, and maintain them for a period of time.

[0075] When the reaction solution needs cooling, the power of the electromagnetic heating mechanism can be reduced, such as by increasing the duty cycle or decreasing the current, and / or increasing the flow rate of the coolant. When the power of the electromagnetic heating mechanism changes, the eddy currents within the carrier change instantaneously, and the heat generated by the carrier itself also changes accordingly. Therefore, the PCR detection system can more easily and quickly adjust the temperature of the carrier and the reaction solution. The cooling mechanism includes a container with a continuous flow of coolant. The carrier can be inserted into the coolant, meaning the coolant is in direct contact with the carrier. The coolant flows continuously during the PCR reaction, so the cooling mechanism is always ready to cool the carrier, allowing for rapid cooling when needed.

[0076] The first fluorescent transducer (or the second fluorescent transducer, i.e., the second light source) emits monochromatic excitation light with a wavelength of λ0. When this excitation light illuminates the first fluorescent thermosensitive part (or the second fluorescent thermosensitive part), the fluorescent molecules with wavelength λ0 will generate light of a different wavelength as the temperature of the first fluorescent thermosensitive part (or the second fluorescent thermosensitive part) changes. This results in a decrease in the intensity of the excitation light with wavelength λ0. The first receiving channel (or the second receiving channel) collects the intensity I of the weakened excitation light to obtain the temperature. The PCR detection system is unaffected by the monochromaticity (i.e., the wavelength distribution of the monochromatic light) of the excitation light emitted by the first fluorescent transducer (or the second fluorescent transducer, i.e., the second light source), which improves detection accuracy and sensitivity, and ensures accurate temperature control.

[0077] The second exit end of the second incident channel, the second incident end of the second receiving channel, and the second fluorescence thermosensitive part can all be located in the reaction solution of the reaction chamber. The fact that the second fluorescence thermosensitive part is located in the reaction solution of the reaction chamber ensures that the temperature of the second fluorescence thermosensitive part is consistent with that of the reaction solution, thereby improving the temperature detection accuracy and thus improving the PCR detection accuracy. The fact that the second exit end of the second incident channel and the second incident end of the second receiving channel are located in the reaction solution of the reaction chamber ensures that the distance between them and the second fluorescence thermosensitive part is very small, thereby improving the accuracy of fluorescence intensity detection and thus ensuring the accuracy of temperature detection.

[0078] The second temperature detection mechanism also includes a light-shielding enclosure, in which at least the second exit end of the second incident channel, at least the second incident end of the second receiving channel, and the second fluorescence temperature-sensitive part are all disposed. In this embodiment, the light used for temperature detection is distributed within the light-shielding enclosure and will not leak outside the light-shielding enclosure, thus avoiding interference from the light of the temperature detection mechanism on the detection results of the fluorescence detection device and ensuring the accuracy of PCR detection.

[0079] PCR amplification methods include:

[0080] The coolant flows continuously within the cooling system to continuously cool the medium.

[0081] The electromagnetic heating mechanism generates a magnetic field, which heats the carrier.

[0082] The electromagnetic heating mechanism generates a magnetic field, causing the carrier to heat up rapidly, which in turn raises the temperature of the reaction solution inside the carrier. The cooling mechanism cools the carrier, thereby lowering the temperature of the reaction solution inside. The electromagnetic heating and cooling mechanisms maintain the reaction solution at the denaturation, extension, and annealing temperatures required in the PCR reaction process for a period of time. The coolant in the cooling mechanism flows continuously, ready to provide cooling at any time for rapid cooling.

[0083] The electromagnetic heating mechanism heats the carrier through a non-contact electromagnetic field. Since the mechanism does not come into contact with the carrier, even if condensation occurs on the carrier during heating or cooling, it will not cause a short circuit in the electromagnetic heating mechanism, thus improving the reliability of the equipment. Furthermore, because the electromagnetic heating mechanism can heat the reaction liquid within the carrier without direct contact, it saves the time required for heat conduction between the carrier and the electromagnetic heating mechanism, achieving rapid heating and accelerating the PCR reaction process and efficiency. This solves the problems of long detection times and slow heating / cooling rates caused by the large heat capacity of the heater and the high thermal resistance between the heater and the carrier. Attached Figure Description

[0084] Figure 1 This is a schematic diagram of the structure of the carrier provided in an embodiment of the present invention;

[0085] Figure 2a This is a schematic diagram of the structure of a PCR detection system provided in an embodiment of the present invention;

[0086] Figure 2b This is the temperature rise curve of the reaction solution when the PCR detection system provided in this embodiment is used to heat the reaction solution in the vector;

[0087] Figure 3 This is a schematic diagram of another PCR detection system provided in an embodiment of the present invention;

[0088] Figure 4 This is a schematic diagram of the structure connecting the second emission end and the second fluorescence temperature-sensitive part provided in an embodiment of the present invention;

[0089] Figure 5 This is the curve showing the relationship between the excitation light intensity (I) and temperature (T) collected in the embodiments of the present invention;

[0090] Figure 6a This is a schematic diagram of the structure of another carrier provided in an embodiment of the present invention;

[0091] Figure 6b This is a schematic diagram of the structure of another carrier provided in an embodiment of the present invention;

[0092] Figure 7 This is a schematic diagram of the structure of another PCR detection system provided in an embodiment of the present invention;

[0093] Figure 8 This is a schematic diagram of another PCR detection system provided in an embodiment of the present invention.

