A high-efficiency fluorescence quantitative PCR instrument

Abstract

The invention discloses a high-efficiency fluorescence quantitative PCR instrument, comprising an optical reader, a scanning device, a chamber, a heat cover, a PCR amplification temperature control component, a lifting component, a liquid cooling system and a power supply box; the invention has the advantages that: the new optical reader is suitable for collecting different types of fluorescence, can overcome the cross-interference of the detection light path, improve the fluorescence collection efficiency, and shorten the fluorescence collection time; the "bow"-shaped constant temperature water tank, the liquid cooling system, the independent air duct and the sealed heat cover can provide a stable PCR reaction environment and temperature.

Description

Technical Field

[0001] The invention relates to a PCR instrument, in particular to a high-efficiency fluorescence quantitative PCR instrument, belonging to the field of PCR instruments. Background Art

[0002] Real-time fluorescence quantitative PCR (Quantitative Real-time PCR) technology refers to adding fluorescent groups to the PCR reaction system, using the accumulation of fluorescent signals to monitor the entire PCR process in real time, and finally using the standard curve to perform quantitative analysis on unknown templates. As a commonly used analytical method in molecular biology, real-time fluorescence quantitative PCR technology has been widely used in scientific research and clinical practice.

[0003] The PCR-DNA / RNA real-time fluorescence quantitative detection system is generally composed of a real-time fluorescence quantitative PCR instrument, fluorescence quantitative reagents, a computer and automatic analysis software. The real-time fluorescence quantitative PCR instrument is used to monitor the fluorescence of the real-time fluorescence quantitative reagent and transmit the collected fluorescence data to the connected computer. The fluorescence data is displayed in the form of a chart through the real-time analysis software.

[0004] Real-time fluorescence quantitative PCR instruments generally include multiple detection light paths, which can simultaneously perform multiple PCR and allele, SNP and other analyses, and obtain the required test results through a single experiment. Each detection light path of a real-time fluorescence quantitative PCR instrument generally consists of an excitation light generator, a filter, an excitation end lens, a dichroic mirror, a PCR reaction tube end lens, a collection end filter, a receiving lens and a detection device.

[0005] There are two ways to implement a real-time fluorescence quantitative PCR instrument with multiple detection light path structures:

[0006] The first is that the receiving lens and the detection structure are two separate channels. The disadvantage is that the detection structure will have differences between devices, resulting in deviations in the analysis results and increasing economic costs;

[0007] The second is to use a common excitation light source or detection device, because the devices are the same, there will be no device differences. But the disadvantage is: optical path difference and cross-interference between optical paths are easy to occur between optical lenses and structures. The reason is that because the filter adopts the imaging method of the wheel-gating structure, the optical path difference between the 96 wells on the 96-well plate and the excitation light source and the detection device is different, resulting in different excitation and receiving efficiencies, and the detection results will be affected to varying degrees. In addition, because the spectrum of commonly used QPCR dyes is concentrated in 500-700nm, when 5-6 optical channels are integrated simultaneously in this range, it is inevitable that the fluorescence collection filter spectrum of the low-band optical channel will intersect with the excitation filter spectrum of the adjacent high-band optical channel, which is the cross-interference of the optical path.

[0008] When this intersection phenomenon occurs, in theory, the excitation light of each optical channel will be cut off by the fluorescence collection filter corresponding to its channel, and will not be collected by the detection device. However, in actual application experiments, when the excitation light generator excites the target PCR reaction tube to be tested, the excitation light will crosstalk through the tube wall of the target tube to be tested to its adjacent PCR reaction tube, that is, the crosstalk tube. If the top cover of the crosstalk tube is facing another collection light path at this time, then the excitation light that crosstalks through the target tube to be tested will be received by the same detection device through the fluorescence collection filter of the corresponding light path on the crosstalk tube, thereby causing optical channel crosstalk.

