Vector, PCR (Polymerase Chain Reaction) detection system and PCR amplification method

Abstract

The invention relates to a carrier, a PCR (Polymerase Chain Reaction) detection system and a PCR amplification method, the carrier comprises a cavity wall, the cavity wall defines a reaction cavity for accommodating reaction liquid, and at least part of the cavity wall can be heated by an electromagnetic field. The PCR detection system comprises an electromagnetic heating mechanism and a cooling mechanism, the electromagnetic heating mechanism is used for heating the carrier, and the cooling mechanism is used for cooling the carrier. The PCR amplification method is executed by using the carrier and the PCR detection system, and the PCR amplification method comprises the following steps: cooling liquid continuously flows in the cooling mechanism so as to continuously refrigerate the carrier; the electromagnetic heating mechanism generates a magnetic field and heats the carrier through the magnetic field.

Description

Technical Field

[0001] The present 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 Art

[0002] PCR (polymerase chain reaction) refers to a molecular biology experimental method for enzymatically synthesizing specific DNA fragments in vitro, which mainly consists of repeated thermal cycles of three steps: high-temperature denaturation, low-temperature annealing, and thermophilic extension. Before the PCR reaction, the reaction solution needs to be placed in a carrier, where the reaction solution is composed of samples such as collected throat swabs or nasal swabs and reagents for PCR reactions. During the PCR reaction, the reaction solution needs to be heated or cooled by the temperature module of the PCR instrument, so that the reaction solution circulates in the high-temperature denaturation, low-temperature annealing, and thermophilic extension stages.

[0003] The existing technology has a long detection time and a slow temperature rise and fall speed, which is mainly caused by factors such as the large heat capacity of the heater, the large thermal resistance between the heater and the carrier, and the slow thermal balance of the carrier.

[0004] The existing technology uses contact heating, which easily generates condensed water during the heating and cooling process. The condensed water can cause circuit short circuits and reduce the reliability of the equipment.

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

[0006] Prior art often uses fluorescence detection to determine the content of an analyte. The prior art also discloses methods for determining the temperature of solid objects using fluorescence detection. However, when this temperature detection method is applied to measuring component content, interference between the fluorescence used to detect temperature and the fluorescence used to detect component content is unavoidable, resulting in poor detection accuracy. Furthermore, prior art methods for detecting temperature through fluorescence have low sensitivity. Summary of the Invention

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

[0008] To achieve the above objectives, the present invention provides a carrier in a first aspect, comprising a cavity wall, wherein the cavity wall forms a reaction cavity for accommodating a reaction liquid, and at least a portion of the cavity wall can be heated by an electromagnetic field.

[0009] Optionally, the carrier further includes a temperature sensing portion, and the temperature sensing portion is used to reflect the temperature of the reaction liquid.

[0010] Optionally, the temperature sensing portion includes a first fluorescent temperature-sensitive portion arranged on the outside of the cavity wall, the first fluorescent temperature-sensitive portion includes a fluorescent temperature-sensitive material, and the first fluorescent temperature-sensitive portion can reflect the temperature of the reaction liquid.

[0011] Optionally, a light-isolating portion is further provided between the first fluorescent temperature-sensitive portion and the cavity wall.

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

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

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

[0015] Optionally, the thickness of the thermal conductive sheet is 10-100 μm.

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

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

[0018] Optionally, the reaction chamber and / or the carrier are flat structures.

[0019] Optionally, the flat structure means that the dimension of the reaction chamber or the carrier perpendicular to its thickness direction is larger than its thickness direction.

[0020] Optionally, the ratio of the dimension of the reaction chamber or the carrier perpendicular to its thickness direction to its 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 solve at least one of the above technical problems.

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

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

[0026] Optionally, the PCR detection system further includes a temperature detection mechanism, which is used to detect the temperature of the reaction solution in the carrier.

[0027] Optionally, the temperature detection mechanism is a first temperature detection mechanism, which detects the temperature of the reaction liquid by detecting the fluorescence intensity at a temperature sensing portion of the carrier.

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

[0029] a first fluorescent light receiving and emitting component, configured to emit excitation light of a preset intensity toward the temperature sensing portion and to receive and detect the intensity of the excitation light at the temperature sensing portion;

[0030] The first controller is electrically connected to the first fluorescent light receiving and emitting component, and is used to convert the light intensity signal detected by the first fluorescent light receiving and emitting component into a temperature signal.

[0031] Optionally, the first fluorescent light receiving assembly includes:

[0032] a first light source, configured to emit the excitation light of a preset intensity;

[0033] a first incident channel connected to the first light source and configured to transmit the excitation light of the first light source to the temperature sensing portion;

[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 portion to the first detector, and the first detector can detect the intensity of the excitation light transmitted by the first receiving channel.

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

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

[0037] a second fluorescent temperature-sensitive portion, comprising a fluorescent temperature-sensitive material, wherein the second fluorescent temperature-sensitive portion can be located in the reaction liquid in the reaction chamber of the carrier;

[0038] a second fluorescent light-receiving and transmitting component configured to transmit excitation light of a preset intensity to the second fluorescent temperature-sensitive portion and to receive and detect the intensity of the excitation light at the second fluorescent temperature-sensitive portion;

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

[0040] Optionally, the second fluorescent light receiving assembly includes:

[0041] a second incident channel, for transmitting the excitation light to the second fluorescent temperature-sensitive portion;

[0042] a second receiving channel, the second receiving channel being used to transmit the excitation light at the second fluorescent temperature-sensitive portion;

[0043] The second output end of the second incident channel, the second incident end of the second receiving channel, and the second fluorescent temperature-sensitive portion can all be located in the reaction liquid of the reaction chamber.

[0044] Optionally, the second temperature detection mechanism further includes a light-isolating covering portion, and 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 portion are all arranged in the light-isolating covering portion.

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

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

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

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

[0049] The PCR detection system further comprises 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 arranged outside the container.

[0051] Optionally, the fluorescence detection device is located on one axial side or one radial side 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 technical problems.

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

[0054] A PCR amplification method is performed using the above-mentioned vector and the above-mentioned PCR detection system, and the PCR amplification method includes:

[0055] The coolant flows continuously in the cooling mechanism to continuously cool the carrier;

[0056] The electromagnetic heating mechanism generates a magnetic field and heats the carrier through the magnetic field.