[0094] In the picture:

[0095] 1. Carrier; 11. Cavity wall; 111. First wall; 112. Second wall; 113. Side wall; 114. Transparent material; 115. Thermally conductive sheet; 12. Reaction chamber; 13. First fluorescent temperature-sensitive part; 14. Light-blocking part;

[0096] 2. Coil;

[0097] 3. First temperature detection mechanism; 31. First incident channel; 32. First receiving channel; 33. First fluorescence transceiver assembly; 34. First detector;

[0098] 4. Second temperature detection mechanism; 41. Second fluorescence temperature-sensitive part; 42. Second incident channel; 421. Second exit end; 43. Second receiving channel; 44. Light-blocking covering part; 45. Second fluorescence receiving and transmitting assembly; 46. Second detector;

[0099] 5. Container;

[0100] 6. Fluorescence detection device. Detailed Implementation

[0101] The technical solution of the present invention will be further described below with reference to the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and not intended to limit it. Furthermore, it should be noted that, for ease of description, only the parts relevant to the present invention are shown in the accompanying drawings, not all of them.

[0102] This invention defines certain directional terms. Unless otherwise stated, the directional terms used, such as "up," "down," "left," "right," "inner," and "outer," are used for ease of understanding and therefore do not constitute a limitation on the scope of protection of this invention.

[0103] In this invention, unless otherwise explicitly specified and limited, "above" or "below" the second feature can include direct contact between the first and second features, or contact between the first and second features through another feature between them. Furthermore, "above," "over," and "on top" of the second feature includes the first feature directly above or diagonally above the second feature, or simply indicates that the first feature is at a higher horizontal level than the second feature. "Below," "below," and "under" the second feature includes the first feature directly below or diagonally below the second feature, or simply indicates that the first feature is at a lower horizontal level than the second feature.

[0104] In the description of this invention, unless otherwise explicitly specified and limited, the terms "connected," "linked," and "fixed" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances.

[0105] Example 1

[0106] This embodiment provides a vector 1 for use in PCR reactions and PCR detection, but is not limited thereto.

[0107] like Figure 1 As shown, the carrier 1 provided in this embodiment includes a cavity wall 11, which forms a reaction cavity 12 for containing the reaction liquid, and at least a portion of the cavity wall 11 can be heated by an electromagnetic field.

[0108] The cavity wall 11 provided in this embodiment can be heated by an electromagnetic field. Therefore, the carrier 1 can be heated by a non-contact electromagnetic field. Since the electromagnetic heating mechanism that generates the electromagnetic field does not contact the carrier 1, even if condensation occurs in the carrier 1 during the heating and cooling process, the circuit of the electromagnetic heating mechanism will not be short-circuited, thereby improving the reliability of the device. The electromagnetic heating mechanism that generates the electromagnetic field can heat the reaction liquid inside the carrier 1 without contact with the carrier 1. Therefore, when heating the carrier 1, the time required for heat conduction between the carrier 1 and the electromagnetic heating mechanism is saved, improving heating efficiency and accelerating the PCR reaction process and PCR reaction efficiency. This solves the problems of long detection time and slow heating and cooling rates caused by the large heat capacity of traditional heaters and the large thermal resistance between the heater and the carrier 1.

[0109] Optionally, at least a portion of the cavity wall 11 of the carrier 1 is formed by a heat-conducting sheet 115, which can be heated by an electromagnetic field. The heat-conducting sheet 115 is thin and has thermal conductivity; therefore, when the reaction liquid needs to be cooled, the heat of the reaction liquid can be quickly transferred to the outside of the heat-conducting sheet 115. The heat-conducting sheet 115 can also be referred to as a thermally conductive film. Optionally, the thickness of the heat-conducting sheet 115 can be 10-100 μm. While ensuring the strength of the carrier 1, it can improve the thermal conductivity of the carrier 1, thereby improving detection efficiency and shortening detection time.

[0110] Optionally, the heat-conducting plate 115 is a metal plate, which has both high thermal conductivity and can generate eddy currents in an alternating electromagnetic field, thereby being heated in an alternating magnetic field. Optionally, the metal plate is made of nickel or a nickel alloy.

[0111] Optionally, the cavity wall 11 includes a first wall 111 and a second wall 112 disposed opposite to each other, and a side wall 113 disposed between the first wall 111 and the second wall 112. The side wall 113, the first wall 111, and the second wall 112 can form a reaction cavity 12. It is understood that the side wall 113 is annular, meaning that the side wall 113 extends continuously until both ends are closed. The annular shape can be a circular ring, an elliptical ring, a square ring, or a polygonal ring, etc., to close the space between the first wall 111 and the second wall 112.

[0112] The reaction chamber 12 and / or the carrier 1 have a flat structure, which makes the reaction liquid in the reaction chamber 12 very thin, such as 0.3-1mm. The center of the reaction liquid is very close to the surface of the liquid. When the carrier 1 is heated / cooled, the temperature of the reaction liquid can reach uniformity in a very short time, which improves the thermal equilibrium rate of the carrier 1 and greatly improves the heating and cooling rate of the reaction liquid and the detection efficiency.