[0009] In addition, in actual use, the existing real-time fluorescence quantitative PCR instrument will have the following technical problems in addition to the above technical problems:

[0010] 1. During the PCR gene amplification process, due to the rapid change of ambient temperature, the reaction reagents will inevitably evaporate, which greatly affects the normal progress of the reaction and the determination of the results.

[0011] 2. The long-term operation of the real-time fluorescence quantitative PCR instrument will cause the body to overheat, affecting the normal analysis work.

[0012] 3. The scanning time is too slow and the scanning speed is too slow; high-resolution melting curve is not supported. Summary of the invention

[0013] In order to solve the defects of the prior art, the present invention provides a high-efficiency fluorescence quantitative PCR instrument, which is suitable for collecting different types of fluorescence, can overcome the cross-interference of the detection light path, improve the fluorescence collection efficiency, and can provide a stable PCR reaction environment and temperature.

[0014] The technical solution of the present invention is:

[0015] A high-efficiency fluorescence quantitative PCR instrument, comprising an optical reader, a scanning motion component, a chamber, a hot cover, a PCR amplification temperature control component, a lifting motion component, a liquid cooling system and a power box;

[0016] The optical reader is composed of at least one optical detection channel; each optical detection channel includes: an excitation light source, a first filter, a first convex lens, a reflector, a dichroic mirror, a second convex lens, a third convex lens, a second filter and a detection device; the light source irradiated by the excitation light source passes through the first filter and the first convex lens in sequence and then directly hits the reflector, and then the light path direction is reflected on the dichroic mirror, and the dichroic mirror converts the light path direction and vertically shoots it through the second convex lens to the target tube to be detected PCR reaction tube; the fluorescence excited from the target tube to be detected PCR reaction tube passes through the second convex lens, the dichroic mirror, the third convex lens and the second filter in sequence and then enters the detection device for detection.

[0017] Furthermore, the number of the optical detection channels is multiple; each fluorescence corresponds to at least two optical detection channels;

[0018] Furthermore, the excitation light source is an LED, preferably a monochromatic LED.

[0019] Furthermore, the detection device is PD, APD, PMT, MPPC, etc., preferably MPPC.

[0020] The scanning motion assembly is used to install the optical reader and drive the optical reader to scan along the horizontal and vertical axes on a two-dimensional plane; specifically, the optical reader is controlled by the vertical axis motor to move along the horizontal axis on the two-dimensional plane; and is controlled by the horizontal axis motor to move along the vertical axis; the scanning motion assembly is composed of a horizontal scanning motion assembly and a vertical scanning motion assembly; the horizontal scanning motion assembly is used to drive the vertical scanning motion assembly to move horizontally; the horizontal scanning motion assembly includes a horizontal axis control motor, a horizontal axis transmission belt and a horizontal axis guide rail; the horizontal axis control motor drives the horizontal axis transmission belt to transmit; the number of the horizontal axis guide rails is 2, namely the front horizontal axis guide rail and the rear horizontal axis guide rail; The longitudinal scanning motion component comprises a longitudinal axis control motor, a longitudinal axis transmission belt, a longitudinal axis guide rail, a front sliding member and a rear sliding member; the longitudinal axis control motor drives the longitudinal axis transmission belt to transmit; the longitudinal axis control motor is installed on the rear sliding member; the longitudinal axis transmission belt drives the optical reading head to move longitudinally; the front and rear ends of the longitudinal axis guide rail are respectively installed on the front sliding member and the rear sliding member; the front sliding member is installed on the front transverse axis guide rail; the rear part of the rear sliding member is a synchronous belt connector, and the synchronous belt connector is a clip structure, which is used to fix the rear sliding member on the transverse axis transmission belt, thereby driving the longitudinal scanning motion component to move transversely;

[0021] Furthermore, the scanning motion assembly further comprises a connecting plate, the connecting plate being used to mount the optical reading head; a fixing clip is provided at the tail of the connecting plate, which is used to connect the longitudinal axis transmission belt of the scanning motion assembly; a slide rail is also provided on the connecting plate, which is used to connect the longitudinal axis guide rail 225 and move on the longitudinal axis guide rail;