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

[0058] Adjusting the temperature of the carrier by adjusting the temperature and / or flow rate of the cooling liquid in the cooling mechanism; and / or

[0059] The coolant continuously flows in 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 comprises:

[0062] The temperature of the reaction solution is detected by the fluorescent temperature-sensitive material and the intensity of the excitation light.

[0063] Optionally, detecting the temperature of the reaction solution by using a fluorescent temperature-sensitive material and the intensity of the excitation light includes:

[0064] continuously emitting excitation light toward the fluorescent temperature-sensitive material;

[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 liquid is obtained according to the intensity of the collected excitation light.

[0067] As can be seen from the above, the technical solution provided by the present invention is that the cavity wall can be heated by an electromagnetic field. Therefore, the carrier can be heated by a non-contact electromagnetic field. Since the electromagnetic heating mechanism that generates the electromagnetic field does not contact the carrier, even if the carrier produces condensed water during the heating and cooling process, the circuit of the electromagnetic heating mechanism will not be short-circuited, thereby improving the reliability of the equipment. The electromagnetic heating mechanism that generates the electromagnetic field can heat the reaction liquid in the carrier without contact with the carrier. Therefore, when heating the carrier, the time required for heat conduction between the carrier and the electromagnetic heating mechanism is saved, the heating efficiency is improved, and the PCR reaction process and PCR reaction efficiency are accelerated. It solves the problems of long detection time and slow heating and cooling speed caused by the large heat capacity of traditional heaters and the large thermal resistance between the heater and the carrier.

[0068] The carrier is directly formed by the heat conducting sheet or the heat conducting sheet and the transparent material, which reduces the heat capacity of the carrier, increases the temperature of the reaction liquid quickly, reduces the carrier cost, and simplifies the process.

[0069] The carrier may further include a first fluorescent temperature-sensitive portion, and the first temperature detection mechanism detects the temperature of the reaction liquid by detecting the fluorescence intensity at the first fluorescent temperature-sensitive portion of the carrier. Since the thickness of the reaction chamber is very small, the temperature of the reaction liquid can be quickly and evenly distributed throughout the reaction liquid. At the same time, the heat transfer between the cavity wall and the reaction liquid is very fast, so the temperature of the cavity wall can be considered to be consistent with the temperature of the reaction liquid. The first fluorescent temperature-sensitive portion is arranged on the outside of the carrier and on the cavity wall. Therefore, the heat of the cavity wall can be quickly transferred to the second fluorescent temperature-sensitive portion, thereby allowing the second fluorescent temperature-sensitive portion to accurately reflect the temperature of the reaction liquid in real time.

[0070] The cavity wall has a certain light-isolating effect. When the first fluorescent temperature-sensitive part is directly arranged on the outside of the cavity wall, the cavity wall can prevent the fluorescence used for temperature detection from entering the reaction chamber, thereby avoiding the fluorescence used for temperature detection from interfering with the detection results of the fluorescence detection device and ensuring the accuracy of PCR detection.

[0071] A light-isolating portion is also provided between the first fluorescent temperature-sensitive portion and the cavity wall. When light leaks from the cavity wall, the light-isolating portion completely blocks the fluorescent light used for temperature detection from entering the reaction chamber, completely preventing it from interfering with the detection results of the fluorescence detection device and ensuring PCR detection accuracy.

[0072] The reaction chamber and / or carrier has a flat structure, which makes the reaction liquid in 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 a consistent level in a very short time, thereby improving the thermal equilibrium speed of the carrier, and greatly improving the reaction liquid heating and cooling speed and detection efficiency.

[0073] The PCR detection system includes an electromagnetic heating mechanism and a cooling mechanism. The electromagnetic heating mechanism heats the carrier, while the cooling mechanism cools the carrier. The electromagnetic heating mechanism generates an alternating magnetic field. Through eddy current heating, eddy currents are induced in the thermal conductive sheet, thereby increasing the temperature of the carrier and, in turn, the reaction solution within the carrier. The electromagnetic heating mechanism heats the carrier using a non-contact electromagnetic field. Therefore, even if condensation forms during the heating and cooling process, it does not short-circuit the heating mechanism's circuit, thereby improving device reliability. Because the electromagnetic heating mechanism heats the reaction solution within the carrier without contact between the heating mechanism and the carrier, the non-contact heating mechanism reduces the time required for heat transfer between the carrier and conventional heaters, as well as the time required for the heaters themselves to heat up. This allows the carrier and reaction solution to heat up quickly, improving heating efficiency and accelerating the PCR reaction process and efficiency. This solves the problem of long detection times and slow heating and cooling speeds 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 reducing the temperature of the reaction liquid in the carrier. The electromagnetic heating mechanism and the cooling mechanism keep the reaction liquid at the denaturation temperature, extension temperature and annealing temperature during the PCR reaction and maintain them for a period of time.

[0075] 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 reducing the current, and / or increasing the coolant flow rate. When the power of the electromagnetic heating mechanism changes, the eddy currents within the carrier change instantaneously, and the carrier's own heat generation also changes accordingly. Therefore, the PCR detection system can more easily and quickly adjust the temperature of the carrier and the reaction liquid. The cooling mechanism includes a container in which coolant continuously flows. The carrier can be inserted into the coolant, that is, the coolant directly contacts the carrier. During the PCR reaction, the coolant continuously flows, so the cooling mechanism is always ready to cool the carrier, allowing the carrier to be cooled quickly when cooling is required.

[0076] The first fluorescent receiving component (or the second fluorescent receiving component (i.e., the second light source)) emits monochromatic excitation light with a wavelength of λ0. After the excitation light emitted by the first fluorescent receiving component (or the second fluorescent receiving component) is irradiated by the first fluorescent temperature-sensitive portion (or the second fluorescent temperature-sensitive portion), the fluorescent molecules with a wavelength of λ0 will produce light of another wavelength as the temperature of the first fluorescent temperature-sensitive portion (or the second fluorescent temperature-sensitive portion) changes, thereby causing the intensity of the excitation light with a wavelength of λ0 to weaken. 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 not affected by the monochromaticity (i.e., the wavelength distribution of the monochromatic light) of the excitation light emitted by the first fluorescent receiving component (or the second fluorescent receiving component (i.e., the second light source)), and can improve detection accuracy and sensitivity, ensuring the accuracy of temperature control.