[0113] Optionally, when the carrier 1 has a flat structure, it can accommodate a flat-structured reaction chamber 12. Specifically, a flat structure means that the dimension in the thickness direction of the reaction chamber 12 or the carrier 1 is smaller than the dimension perpendicular to the thickness direction. For example, the ratio of the dimension perpendicular to the thickness direction to the dimension in the thickness direction is greater than 5:1. More preferably, the dimension in the thickness direction of the carrier 1 or the reaction chamber 12 is much smaller than the dimension perpendicular to the thickness direction, such as a dimension ratio of 50:1 to 100:1. For example, a dimension ratio of 90:1. For example, the reaction chamber 12 is a cuboid, and the ratio of the length to the thickness of the cuboid can be greater than 5:1, such as 90:1. For example, the dimension in the thickness direction of the reaction chamber 12 can be 0.3-1.0 mm. In this embodiment, the thickness of the reaction chamber 12 is 0.3-0.6 mm, and the width and length of the reaction chamber 12 are approximately 10 mm and 20 mm, respectively. As an example, the reaction chamber 12 can also be a cylindrical structure with a diameter-to-thickness ratio greater than 5:1, such as a thickness of 0.3-1.0 mm and a diameter of 5-20 mm. Of course, the cross-section of the reaction chamber 12 can be polygonal, circular, or elliptical, etc.

[0114] Optionally, at least a portion of the cavity wall 11 is made of a transparent material 114, for example, a portion of the sidewall 113 (such as...) Figure 1 (as shown) or part of the first wall 111 or part of the second wall 112 (as shown) Figure 6a and Figure 6b (As shown) is made of transparent material 114. Optionally, the cavity wall 11, except for the transparent material 114, is made of heat-conducting sheet 115. Excitation light interacts with fluorescent probes / dyes in the reaction solution to generate fluorescence. The fluorescence can pass through the transparent material 114, thereby being detected by the fluorescence detection device 6 to realize the detection of the content of the analyte in the reaction solution.

[0115] Optionally, the transparent material 114 can be polydimethylsiloxane, polypropylene, plexiglass, or polycarbonate. PMMA, PDMS, PP, and PC are optically transparent materials with good biocompatibility, thus meeting the requirements for fluorescence detection and having no impact on the reaction sample.

[0116] The carrier 1 provided in this embodiment is formed directly from a thermally conductive sheet or a thermally conductive sheet and a transparent material 114, which reduces the heat capacity of the carrier 1, resulting in a rapid heating rate of the reaction solution. The carrier 1 is also low in cost and simple in process. For example, flat carrier 1 can be mass-produced using metal sheets (metal films), which is also low in cost and simple in process. Using an ultra-thin carrier 1 (i.e., the thickness dimension of the carrier 1 is much smaller than the dimensions in other directions) reduces the total heat capacity of the carrier 1, increases the thermal equilibrium rate between the reaction solution and the carrier 1, and thus improves the heating and cooling rates.

[0117] like Figure 2aAs shown, this embodiment also provides a PCR detection system, which includes an electromagnetic heating mechanism and a cooling mechanism. The electromagnetic heating mechanism is used to heat the carrier 1, and the cooling mechanism is used to cool the carrier 1. The electromagnetic heating mechanism can generate an alternating magnetic field, and through magnetic field eddy current heating, eddy currents are induced in the heat-conducting plate 115, thereby heating the carrier 1 and thus rapidly heating the reaction solution inside the carrier 1. The electromagnetic heating mechanism heats the carrier 1 through a non-contact electromagnetic field. The electromagnetic heating mechanism does not come into contact with the carrier 1, so even if condensation occurs on the carrier 1 during heating and cooling, it will not cause a short circuit in the circuit of the electromagnetic heating mechanism, thereby improving the reliability of the device. Also, since the electromagnetic heating mechanism can heat the reaction solution inside the carrier 1 without contact with the carrier 1, the non-contact electromagnetic heating mechanism heats the carrier 1, saving the time required for heat conduction between the carrier 1 and the traditional heater, as well as the time required for the heater itself to heat up. The carrier 1 and the reaction solution heat up quickly, improving heating efficiency and accelerating the PCR reaction process and PCR reaction efficiency. This solves the problems of long detection time and slow heating and cooling rates caused by the large heat capacity of the heater and the large thermal resistance between the heater and the carrier 1. Figure 2b As shown in the figure, after experimental verification, when the reaction solution in the vector 1 is heated using the PCR detection system provided in this embodiment, the heating rate of the reaction solution can reach 100℃ / S.

[0118] The cooling mechanism is used to cool the carrier 1, thereby cooling the reaction solution inside the carrier 1. The electromagnetic heating mechanism and the cooling mechanism keep the reaction solution at the denaturation temperature, extension temperature and annealing temperature in the PCR reaction process, respectively, and maintain them for a period of time.

[0119] For example, when the reaction solution needs to be cooled, the power of the electromagnetic heating mechanism can be reduced, such as by increasing the duty cycle or reducing the current, and / or increasing the flow rate of the coolant. When the power of the electromagnetic heating mechanism changes, the eddy current in the carrier 1 changes instantaneously, and the heat generated by the carrier 1 itself also changes. Therefore, the PCR detection system can more easily and quickly adjust the temperature of the carrier 1 and the reaction solution.

[0120] Optionally, the cooling mechanism includes a container 5, in which coolant continuously flows. The carrier 1 can be inserted into the coolant, meaning the coolant directly contacts the carrier 1, eliminating the need for other heat-conducting structures between the carrier and the coolant, thus enabling rapid cooling of the carrier. Preferably, the entire carrier 1 is inserted into the coolant; however, a portion of the carrier 1 can also be inserted. By inserting the carrier 1 into the coolant, the coolant continues to flow during the PCR reaction, ensuring the cooling mechanism is always ready to cool the carrier 1, allowing for rapid cooling when needed.

[0121] Optionally, the electromagnetic heating mechanism includes a coil 2 for carrying current. When an alternating current is passed through the coil 2, a magnetic field is generated. This magnetic field interacts with the carrier 1, generating eddy currents in the carrier 1, causing the carrier 1 to heat up rapidly. The electromagnetic heating mechanism has a simple structural design, is easy to manufacture, and has low cost.

[0122] For example, coil 2 is disposed on the underside of container 5 (e.g., Figure 2a (As shown), or coil 2 is placed on the upper side of container 5, or coil 2 is sleeved on the outside of container 5, with coil 2 located outside container 5, thereby avoiding short circuit of coil 2.

[0123] In other alternative embodiments, the coil 2 may also be disposed within the container 5. Optionally, when the coil 2 is disposed within the container 5, a separate space may be provided within the container 5 to accommodate the coil 2 and to isolate the coolant to prevent short circuits.

[0124] This embodiment of the PCR detection system employs direct contact cooling with the coolant, meaning the coolant directly contacts the carrier 1 for cooling, resulting in rapid cooling and shortening the PCR reaction time. The alternating magnetic field generated by coil 2 induces eddy currents on the heat-conducting plate 115, causing the carrier 1 to heat up itself, achieving non-contact eddy current heating. This eliminates the need for heat conduction between the heater and the carrier 1, resulting in rapid heating and further shortening the PCR reaction time. Non-contact heating provides rapid heating and eliminates the condensation that occurs in traditional contact heating and cooling processes, improving the system's reliability.

[0125] like Figure 2a As shown, optionally, the PCR detection system also includes a fluorescence detection device 6, which is used to detect the content of the analyte in the reaction solution. The fluorescence detection device 6 can be an existing structure, and the structure of the fluorescence detection device 6 will not be described in detail here.

[0126] Optionally, the fluorescence detection device 6 is located outside the container 5 to prevent the coolant from damaging the fluorescence detection device 6.

[0127] like Figure 2a As shown, exemplaryly, the fluorescence detection device 6 is located on one side of the coil 2 along its axial direction to make the PCR detection system structure compact. For example, the fluorescence detection device 6 is disposed on the upper side of the container 5, and at least a portion of the upper end of the sidewall 113 of the carrier 1 is made of a transparent material 114. In an optional embodiment, the sidewall 113 includes a top wall and a bottom wall disposed opposite each other, wherein the top wall is located at the upper end of the carrier 1 and the top is made of a transparent material 114. In other optional embodiments, the fluorescence detection device 6 may be disposed on one side of the coil 2 in a radial direction, thereby reducing interference from the magnetic field on the fluorescence detection device 6.

[0128] Optionally, the PCR detection system may also include a temperature detection mechanism for detecting the temperature of the reaction solution, thereby accurately controlling the operation of the heating and / or cooling mechanisms. Optionally, the temperature detection mechanism detects the temperature by detecting the fluorescence intensity.

[0129] Example 2

[0130] like Figure 3 As shown, the vector 1 and PCR detection system in this second embodiment are basically the same as those in the first embodiment. The difference is that the temperature detection mechanism is a second temperature detection mechanism 4, which is used to detect the temperature of the reaction solution in the vector 1 in order to better control the PCR reaction.

[0131] like Figure 3 and Figure 5 As shown, the second temperature detection mechanism 4 includes a second fluorescent temperature-sensitive part 41, a second fluorescent transducer assembly 45, and a second controller. The second fluorescent temperature-sensitive part 41 comprises a fluorescent temperature-sensitive material and can be located in the reaction liquid within the reaction chamber 12 of the carrier 1. The second fluorescent transducer assembly 45 is used to emit excitation light of a preset intensity I0 to the second fluorescent temperature-sensitive part 41 and to receive and detect the intensity of the excitation light I1 at the second fluorescent temperature-sensitive part 41.

[0132] The second controller is electrically connected to the second fluorescent transducer 45 and is used to convert the light intensity signal detected by the second fluorescent transducer 45 into a temperature signal. The PCR detection system may also include a control mechanism, which detects the real-time temperature of the reaction solution through the second temperature detection mechanism 4 and feeds it back to the control mechanism. The control mechanism recognizes the feedback real-time temperature and adjusts the temperature of the carrier 1 and the reaction solution according to the temperature by controlling the power of the electromagnetic heating mechanism, etc., so that they are respectively at the denaturation temperature, extension temperature and annealing temperature in the PCR reaction process, and maintain them for a period of time, thereby realizing PCR amplification. The fluorescence detection device 6 performs fluorescence detection on the amplified reaction solution.

[0133] Optionally, the fluorescent thermosensitive material can be at least one of Rhodamine B, fluorescein, and anthocyanin; of course, other materials can also be used. This embodiment uses a fluorescent thermosensitive material to detect the temperature of the reaction solution, achieving accurate temperature measurement with high sensitivity.

[0134] like Figure 3 As shown, optionally, the second fluorescence transceiver assembly 45 includes a second incident channel 42 and a second receiving channel 43. The second incident channel 42 is used to transmit excitation light to the second fluorescence temperature-sensitive part 41, and the second receiving channel 43 is used to transmit excitation light at the second fluorescence temperature-sensitive part 41. Optionally, the second incident channel 42 and the second receiving channel 43 are optical fibers.

[0135] like Figure 3 and Figure 5 As shown, the second fluorescent transmitter assembly 45 may further include a second light source connected to the second incident channel 42 to emit excitation light into the second incident channel 42. The second light source preferably emits monochromatic light with a wavelength of λ0.