[0077] The second output end of the second incident channel, the second input end of the second receiving channel, and the second fluorescent temperature-sensitive portion can all be located in the reaction liquid of the reaction chamber. Among them, the second fluorescent temperature-sensitive portion is located in the reaction liquid of the reaction chamber, which can ensure that the temperature of the second fluorescent temperature-sensitive portion is consistent with that of the reaction liquid, thereby improving the temperature detection accuracy and thus improving the PCR detection accuracy; the second output end of the second incident channel and the second incident end of the second receiving channel are located in the reaction liquid of the reaction chamber, which can ensure that the distance between them and the second fluorescent temperature-sensitive portion is very small, thereby improving the accuracy of fluorescence intensity detection and thus ensuring the temperature detection accuracy.

[0078] The second temperature detection mechanism also includes a light-isolating cladding portion, within which at least the second output end of the second incident channel, at least the second incident end of the second receiving channel, and the second fluorescent temperature-sensitive portion are disposed. In this embodiment, the light used for temperature detection is distributed within the light-isolating cladding portion and does not leak outside the light-isolating cladding portion. This prevents light from the temperature detection mechanism from interfering with the detection results of the fluorescence detection device, thereby ensuring PCR detection accuracy.

[0079] PCR amplification methods include:

[0080] The coolant flows continuously in the cooling mechanism to continuously cool the carrier;

[0081] The electromagnetic heating mechanism generates a magnetic field and heats the carrier through the magnetic field.

[0082] The electromagnetic heating mechanism generates a magnetic field, causing the carrier to heat up rapidly, thereby increasing the temperature of the reaction solution within the carrier. The cooling mechanism cools the carrier, thereby reducing the temperature of the reaction solution within the carrier. The electromagnetic heating and cooling mechanisms maintain the reaction solution at the denaturation, extension, and annealing temperatures, respectively, during the PCR reaction, for a period of time. The coolant in the cooling mechanism continuously flows, allowing for ready cooling at any time, achieving rapid cooling.

[0083] The electromagnetic heating mechanism heats the carrier via a non-contact electromagnetic field. Since the electromagnetic heating mechanism does not come into contact with the carrier, even if condensation forms during the heating and cooling process, this will not short-circuit the electromagnetic heating mechanism's circuit, thereby improving device reliability. Furthermore, because the electromagnetic heating mechanism heats the reaction solution within the carrier without contact, the time required for heat conduction between the carrier and the electromagnetic heating mechanism is reduced during heating, enabling rapid temperature rise and accelerating the PCR reaction process and efficiency. This solves the problem of long detection times and slow heating and cooling speeds caused by the large thermal capacity of the heater and the high thermal resistance between the heater and the carrier. BRIEF DESCRIPTION OF THE DRAWINGS

[0084] Figure 1 Schematic diagram of the structure of the carrier provided by an embodiment of the present invention;

[0085] Figure 2a Schematic diagram of the structure of a PCR detection system provided by an embodiment of the present invention;

[0086] Figure 2b is a temperature rise curve of the reaction solution when the reaction solution in the carrier is heated using the PCR detection system provided in this embodiment;

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

[0088] Figure 4 This is a schematic structural diagram of the connection between the second emission end and the second fluorescent temperature-sensitive portion provided by an embodiment of the present invention;

[0089] Figure 5 is a corresponding curve between the collected excitation light intensity (I) and temperature (T) provided in an embodiment of the present invention;

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

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

[0092] Figure 7 Schematic diagram of another PCR detection system provided by an embodiment of the present invention;

[0093] Figure 8 It is a structural schematic diagram of another PCR detection system provided by 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. Heat conducting sheet; 12. Reaction chamber; 13. First fluorescent temperature-sensitive portion; 14. Light-isolating portion;

[0096] 2. Coil;

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

[0098] 4. Second temperature detection mechanism; 41. Second fluorescent temperature-sensitive portion; 42. Second incident channel; 421. Second emission end; 43. Second receiving channel; 44. Light-isolating covering portion; 45. Second fluorescent light-receiving assembly; 46. Second detector;

[0099] 5. Container;

[0100] 6. Fluorescence detection device. DETAILED DESCRIPTION

[0101] The technical solution of the present invention will be further described below with reference to the accompanying drawings and through specific embodiments. It will be understood that the specific embodiments described herein are intended only to illustrate the present invention and are not intended to limit the present invention. It should also be noted that, for ease of description, the accompanying drawings only show portions relevant to the present invention, not all of them.

[0102] Some directional words are defined in the present invention. Unless otherwise specified, the directional words used, such as "up", "down", "left", "right", "inside" and "outside", are used for ease of understanding and therefore do not constitute a limitation on the scope of protection of the present invention.

[0103] In the present invention, unless otherwise expressly specified or limited, a first feature being "above" or "below" a second feature may include the first and second features being in direct contact, or may include the first and second features being in contact not directly but through another feature between them. Furthermore, a first feature being "above," "above," and "above" a second feature may include the first feature being directly above or obliquely above the second feature, or may simply mean that the first feature is higher in level than the second feature. A first feature being "below," "below," and "below" a second feature may include the first feature being directly below or obliquely below the second feature, or may simply mean that the first feature is lower in level than the second feature.

[0104] In the description of the present invention, unless otherwise expressly specified or limited, the terms "connected," "connected," and "fixed" should be understood in a broad sense. For example, they may refer to fixed connections, detachable connections, or integration; mechanical connections or electrical connections; direct connections or indirect connections through an intermediate medium; and internal communication between two components or interaction between two components. Those skilled in the art will understand the specific meanings of the above terms in the present invention in specific circumstances.

[0105] Example 1

[0106] This embodiment provides a vector 1 for use in PCR reaction 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 accommodating a 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 the carrier 1 generates condensed water 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 in the carrier 1 without contact between 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, the heating efficiency is improved, and the PCR reaction process and PCR reaction efficiency are accelerated. The problem of long detection time and slow heating and cooling speed caused by the large heat capacity of the traditional heater and the large thermal resistance between the heater and the carrier 1 is solved.

[0109] Optionally, at least part 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 thickness of the heat conducting sheet 115 is small and has a heat conducting function. Therefore, when the reaction liquid needs to be cooled, the heat of the reaction liquid can also be quickly transferred to the outside of the heat conducting sheet 115. The heat conducting sheet 115 can also be called a heat conducting 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 heat conduction efficiency of the carrier 1, improve the detection efficiency, and shorten the detection time.

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

[0111] Optionally, the chamber wall 11 includes a first wall 111 and a second wall 112 disposed opposite 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 enclose the reaction chamber 12. It is understood that the side wall 113 is annular, which means that the side wall 113 extends continuously until both ends are closed. The annular shape can be a polygonal ring such as a circular ring, an elliptical ring, a square ring, etc., to enclose the space between the first wall 111 and the second wall 112.