[0136] The working principle of the second fluorescence detection unit is as follows: Figure 5 As shown, the second fluorescent transducer 45 (i.e., the second light source) emits monochromatic excitation light with a wavelength of λ0. After the excitation light emitted by the second fluorescent transducer 45 irradiates the second fluorescent thermosensitive part 41, the fluorescent molecules with a wavelength of λ0 will generate light of another wavelength as the temperature of the second fluorescent thermosensitive part 41 changes, thereby weakening the intensity of the excitation light with a wavelength of λ0. The second receiving channel 43 collects the intensity I of the weakened excitation light. The relationship between intensity I and temperature T is as follows: Figure 5 As shown, and satisfies the formula: The units for T1 and T2 are K.

[0137] The detection method provided in this embodiment is not affected by the monochromaticity (i.e., wavelength distribution of monochromatic light) of the excitation light emitted by the second fluorescent transducer 45 (i.e., the second light source), which can improve detection accuracy and sensitivity and ensure the accuracy of temperature control.

[0138] The second fluorescent temperature-sensitive portion 41 includes a fluorescent temperature-sensitive material coated on the second emission end 421 of the second incident channel 42; and / or, such as Figure 4 As shown, optionally, the second fluorescence temperature-sensitive unit 41 is connected to the second fluorescence transceiver assembly 45. Specifically, the second fluorescence temperature-sensitive unit 41 is connected to the second emission end 421 of the second incident channel 42 and extends from the second incident channel 42. The arrangement of the second fluorescence temperature-sensitive unit 41 ensures that the excitation light incident on the second incident channel 42 is all irradiated by the second fluorescence temperature-sensitive unit 41, thereby accurately obtaining the intensity of the excitation light emitted by the second incident channel 42 and the excitation light collected by the second receiving channel 43, thus improving the detection accuracy.

[0139] The second exit end 421 of the second incident channel 42, the second incident end of the second receiving channel 43, and the second fluorescence temperature-sensitive part 41 can all be located in the reaction solution of the reaction chamber 12. The second fluorescence temperature-sensitive part 41 being located in the reaction solution of the reaction chamber 12 ensures that the temperature of the second fluorescence temperature-sensitive part 41 is consistent with that of the reaction solution, thereby improving the accuracy of temperature detection and thus improving the accuracy of PCR detection. The second exit end 421 of the second incident channel 42 and the second incident end of the second receiving channel 43 being located in the reaction solution of the reaction chamber 12 ensures that the distance between them and the second fluorescence temperature-sensitive part 41 is very small, thereby improving the accuracy of fluorescence intensity detection and thus ensuring the accuracy of temperature detection.

[0140] Furthermore, the first incident channel 31 and the first receiving channel 32 can be arranged adjacent to each other to further improve detection accuracy.

[0141] Preferably, the second temperature detection mechanism 4 further includes a light-shielding covering part 44, wherein at least the second exit end 421 of the second incident channel 42, at least the second incident end of the second receiving channel 43, and the second fluorescence temperature-sensitive part 41 are all disposed within the light-shielding covering part 44. In this embodiment, the light used for temperature detection is distributed within the light-shielding covering part 44 and will not leak outside the light-shielding covering part 44, which can prevent the light from the temperature detection mechanism from interfering with the detection results of the fluorescence detection device 6 and ensure the accuracy of PCR detection.

[0142] Optionally, the light-blocking covering 44 can be an opaque flexible bag, such as one made of polypropylene (PP) or polycarbonate (PC). The second fluorescent temperature-sensitive part 41 can also be a fluorescent liquid, which is placed inside the light-blocking covering 44. The second exit end 421 of the second incident channel 42 and the second incident end of the second receiving channel 43 can extend into the fluorescent liquid or be located above the fluorescent liquid.

[0143] Optionally, an insertion hole is provided on the side wall 113 of the carrier 1, and the portion of the second temperature detection mechanism 4 located inside the reaction chamber 12 can be inserted into the reaction chamber 12 through the insertion hole. Specifically, the insertion hole can be interference-fitted with the portion of the second temperature detection mechanism 4 inserted into the reaction chamber 12, thereby preventing water vapor from escaping during the PCR reaction. For example, the portion of the second temperature detection mechanism 4 located inside the reaction chamber 12 includes a second exit end 421 of the second incident channel 42, a second incident end of the second receiving channel 43, a second fluorescence temperature-sensitive part 41, and a light-blocking coating part 44.

[0144] Optionally, the second fluorescence transceiver assembly 45 may further include a second detector 46, which is connected to the second receiving channel 43 and is used to detect the intensity of the excitation light transmitted by the second receiving channel 43. The fluorescence detection device 6 includes a fluorescence detection detector, which is used to detect the intensity of fluorescence. In this embodiment, the specific locations of the fluorescence detection detector and the second detector 46 are not limited, as long as both the first temperature detection mechanism and the fluorescence detection device 6 can detect the corresponding fluorescence intensity.

[0145] The detector can include silicon photomultiplier tubes, photonic detectors, or photomultiplier tubes. Detectors using silicon photomultiplier tubes, photonic detectors, or photomultiplier tubes have high sensitivity, enabling ultrafast and high-sensitivity detection of fluorescence signals at the millisecond level.

[0146] Example 3

[0147] like Figures 6a-7 As shown, the vector 1 and PCR detection system in this third embodiment are basically the same as those in the first embodiment. The difference is that the vector 1 is further provided with a structure for detecting temperature. The temperature detection mechanism is a first temperature detection mechanism 3, which is used to detect the temperature of the reaction solution inside the vector 1 to better control the PCR reaction. Of course, the PCR detection system may also include both the first temperature detection mechanism 3 and the second temperature detection mechanism 4 from the second embodiment.

[0148] The carrier 1 may also include a temperature sensing unit, which is used to reflect the temperature of the reaction liquid, thereby facilitating the first temperature detection mechanism 3 to detect the temperature of the reaction liquid.