[0112] The reaction chamber 12 and / or the carrier 1 are flat structures, which makes the thickness of the reaction liquid in the reaction chamber 12 very thin, such as the thickness of the reaction liquid can be 0.3-1 mm. 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 a uniform level in a very short time, thereby improving the thermal equilibrium speed of the carrier 1, and greatly improving the heating and cooling speed of the reaction liquid and the detection efficiency.

[0113] Optionally, when the carrier 1 has a flat structure, it can accommodate a flat reaction chamber 12. Specifically, a flat structure means that the dimension of the reaction chamber 12 or the carrier 1 in the thickness direction is smaller than the dimension perpendicular to the thickness direction. As an 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 of the carrier 1 or the reaction chamber 12 in the thickness direction is much smaller than the dimension perpendicular to the thickness direction, such as a dimension ratio of 50:1 to 100:1. As an example, the dimension ratio is 90:1. As an example, the reaction chamber 12 is a rectangular parallelepiped, and the ratio of the length to the thickness of the rectangular parallelepiped can be greater than 5:1, such as 90:1. For example, the dimension of the reaction chamber 12 in the thickness direction 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 may 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 may be polygonal, circular, or elliptical.

[0114] Optionally, at least part of the cavity wall 11 is made of a transparent material 114. For example, part of the side wall 113 (e.g. 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 The cavity wall 11 is made of a transparent material 114. Optionally, all portions of the cavity wall 11 except the transparent material 114 are made of a heat conducting sheet 115. The excitation light reacts with the fluorescent probe / dye in the reaction solution to generate fluorescence. The fluorescence can pass through the transparent material 114 and is detected by the fluorescence detection device 6 to detect the content of the analyte in the reaction solution.

[0115] Optionally, the transparent material 114 may be polydimethylsiloxane, polypropylene, organic glass or polycarbonate. PMMA, PDMS, PP and PC are optically transparent materials with good biocompatibility, and thus can meet the requirements of fluorescence detection and have no effect on the reaction sample.

[0116] The carrier 1 provided in this embodiment is directly formed by a heat conducting sheet or a heat conducting sheet and a transparent material 114, which reduces the heat capacity of the carrier 1, the reaction liquid heats up quickly, the carrier 1 has low cost, and the process is simple. For example, a flat carrier 1 can be prepared in batches using a metal sheet (metal film) material, which is low in cost and simple in process. The use of an ultra-thin carrier 1 (i.e., the dimension of the carrier 1 in the thickness direction is much smaller than the dimension in other directions) reduces the total heat capacity of the carrier 1, increases the thermal equilibrium speed of the reaction liquid and the carrier 1, and thus increases the heating and cooling speed.

[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. Through eddy current heating, eddy currents are induced in the heat conducting plate 115, thereby increasing the temperature of the carrier 1 and, in turn, rapidly increasing the temperature of the reaction liquid within the carrier 1. The electromagnetic heating mechanism heats the carrier 1 using a non-contact electromagnetic field. The electromagnetic heating mechanism does not contact the carrier 1. Therefore, even if condensed water is generated by the carrier 1 during the heating and cooling process, the electromagnetic heating mechanism circuit will not short-circuit, thereby improving the reliability of the device. Because the electromagnetic heating mechanism and the carrier 1 can heat the reaction liquid within the carrier 1 without contact, the non-contact electromagnetic heating mechanism heats the carrier 1, saving the time required for heat conduction between the carrier 1 and a conventional heater, as well as the time required for the heater itself to heat up. The carrier 1 and the reaction liquid heat up quickly, improving heating efficiency and accelerating the PCR reaction process and efficiency. This solves the problem of long detection time and slow heating and cooling speeds caused by the large heat capacity of the heater and the high thermal resistance between the heater and the carrier 1. Figure 2b As shown, after experimental verification, when the PCR detection system provided in this embodiment is used to heat the reaction liquid in the carrier 1, the heating rate of the reaction liquid can reach 100°C / s.

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

[0119] 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 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 adjust the temperature of the carrier 1 and the reaction liquid more easily and quickly.

[0120] Optionally, the cooling mechanism includes a container 5, in which a coolant continuously circulates, and the carrier 1 can be inserted into the coolant, that is, the coolant is in direct contact with the carrier 1, eliminating other heat-conducting structures between the carrier and the coolant, and can achieve rapid cooling of the carrier. Preferably, the entire carrier 1 is inserted into the coolant, and of course, a portion of the carrier 1 can also be inserted into the coolant. By inserting the carrier 1 into the coolant, the coolant continues to flow during the PCR reaction, so that the cooling mechanism is in a state that can cool the carrier 1 at any time, so that when the carrier 1 needs to be cooled, the carrier 1 can be quickly cooled.

[0121] Optionally, the electromagnetic heating mechanism includes a coil 2 for circulating current. Alternating current is passed through the coil 2 to generate a magnetic field, which in turn 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 produce, and has low cost.

[0122] Exemplarily, the coil 2 is arranged on the lower side of the container 5 (eg Figure 2a As shown), the coil 2 is arranged on the upper side of the container 5, or the coil 2 is sleeved on the outside of the container 5, and the coil 2 is located outside the container 5, thereby preventing the coil 2 from short-circuiting.

[0123] In other optional embodiments, the coil 2 may also be disposed in the container 5. Optionally, when the coil 2 is disposed in the container 5, a separate space may be provided in the container 5, which is used to accommodate the coil 2 and can isolate the coolant to avoid circuit short circuit.

[0124] The PCR detection system of this embodiment adopts a direct contact cooling method of the coolant, that is, the coolant directly contacts the carrier 1 to cool the carrier 1, and the cooling speed is fast, which shortens the PCR reaction time. The alternating magnetic field generated by the coil 2 causes eddy currents to be induced on the heat conducting plate 115, and then the carrier 1 generates heat by itself, realizing non-contact eddy current heating. The process of heat conduction between the traditional heater and the carrier 1 is not required, and the temperature rises quickly, shortening the PCR reaction time. Non-contact heating has a fast heating speed and is free of condensed water that occurs during the traditional contact heating and cooling process, thereby improving the reliability of the system.

[0125] like Figure 2a As shown, optionally, the PCR detection system further 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 is not described in detail here.