[0149] like Figure 6a As shown, the temperature sensing unit includes a first fluorescent temperature-sensitive part 13. The first temperature detection mechanism 3 detects the temperature of the reaction liquid by measuring the fluorescence intensity at the first fluorescent temperature-sensitive part 13 on the detection carrier 1. Optionally, the first fluorescent temperature-sensitive part 13 is disposed on the outside of the cavity wall 11. The first fluorescent temperature-sensitive part 13 includes a fluorescent temperature-sensitive material and can reflect the temperature of the reaction liquid. The fluorescent temperature-sensitive material of the first fluorescent temperature-sensitive part 13 can be at least one of Rhodamine B, fluorescein, and anthocyanin. Because the thickness of the reaction cavity 12 is very small, the temperature of the reaction liquid can be quickly and uniformly distributed. At the same time, the heat transfer between the cavity wall 11 and the reaction liquid is very fast. Therefore, the temperature of the cavity wall 11 can be considered to be the same as the temperature of the reaction liquid. The first fluorescent temperature-sensitive part 13 is disposed on the outside of the carrier 1 and on the cavity wall 11. Therefore, the heat of the cavity wall 11 can be quickly conducted to the second fluorescent temperature-sensitive part 41, thereby enabling the second fluorescent temperature-sensitive part 41 to reflect the temperature of the reaction liquid in real time and accurately.

[0150] Additionally, the first fluorescent temperature-sensitive part 13 can be disposed on the heat-conducting sheet 115, thus providing a certain degree of light-blocking effect to the cavity wall 11. When the first fluorescent temperature-sensitive part 13 is directly disposed on the outside of the cavity wall 11, the cavity wall 11 can prevent the fluorescence used for temperature detection from entering the reaction chamber 12, avoiding interference from the fluorescence used for temperature detection on the detection results of the fluorescence detection device 6, and ensuring the accuracy of PCR detection. Of course, in other optional embodiments, the first fluorescent temperature-sensitive part 13 can also be disposed on the transparent material 114.

[0151] like Figure 6bAs shown, a light-blocking portion 14 is further provided between the first fluorescence temperature-sensitive part 13 and the cavity wall 11. Thus, regardless of whether the first fluorescence temperature-sensitive part 13 is located on the heat-conducting sheet 115 or on a transparent material, the light-blocking portion 14 can completely prevent the fluorescence used for temperature detection from entering the reaction chamber 12, completely avoiding interference from the fluorescence used for temperature detection on the detection results of the fluorescence detection device 6, and ensuring the accuracy of PCR detection. The light-blocking portion 14 can be black, or it can be opaque; preferably, it is made of a metallic material.

[0152] Optionally, the light-blocking part 14 can also be a black coating applied to the outside of the cavity wall 11. The specific material of the light-blocking part 14 is not limited here, as long as the light-blocking part 14 is insoluble in the coolant, can be coated on the outside of the cavity wall 11, and has a thermally conductive function. The first fluorescent temperature-sensitive part 13 is a fluorescent temperature-sensitive material layer coated on the light-blocking part 14.

[0153] like Figures 6b-7 As shown, specifically, the first fluorescent temperature-sensitive part 13 is disposed on the outer side of the first wall 111, and at least a portion of the second wall 112 is made of transparent material 114. The excitation light emitted by the fluorescence detection device 6 passes through the transparent material 114 located on the second wall 112 for detection. The first temperature detection mechanism 3 detects the temperature through the first fluorescent temperature-sensitive part 13 disposed on the first wall 111. The first wall 111 and the second wall 112 are disposed opposite to each other, thus further avoiding mutual interference between temperature detection and the detection of the content of the analyte. Of course, in other optional embodiments, at least a portion of the sidewall 113 may also be made of transparent material 114, and the fluorescence detection device 6 may perform detection through the transparent material 114 of the sidewall 113.

[0154] like Figure 7 As shown, optionally, the first temperature detection mechanism 3 includes a first fluorescent transceiver component 33 and a first controller. The first fluorescent transceiver component 33 is used to emit excitation light of a preset intensity to the first fluorescent temperature-sensitive part 13 and to receive and detect the intensity of the excitation light at the first fluorescent temperature-sensitive part 13. The first controller is electrically connected to the first fluorescent transceiver component 33 and is used to convert the light intensity signal detected by the first fluorescent transceiver component 33 into a temperature signal.

[0155] It is understandable that the temperature measurement principle of the first temperature detection mechanism 3 and the second temperature detection mechanism 4 is the same. The difference between the two is that the first temperature detection mechanism 3 does not need to be inserted into the reaction solution to detect the temperature; the first temperature detection mechanism 3 can detect the temperature from one side of the first fluorescent temperature-sensitive part 13. In this embodiment, the first fluorescent temperature-sensitive part 13 can achieve accurate temperature measurement with high sensitivity.

[0156] Specifically, the first fluorescence transmitting and receiving assembly 33 may include a first light source, a first incident channel 31, a first receiving channel 32, and a first detector 34. The first light source emits excitation light of a preset intensity. The first incident channel 31 is connected to the first light source and transmits the excitation light from the first light source to the first fluorescence temperature-sensitive part 13. The first receiving channel 32 transmits the excitation light from the first fluorescence temperature-sensitive part 13 to the first detector 34. The first detector 34 can detect the intensity of the excitation light transmitted by the first receiving channel 32.

[0157] like Figure 7 As shown, optionally, the first incident channel 31 and the first receiving channel 32 can be optical fibers. Optionally, the first exit end of the first incident channel 31 and the first incident end of the first receiving channel 32 are both located on one side of the first fluorescence temperature-sensitive part 13 to ensure the accuracy of the detection results. Furthermore, the first incident channel 31 and the first receiving channel 32 can be arranged adjacent to each other to further improve detection accuracy. Figure 8 As shown, the first incident channel 31 and the first receiving channel 32 may not be physical objects, but rather channels for the propagation of light in space.

[0158] Example 4

[0159] This embodiment provides a PCR amplification method. The PCR amplification method of this embodiment four can be performed using the vector 1 provided in embodiment one and the PCR detection system provided in embodiment two, or the PCR amplification method of this embodiment four can be performed using the vector 1 provided in embodiment one and the PCR detection system provided in embodiment three.

[0160] PCR amplification methods include:

[0161] The coolant flows continuously within the cooling mechanism to continuously cool carrier 1;

[0162] The electromagnetic heating mechanism generates a magnetic field, which heats the carrier 1.

[0163] The electromagnetic heating mechanism generates a magnetic field, causing the carrier 1 to heat up rapidly, which in turn raises the temperature of the reaction solution within the carrier 1. The cooling mechanism cools the carrier 1, thereby lowering the temperature of the reaction solution within it. The electromagnetic heating and cooling mechanisms maintain the reaction solution at the denaturation, extension, and annealing temperatures required for the PCR reaction for a period of time. The coolant within the cooling mechanism flows continuously, ready to provide cooling at any time for rapid cooling.

[0164] The electromagnetic heating mechanism heats the carrier 1 via a non-contact electromagnetic field. Since the electromagnetic heating mechanism does not come into contact with the carrier 1, even if condensation occurs on the carrier 1 during heating or cooling, it will not cause a short circuit in the electromagnetic heating mechanism's circuitry, thus improving the reliability of the equipment. Furthermore, because the electromagnetic heating mechanism can heat the reaction liquid within the carrier 1 without direct contact, the time required for heat conduction between the carrier 1 and the electromagnetic heating mechanism is saved, resulting in rapid heating and accelerating the PCR reaction process and efficiency. This solves the problems of long detection times and slow heating / cooling rates caused by the large heat capacity of the heater and the high thermal resistance between the heater and the carrier 1.

[0165] Optionally, the coolant continuously flows within the cooling mechanism to continuously cool the carrier 1, including:

[0166] Carrier 1 is in direct contact with the coolant.

[0167] The carrier 1 is in direct contact with the coolant, eliminating the need for other heat-conducting structures between the carrier 1 and the coolant, thus enabling the carrier 1 to cool down rapidly.

[0168] Optionally, the temperature of carrier 1 can be adjusted by regulating the power of the electromagnetic heating mechanism and / or by regulating the temperature and / or flow rate of the coolant in the cooling mechanism. For example, when the reaction liquid needs to be cooled, the power of the electromagnetic heating mechanism can be reduced, such as by increasing the duty cycle or decreasing the current, and / or increasing the flow rate of the coolant; when the temperature needs to be increased, the power of the electromagnetic heating mechanism can be increased, such as by decreasing the duty cycle or increasing the current, and / or decreasing the flow rate of the coolant.

[0169] The PCR detection method provided in this embodiment can achieve rapid heating and cooling. For example, compared to traditional contact heaters, where the cooling mechanism needs to cool the heater first, and then the heater cools the carrier 1, resulting in a long heat conduction time, the cooling mechanism in this embodiment does not need to cool the electromagnetic heating mechanism but directly cools the carrier 1, thus accelerating the cooling speed. Furthermore, in a contact heater, heat conduction between the carrier 1 and the heater takes time, and the heater itself also needs time to heat up. In this embodiment, the electromagnetic heating mechanism directly generates eddies within the carrier 1, which heats the carrier 1 and subsequently the reaction solution within it, thus significantly reducing the time required for heat conduction and improving the heating efficiency of the reaction solution. Moreover, the continuous flow of coolant eliminates the need to introduce coolants of different temperatures during the PCR reaction, greatly reducing the reaction time of the cooling mechanism and facilitating rapid heating and cooling.

[0170] PCR amplification methods also include:

[0171] The temperature of the reaction solution is detected by using a fluorescent thermosensitive material and the intensity of excitation light. This temperature detection method is highly sensitive and can achieve accurate temperature measurement.

[0172] The temperature of the reaction solution is detected using fluorescent thermosensitive materials and the intensity of excitation light, including:

[0173] Continuous emission of excitation light into a fluorescent thermosensitive material;

[0174] Collect the excitation light from the fluorescent temperature-sensitive material.

[0175] like Figure 5 As shown, the intensity of the collected excitation light is obtained, and the temperature of the reaction solution is obtained from the intensity of the collected excitation light.

[0176] By continuously emitting excitation light to the fluorescent thermosensitive material, the temperature of carrier 1 can be detected in real time.

[0177] A temperature detection mechanism emits monochromatic excitation light with wavelength λ0. When this excitation light illuminates a fluorescent thermosensitive material, the fluorescent molecules with wavelength λ0 generate light of a different wavelength as the temperature of the material changes. This results in a decrease in the intensity of the excitation light with wavelength λ0. The intensity I of the weakened excitation light is collected, and the relationship between intensity I and temperature T is as follows: Figure 5 As shown, and satisfies the formula: The units for T1 and T2 are K.

[0178] PCR detection methods are not affected by the monochromaticity of the excitation light (i.e., the wavelength distribution of monochromatic light), which can improve detection accuracy and sensitivity, and ensure the accuracy of temperature control.