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

[0127] like Figure 2a As shown, illustratively, the fluorescence detection device 6 is located on one side of the axial direction of the coil 2, so that the PCR detection system structure is compact. For example, the fluorescence detection device 6 is arranged on the upper side of the container 5, and at least a portion of the side wall 113 of the carrier 1 located at the upper end is made of a transparent material 114. In an optional embodiment, the side wall 113 includes a top wall and a bottom wall arranged opposite to 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 can be arranged on one side of the radial direction of the coil 2, so as to reduce the interference of the magnetic field on the fluorescence detection device 6.

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

[0129] Example 2

[0130] like Figure 3 As shown, the carrier 1 and PCR detection system of the second embodiment are basically the same as those of the above-mentioned first embodiment. The difference between the two is that the temperature detection mechanism is a second temperature detection mechanism 4, which is used to detect the temperature of the reaction liquid in the carrier 1 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 portion 41, a second fluorescent light-receiving and transmitting assembly 45, and a second controller. The second fluorescent temperature-sensitive portion 41 includes 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 light-receiving and transmitting assembly 45 is configured to emit excitation light of a preset intensity I0 toward the second fluorescent temperature-sensitive portion 41 and to receive and detect the intensity of the excitation light I1 at the second fluorescent temperature-sensitive portion 41.

[0132] The second controller is electrically connected to the second fluorescence receiving and transmitting assembly 45 and is configured to convert the light intensity signal detected by the second fluorescence receiving and transmitting assembly 45 into a temperature signal. The PCR detection system may further include a control mechanism that detects the real-time temperature of the reaction solution via the second temperature detection mechanism 4 and provides feedback to the control mechanism. The control mechanism identifies the real-time temperature feedback and, based on the temperature, adjusts the temperature of the carrier 1 and the reaction solution by controlling the power of the electromagnetic heating mechanism, etc., to maintain the temperature at the denaturation temperature, extension temperature, and annealing temperature, respectively, during the PCR reaction, and maintains the temperature for a period of time, thereby achieving PCR amplification. The fluorescence detection device 6 performs fluorescence detection on the reaction solution after amplification.

[0133] Optionally, the fluorescent temperature-sensitive material can be at least one of rhodamine B, fluorescein, and cyanine. Of course, the fluorescent temperature-sensitive material can also be other materials. In this embodiment, the fluorescent temperature-sensitive material is used to detect the temperature of the reaction solution, which can achieve accurate temperature measurement with high sensitivity.

[0134] like Figure 3 As shown, optionally, the second fluorescence receiving and emitting assembly 45 includes a second incident channel 42 and a second receiving channel 43. The second incident channel 42 is used to transmit the excitation light to the second fluorescence temperature-sensitive portion 41, and the second receiving channel 43 is used to transmit the excitation light at the second fluorescence temperature-sensitive portion 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 light receiving assembly 45 may further include a second light source connected to the second incident channel 42 to emit excitation light to 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 mechanism is as follows Figure 5 As shown, the second fluorescent light receiving and transmitting component 45 (i.e., the second light source) emits a monochromatic excitation light with a wavelength of λ0. After the excitation light emitted by the second fluorescent light receiving and transmitting component 45 is irradiated on the second fluorescent temperature-sensitive portion 41, the fluorescent molecules with a wavelength of λ0 will generate light of another wavelength as the temperature of the second fluorescent temperature-sensitive portion 41 changes, thereby causing the intensity of the excitation light with a wavelength of λ0 to weaken. The second receiving channel 43 collects the intensity I of the excitation light after the intensity is weakened. The corresponding relationship between the intensity I and the temperature T is shown in FIG. Figure 5 As shown, and satisfy the formula: The unit of T1 and T2 is K.

[0137] The detection method provided in this embodiment is not affected by the monochromaticity (ie, wavelength distribution of monochromatic light) of the excitation light emitted by the second fluorescent receiving assembly 45 (ie, the second light source), and can improve detection accuracy and sensitivity, thereby ensuring the accuracy of temperature control.

[0138] The second fluorescent temperature-sensitive portion 41 includes a fluorescent temperature-sensitive material coated on the second output end 421 of the second incident channel 42; and / or Figure 4 As shown, optionally, the second fluorescent temperature-sensitive portion 41 is connected to the second fluorescent light-receiving assembly 45. Specifically, the second fluorescent temperature-sensitive portion 41 is connected to the second output end 421 of the second incident channel 42 and extends from the second incident channel 42. The arrangement of the second fluorescent temperature-sensitive portion 41 can ensure that the excitation light incident on the second incident channel 42 is irradiated on the second fluorescent temperature-sensitive portion 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, thereby improving detection accuracy.

[0139] The second output end 421 of the second incident channel 42, the second incident end of the second receiving channel 43, and the second fluorescent temperature-sensitive portion 41 can all be located in the reaction liquid of the reaction chamber 12. Among them, the second fluorescent temperature-sensitive portion 41 is located in the reaction liquid of the reaction chamber 12, which can ensure that the temperature of the second fluorescent temperature-sensitive portion 41 is consistent with that of the reaction liquid, thereby improving the temperature detection accuracy and thus improving the PCR detection accuracy; the second output end 421 of the second incident channel 42 and the second incident end of the second receiving channel 43 are located in the reaction liquid of the reaction chamber 12, which can ensure that the distance between them and the second fluorescent temperature-sensitive portion 41 is very small, thereby improving the accuracy of fluorescence intensity detection and thus ensuring the temperature detection accuracy.

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

[0141] Preferably, the second temperature detection mechanism 4 further includes a light-isolating covering portion 44, wherein at least the second output 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 portion 41 are all disposed within the light-isolating covering portion 44. In this embodiment, the light used for temperature detection is uniformly distributed within the light-isolating covering portion 44 and does not leak outside the light-isolating covering portion 44. This prevents the light from the temperature detection mechanism from interfering with the detection results of the fluorescence detection device 6, thereby ensuring PCR detection accuracy.

[0142] Alternatively, the light-isolating covering 44 may be a light-proof flexible bag, for example, made of polypropylene (PP) or polycarbonate (PC). The second fluorescent temperature-sensitive portion 41 may also be a fluorescent liquid contained within the light-isolating covering 44. The second output end 421 of the second incident channel 42 and the second incident end of the second receiving channel 43 may extend into the fluorescent liquid or be located above the fluorescent liquid.

[0143] Optionally, a socket is provided on the sidewall 113 of the carrier 1, through which the portion of the second temperature detection mechanism 4 located within the reaction chamber 12 can be inserted. Specifically, the socket can provide an interference fit 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. Exemplarily, the portion of the second temperature detection mechanism 4 located within the reaction chamber 12 includes the second output end 421 of the second incident channel 42, the second incident end of the second receiving channel 43, the second fluorescent temperature-sensitive portion 41, and the light-isolating coating 44.

[0144] Optionally, the second fluorescence receiving and transmitting assembly 45 may further include a first detector 46, connected to the second receiving channel 43, for detecting the intensity of the excitation light transmitted by the second receiving channel 43. The fluorescence detection device 6 includes a fluorescence detection detector for detecting the intensity of the fluorescence. In this embodiment, the specific positions of the fluorescence detection detector and the first detector 46 are not limited; as long as the first temperature detection mechanism and the fluorescence detection device 6 are both capable of detecting the corresponding fluorescence intensity, it is sufficient.

[0145] The detector may include a silicon photomultiplier tube, a photon detector or a photomultiplier tube. The detector using the silicon photomultiplier tube, the photon detector or the photomultiplier tube has high sensitivity and can achieve ultrafast, high-sensitivity detection of fluorescence signals in milliseconds when performing fluorescence detection.

[0146] Example 3

[0147] like Figure 6a-Figure 7 As shown, the carrier 1 and PCR detection system of this third embodiment are substantially the same as those of the first embodiment described above. The difference between the two is that the carrier 1 is further provided with a structure for detecting temperature. The temperature detection mechanism is a first temperature detection mechanism 3. The first temperature detection mechanism 3 is used to detect the temperature of the reaction solution within the carrier 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 of the second embodiment.

[0148] The carrier 1 may further include a temperature sensing portion, 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 portion includes a first fluorescent temperature-sensitive portion 13, and the first temperature detection mechanism 3 detects the temperature of the reaction liquid by detecting the fluorescence intensity at the first fluorescent temperature-sensitive portion 13 of the carrier 1. Optionally, the first fluorescent temperature-sensitive portion 13 is arranged on the outside of the cavity wall 11, and the first fluorescent temperature-sensitive portion 13 includes a fluorescent temperature-sensitive material, and the first fluorescent temperature-sensitive portion 13 can reflect the temperature of the reaction liquid. The fluorescent temperature-sensitive material of the first fluorescent temperature-sensitive portion 13 can be at least one of rhodamine B, fluorescein, and cyanine. Since the thickness of the reaction chamber 12 is very small, the temperature of the reaction liquid can be quickly and evenly 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 regarded as consistent with the temperature of the reaction liquid. The first fluorescent temperature-sensitive portion 13 is arranged 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 portion 41, thereby allowing the second fluorescent temperature-sensitive portion 41 to accurately reflect the temperature of the reaction liquid in real time.

[0150] In addition, the first fluorescent temperature-sensitive portion 13 can be disposed on the heat-conducting sheet 115, so that the cavity wall 11 has a certain light-isolating effect. When the first fluorescent temperature-sensitive portion 13 is directly disposed outside the cavity wall 11, the cavity wall 11 can prevent the fluorescence used for temperature detection from entering the reaction chamber 12, thereby preventing the fluorescence used for temperature detection from interfering with the detection results of the fluorescence detection device 6, thereby ensuring the accuracy of PCR detection. Of course, in other optional embodiments, the first fluorescent temperature-sensitive portion 13 can also be disposed on the transparent material 114.

[0151] like Figure 6bAs shown, further, a light-isolating portion 14 is provided between the first fluorescent temperature-sensitive portion 13 and the cavity wall 11. Thus, regardless of whether the first fluorescent temperature-sensitive portion 13 is disposed on the thermal conductive sheet 115 or a transparent material, the light-isolating portion 14 can completely prevent the fluorescence used for temperature detection from entering the reaction chamber 12, completely preventing the fluorescence used for temperature detection from interfering with the detection results of the fluorescence detection device 6, and thus ensuring PCR detection accuracy. The light-isolating portion 14 can be optionally black or opaque, and preferably can be made of a metal material.

[0152] Optionally, the light-isolating portion 14 may be a black coating applied to the outside of the cavity wall 11. The specific material of the light-isolating portion 14 is not limited herein, as long as the light-isolating portion 14 is insoluble in the coolant, can be applied to the outside of the cavity wall 11, and has a heat-conducting function. The first fluorescent temperature-sensitive portion 13 is a layer of fluorescent temperature-sensitive material applied to the light-isolating portion 14.

[0153] like Figure 6b-Figure 7 As shown, specifically, the first fluorescent temperature-sensitive portion 13 is disposed on the outside of the first wall 111, and at least a portion of the second wall 112 is made of a 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 temperature through the first fluorescent temperature-sensitive portion 13 disposed on the first wall 111. The first wall 111 and the second wall 112 are disposed relative to each other, thereby further avoiding the mutual influence between temperature detection and detection of the content of the object to be detected. Of course, in other optional embodiments, at least a portion of the side wall 113 may also be made of a transparent material 114, and the fluorescence detection device 6 may detect through the transparent material 114 on the side wall 113.

[0154] like Figure 7 As shown, optionally, the first temperature detection mechanism 3 includes a first fluorescent receiving and transmitting component 33 and a first controller, wherein the first fluorescent receiving and transmitting 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, and the first controller is electrically connected to the first fluorescent receiving and transmitting component 33, and is used to convert the light intensity signal detected by the first fluorescent receiving and transmitting component 33 into a temperature signal.

[0155] It is understood that the first temperature detection mechanism 3 and the second temperature detection mechanism 4 have the same temperature measurement principle. The difference between the two is that the first temperature detection mechanism 3 does not need to extend into the reaction solution to detect the temperature. The first temperature detection mechanism 3 can perform temperature detection on one side of the first fluorescent temperature-sensitive portion 13. The first fluorescent temperature-sensitive portion 13 prepared in this embodiment 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 is configured to emit excitation light of a preset intensity. The first incident channel 31 is connected to the first light source and configured to transmit the excitation light from the first light source to the first fluorescence temperature-sensitive portion 13. The first receiving channel 32 is configured to transmit the excitation light at the first fluorescence temperature-sensitive portion 13 to the first detector 34. The first detector 34 is capable of detecting 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 output 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 fluorescent temperature-sensitive portion 13 to ensure the accuracy of the detection result. Furthermore, the first incident channel 31 and the first receiving channel 32 can be set adjacent to each other to further improve the detection accuracy. Figure 8 As shown, the first incident channel 31 and the first receiving channel 32 may not be specific physical objects, but are propagation channels of light in space.