[0179] Although the present invention has been described in detail above with general descriptions, specific embodiments, and experiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A PCR detection system, characterized in that, It includes an electromagnetic heating mechanism and a cooling mechanism. The electromagnetic heating mechanism is used to heat the carrier, and the cooling mechanism is used to cool the carrier. The carrier includes a cavity wall (11) that surrounds a reaction cavity (12) for containing a reaction liquid. At least a portion of the cavity wall (11) can be heated by an electromagnetic field. At least a portion of the cavity wall (11) is formed by a heat-conducting plate (115) that can be heated by an electromagnetic field. The PCR detection system also includes a second temperature detection mechanism (4), which is used to detect the temperature of the reaction solution inside the carrier; The second temperature detection mechanism (4) includes: The second fluorescent temperature-sensitive part (41) includes a fluorescent temperature-sensitive material, and the second fluorescent temperature-sensitive part (41) can be located in the reaction liquid inside the reaction chamber (12) of the carrier; The second fluorescent transceiver assembly (45) is used to emit excitation light of a preset intensity to the second fluorescent temperature-sensitive part (41) and to receive and detect the intensity of the excitation light at the second fluorescent temperature-sensitive part (41). The second controller is electrically connected to the second fluorescent transducer (45) and is used to convert the light intensity signal detected by the second fluorescent transducer (45) into a temperature signal; The second fluorescent transmitter assembly (45) includes: The second incident channel (42) is used to transmit the excitation light to the second fluorescence temperature-sensitive part (41). The second receiving channel (43) is used to transmit the excitation light at the second fluorescent temperature-sensitive part (41); The second exit end (421) of the second incident channel (42), the second incident end of the second receiving channel (43) and the second fluorescent temperature-sensitive part (41) can all be located in the reaction liquid of the reaction chamber (12); The second temperature detection mechanism (4) further includes a light-blocking covering part (44), at least the second emission end (421) of the second incident channel (42), at least the second incident end of the second receiving channel (43) and the second fluorescent temperature-sensitive part (41) are all disposed in the light-blocking covering part (44).

2. The PCR detection system according to claim 1, characterized in that, The thickness of the heat-conducting sheet (115) is 10-100 μm.

3. The PCR detection system according to claim 1, characterized in that, At least part of the cavity wall (11) is made of a transparent material (114).

4. The PCR detection system according to claim 3, characterized in that, The transparent material (114) is polydimethylsiloxane, polypropylene, plexiglass or polycarbonate.

5. The PCR detection system according to any one of claims 1-4, characterized in that, The reaction chamber (12) and / or the carrier are flat structures.

6. The PCR detection system according to claim 5, characterized in that, The flat structure refers to the reaction chamber (12) or the carrier having a dimension perpendicular to its thickness direction that is greater than its thickness direction dimension.

7. The PCR detection system according to claim 6, characterized in that, The ratio of the dimension of the reaction chamber (12) or the carrier perpendicular to its thickness direction to its dimension in the thickness direction is greater than 5:

1.

8. The PCR detection system according to claim 7, characterized in that, The size ratio is 50:1 to 100:

1.

9. The PCR detection system according to claim 1, characterized in that, The second fluorescent temperature-sensitive part (41) includes a fluorescent temperature-sensitive material coated on the second exit end (421) of the second incident channel (42); and / or, the second fluorescent temperature-sensitive part (41) is connected to the second exit end (421) of the second incident channel (42) and extends from the second incident channel (42).

10. The PCR detection system according to claim 1 or 9, characterized in that, The cooling mechanism includes a container (5) in which coolant is continuously circulated, and the carrier can be inserted into the coolant.

11. The PCR detection system according to claim 10, characterized in that, The electromagnetic heating mechanism includes a coil (2) for carrying current.

12. The PCR detection system according to claim 11, characterized in that, The coil (2) is disposed on the upper or lower side of the container (5), or the coil (2) is sleeved on the outside of the container (5), or the coil (2) is disposed inside the container (5), and / or The PCR detection system also includes a fluorescence detection device (6), which is used to detect the content of the analyte in the reaction solution.

13. The PCR detection system according to claim 12, characterized in that, The fluorescence detection device (6) is located outside the container (5).

14. The PCR detection system according to claim 12, characterized in that, The fluorescence detection device (6) is located on one side of the axial direction or the radial direction of the coil (2).

15. A PCR amplification method, characterized in that, The PCR amplification method is performed using the PCR detection system according to any one of claims 1-14, the PCR amplification method comprising: The coolant flows continuously within the cooling system to continuously cool the medium. The electromagnetic heating mechanism generates a magnetic field, which heats the carrier.

16. The PCR amplification method according to claim 15, characterized in that, By adjusting the power of the electromagnetic heating mechanism, and / or The temperature of the carrier is adjusted by regulating the temperature and / or flow rate of the coolant in the cooling mechanism; and / or The coolant flows continuously within the cooling mechanism to continuously serve as a carrier. (1) Refrigeration includes: The carrier is in direct contact with the coolant.

17. The PCR amplification method according to claim 15 or 16, characterized in that, The PCR amplification method further includes: The temperature of the reaction solution is detected by using a fluorescent thermosensitive material and the intensity of excitation light.

18. The PCR amplification method according to claim 17, characterized in that, The method of detecting the temperature of the reaction solution by means of a fluorescent thermosensitive material and the intensity of excitation light includes: The fluorescent thermosensitive material continuously emits excitation light; Collect the excitation light from the fluorescent temperature-sensitive material. The intensity of the collected excitation light is obtained, and the temperature of the reaction solution is obtained from the intensity of the collected excitation light.

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