[0158] Example 4

[0159] This embodiment provides a PCR amplification method, which can use the vector 1 provided in Example 1 and the PCR detection system provided in Example 2 to perform the PCR amplification method of this embodiment 4, or use the vector 1 provided in Example 1 and the PCR detection system provided in Example 3 to perform the PCR amplification method of this embodiment 4.

[0160] PCR amplification methods include:

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

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

[0163] The electromagnetic heating mechanism generates a magnetic field, causing the carrier 1 to heat up rapidly, thereby increasing the temperature of the reaction solution within the carrier 1. The cooling mechanism cools the carrier 1, thereby reducing the temperature of the reaction solution within the carrier 1. The electromagnetic heating mechanism and the cooling mechanism maintain the reaction solution at the denaturation temperature, extension temperature, and annealing temperature during the PCR reaction for a period of time. The coolant in the cooling mechanism continuously flows, allowing for ready cooling at any time, achieving rapid cooling.

[0164] 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. Therefore, even if condensed water is generated 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. Furthermore, because the electromagnetic heating mechanism and the carrier 1 can heat the reaction liquid in the carrier 1 without contact, the time required for heat conduction between the carrier 1 and the electromagnetic heating mechanism is saved when heating the carrier 1, thereby achieving rapid heating and accelerating the PCR reaction process and PCR reaction efficiency. This solves the problem of long detection time and slow heating and cooling speed caused by the large heat capacity of the heater and the large thermal resistance between the heater and the carrier 1.

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

[0166] The carrier 1 is in direct contact with the cooling liquid.

[0167] The carrier 1 is in direct contact with the coolant, eliminating other heat-conducting structures between the carrier 1 and the coolant, and can achieve rapid cooling of the carrier 1.

[0168] Optionally, the temperature of the carrier 1 can be adjusted by adjusting the power of the electromagnetic heating mechanism and / or by adjusting 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 reducing 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 reducing the duty cycle or increasing the current, and / or reducing the flow rate of the coolant.

[0169] The PCR detection method provided in this embodiment can achieve rapid heating and cooling. For example, compared with the traditional contact heater, the cooling mechanism needs to cool the heater and then cool the carrier 1 through the heater. The time required for intermediate heat conduction is long. The cooling mechanism in this embodiment does not need to cool the electromagnetic heating mechanism, but directly cools the carrier 1, so the cooling speed can be accelerated. For example, when the contact heater heats the carrier 1, the heat conduction between the carrier 1 and the heater takes a certain amount of time, and the heater itself also takes time to heat up. In this embodiment, the electromagnetic heating mechanism directly generates eddy currents in the carrier 1, and the eddy currents cause the carrier 1 to heat up, thereby heating the reaction liquid in the carrier 1, thereby greatly reducing the time required for heat conduction, thereby improving the heating efficiency of the reaction liquid. For example, the coolant flows continuously. During the PCR reaction process, there is no need to introduce coolants of different temperatures to achieve temperature rise and fall. This can greatly reduce the reaction time of the cooling mechanism and is conducive to achieving rapid temperature rise and fall.

[0170] PCR amplification methods also include:

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

[0172] Detecting the temperature of the reaction solution by using fluorescent temperature-sensitive materials and the intensity of the excitation light includes:

[0173] Continuously emitting excitation light to the fluorescent temperature-sensitive 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 liquid is obtained through the intensity of the collected excitation light.

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

[0177] The temperature detection mechanism emits a monochromatic excitation light with a wavelength of λ0. After the excitation light is irradiated to the fluorescent temperature-sensitive material, the fluorescent molecules with a wavelength of λ0 will produce light of another wavelength as the temperature of the fluorescent temperature-sensitive material changes, thereby causing the intensity of the excitation light with a wavelength of λ0 to weaken. The intensity I of the excitation light after the intensity is weakened is collected. The corresponding relationship between the intensity I and the temperature T is as follows: Figure 5 As shown, and satisfy the formula: The unit of T1 and T2 is K.

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

[0179] Although the present invention has been described in detail above using general explanations, specific embodiments, and experiments, it will be apparent to those skilled in the art that modifications and improvements may be made based on the present invention. Therefore, such modifications and improvements, which do not depart from the spirit of the present invention, are intended to be within the scope of protection claimed herein.

Claims

1. A carrier, characterized in that The invention comprises a cavity wall (11), wherein the cavity wall (11) encloses a reaction cavity (12) for accommodating a reaction liquid, and at least a portion of the cavity wall (11) can be heated by an electromagnetic field.

2. The carrier according to claim 1, characterized in that The carrier further includes a temperature sensing portion, and the temperature sensing portion is used to reflect the temperature of the reaction liquid.

3. The carrier according to claim 2, characterized in that The temperature sensing portion comprises a first fluorescent temperature-sensitive portion (13) arranged on the outside of the cavity wall (11), the first fluorescent temperature-sensitive portion (13) comprises a fluorescent temperature-sensitive material, and the first fluorescent temperature-sensitive portion (13) can reflect the temperature of the reaction liquid.

4. The carrier according to claim 3, characterized in that A light-isolating portion (14) is further provided between the first fluorescent temperature-sensitive portion (13) and the cavity wall (11).

5. The carrier according to any one of claims 2 to 4, characterized in that The cavity wall (11) comprises a first wall (111) and a second wall (112) arranged opposite to each other, and a side wall (113) arranged between the first wall (111) and the second wall (112); the temperature sensing portion is arranged on the outside of the first wall (111); and at least a portion of the second wall (112) or at least a portion of the side wall (113) is made of a transparent material (114).

6. The carrier according to claim 1, characterized in that At least part of the cavity wall (11) is formed by a heat conducting sheet (115), and the heat conducting sheet (115) can be heated by an electromagnetic field.

7. The carrier according to claim 6, characterized in that The heat conducting sheet (115) is a metal sheet.

8. The carrier according to claim 6 or 7, characterized in that The thickness of the heat conducting sheet (115) is 10-100 μm.

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

10. The carrier according to claim 9, characterized in that The transparent material (114) is polydimethylsiloxane, polypropylene, organic glass or polycarbonate.

11. The carrier according to any one of claims 1-4, 6-7, 9-10, characterized in that The reaction chamber (12) and / or the carrier are flat structures.

12. The carrier according to claim 11, characterized in that The flat structure means that the dimension of the reaction chamber (12) or the carrier perpendicular to its thickness direction is larger than its thickness direction.

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

1.

14. The carrier according to claim 13, characterized in that The size ratio is 50:1 to 100:

1.

15. A PCR detection system, characterized in that: It comprises an electromagnetic heating mechanism and a cooling mechanism, wherein the electromagnetic heating mechanism is used to heat the carrier according to any one of claims 1 to 14, and the cooling mechanism is used to cool the carrier according to any one of claims 1 to 14.

16. The PCR detection system according to claim 15, characterized in that The PCR detection system further comprises a temperature detection mechanism, which is used to detect the temperature of the reaction solution in the carrier.

17. The PCR detection system according to claim 16, characterized in that: The temperature detection mechanism is a first temperature detection mechanism (3), which detects the temperature of the reaction liquid by detecting the fluorescence intensity at the temperature sensing part of the carrier. The carrier is the carrier described in claims 2-4 and when claim 11 refers to any one of claims 2-3, the carrier described in any one of claims 10-13.

18. The PCR detection system according to claim 17, characterized in that The first temperature detection mechanism (3) comprises: A first fluorescent light receiving and emitting component (33) is used for emitting excitation light of a preset intensity to the temperature sensing portion and receiving and detecting the intensity of the excitation light at the temperature sensing portion; The first controller is electrically connected to the first fluorescent light receiving component (33) and is used to convert the light intensity signal detected by the first fluorescent light receiving component (33) into a temperature signal.

19. The PCR detection system according to claim 18, characterized in that The first fluorescent light receiving assembly (33) comprises: a first light source, configured to emit the excitation light of a preset intensity; a first incident channel (31), connected to the first light source and used to transmit the excitation light of the first light source to the temperature sensing portion; A first receiving channel (32) and a first detector (34), wherein the first receiving channel (32) is used to transmit the excitation light at the temperature sensing portion to the first detector (34), and the first detector (34) is capable of detecting the intensity of the excitation light transmitted by the first receiving channel (32).

20. The PCR detection system according to claim 16, characterized in that The temperature detection mechanism is a second temperature detection mechanism (4), which is used to detect the temperature of the reaction liquid in the carrier; the carrier is the carrier described in claims 1-7 and when claim 11 refers to any one of claims 6-7, 9-10, the carrier described in any one of claims 11-13.

21. The PCR detection system according to claim 20, characterized in that The second temperature detection mechanism (4) comprises: A second fluorescent temperature-sensitive portion (41) comprises a fluorescent temperature-sensitive material, and the second fluorescent temperature-sensitive portion (41) can be located in the reaction liquid in the reaction chamber (12) of the carrier; a second fluorescent light receiving and emitting component (45), the second fluorescent light receiving and emitting component (45) being used to emit excitation light of a preset intensity to the second fluorescent temperature-sensitive portion (41) and to receive and detect the intensity of the excitation light at the second fluorescent temperature-sensitive portion (41); The second controller is electrically connected to the second fluorescent light receiving component (45) and is used to convert the light intensity signal detected by the second fluorescent light receiving component (45) into a temperature signal.

22. The PCR detection system according to claim 21, characterized in that The second fluorescent light receiving assembly (45) comprises: a second incident channel (42), configured to transmit the excitation light to the second fluorescent temperature-sensitive portion (41); a second receiving channel (43), the second receiving channel (43) being used to transmit the excitation light at the second fluorescent temperature-sensitive portion (41); The second output end (421) of the second incident channel (42), the second incident end of the second receiving channel (43), and the second fluorescent temperature-sensitive portion (41) can all be located in the reaction liquid of the reaction chamber (12).

23. The PCR detection system according to claim 22, characterized in that The second temperature detection mechanism (4) further comprises a light-isolating covering portion (44), wherein at least the second output 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 portion (41) are all arranged in the light-isolating covering portion (44).

24. The PCR detection system according to claim 22, characterized in that The second fluorescent temperature-sensitive portion (41) comprises a fluorescent temperature-sensitive material coated on the second output end (421) of the second incident channel (42); and / or the second fluorescent temperature-sensitive portion (41) is connected to the second output end (421) of the second incident channel (42) and extends from the second incident channel (42).

25. The PCR detection system according to any one of claims 15 to 24, characterized in that: The cooling mechanism comprises a container (5), a cooling liquid continuously flows in the container (5), and the carrier can be inserted into the cooling liquid.

26. The PCR detection system according to claim 25, characterized in that The electromagnetic heating mechanism comprises a coil (2) for flowing electric current.

27. The PCR detection system according to claim 26, characterized in that The coil (2) is arranged on the upper side or the lower side of the container (5), or the coil (2) is sleeved on the outside of the container (5), or the coil (2) is arranged inside the container (5), and / or The PCR detection system further comprises a fluorescence detection device (6), and the fluorescence detection device (6) is used to detect the content of the substance to be detected in the reaction solution.

28. The PCR detection system according to claim 27, characterized in that The fluorescence detection device (6) is arranged outside the container (5).

29. The PCR detection system according to claim 27, characterized in that The fluorescence detection device (6) is located on one axial side or one radial side of the coil (2).

30. A PCR amplification method, characterized in that: The PCR amplification method is performed using the vector according to any one of claims 1 to 14 and the PCR detection system according to any one of claims 15 to 29, wherein the PCR amplification method comprises: The coolant flows continuously in the cooling mechanism to continuously cool the carrier; The electromagnetic heating mechanism generates a magnetic field and heats the carrier through the magnetic field.

31. The PCR amplification method according to claim 30, characterized in that By adjusting the power of the electromagnetic heating mechanism, and / or adjusting the temperature of the carrier by adjusting the temperature and / or flow rate of the coolant in the cooling mechanism; and / or The cooling liquid continuously flows in the cooling mechanism to continuously cool the carrier (1), comprising: The carrier is in direct contact with the coolant.

32. The PCR amplification method according to claim 30 or 31, characterized in that The PCR amplification method further comprises: The temperature of the reaction solution is detected by the fluorescent temperature-sensitive material and the intensity of the excitation light.

33. The PCR amplification method according to claim 32, characterized in that The method of detecting the temperature of the reaction solution by using a fluorescent temperature-sensitive material and the intensity of the excitation light comprises: continuously emitting excitation light toward the fluorescent temperature-sensitive material; 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 liquid is obtained according to the intensity of the collected excitation light.

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