[0022] Furthermore, the high-efficiency fluorescence quantitative PCR instrument is also provided with a chamber body, which is used to install an optical reader, a scanning motion component, a thermal cover, a PCR amplification temperature control component, a lifting motion component, a liquid cooling system and a power supply box;

[0023] An air outlet, a heat dissipation port and a power interface are provided at the rear of the warehouse body; the air outlet is used for heat dissipation; a fan is installed inside the air outlet for heat dissipation inside the warehouse body; the heat dissipation port is used for heat dissipation by a radiator; a radiator is installed inside the heat dissipation port; the power interface is used for connecting an external power cord; a warehouse opening is provided at the top of the warehouse body, and the warehouse opening is provided with a warehouse door; front and rear air inlets are provided at the bottom of the warehouse body; the front air inlet corresponds to the air outlet, and the heat inside the warehouse body is dissipated by the fan; the rear air inlet corresponds to the heat dissipation port, and the heat dissipation of the radiator is dissipated by the heat dissipation fan; a horizontal fixed plate is provided inside the warehouse body; the fixed plate is used for installing a scanning motion component and a heat cover; a sampling port is provided at the front of the fixed plate, corresponding to the warehouse opening, and is used to take and place PCR reaction tubes; guide rails are provided on the left and right sides of the bottom of the sampling port of the fixed plate, and the guide rails are used to slide the heat cover so that the heat cover can be extended and retracted forward and backward;

[0024] Furthermore, the front and rear parts of the warehouse body are respectively provided with data ports for connecting external devices;

[0025] The heat cover is located directly above the PCR amplification temperature control component and extends forward and backward along the fixed plate sampling port; the heat cover can provide a high temperature of 100°C to prevent the PCR system in the PCR reaction tube from gasifying and evaporating, improve the fluorescence collection efficiency, and enhance the detection sensitivity; the length and width of the heat cover are consistent with the length and width of the sealing structure; sliders are installed on both sides of the heat cover for sliding on the guide rails of the fixed plate sampling port; the heat cover cover plate is provided with light holes corresponding to the bottom thermal cycle module one by one; a lens is provided in the light hole, and the lens can be selected as a convex lens or a plane mirror according to actual conditions, in order to improve the fluorescence collection efficiency; the optical reader receives the fluorescence excited by the PCR reaction tube system on the lower thermal cycle module through the light hole of the heat cover, and the fluorescence is received by the optical reader through the lens in the light hole at the top of the PCR reaction tube cover.

[0026] Furthermore, a sealing ring is installed on the thermal cover to assist sealing;

[0027] The PCR amplification temperature control assembly includes a base, a sealing structure, a thermal cycle module, a semiconductor refrigerator, a constant temperature water tank and a high-low tube adaptive structure; the base is used to install and fix the sealing structure, the thermal cycle module, the semiconductor refrigerator and the constant temperature water tank; the top of the base is a mounting plate, and the middle of the mounting plate is provided with a mounting port, and the mounting port is used to install and fix the thermal cycle module; the four corners of the base are respectively installed with anti-collision rods, and the height of the anti-collision rods is slightly higher than the height of the thermal cycle module, so as to prevent the fixed plate from colliding with the thermal cycle module after the PCR reaction tube is taken out;

[0028] The sealing structure is made of silicone and is used for sealing; the sealing structure is an elliptical structure; the bottom of the sealing structure is designed to be open and is installed on the inner edge of the installation opening of the base; the sealing structure and the thermal cover are fitted in time to achieve a sealing effect and form a sealed space, so that the PCR reaction tube carried by the thermal cycle module in the sealing joint forms a stable PCR reaction environment to prevent external interference;

[0029] The thermal cycle module is used to carry PCR reaction tubes. The thermal cycle module has the same size and specifications as a conventional 96-well plate and is compatible with a single PCR reaction tube, 8-row tubes and a 96-well plate. The semiconductor refrigerator is installed at the bottom of the thermal cycle module. The semiconductor refrigerator is used to provide various temperatures required for PCR reactions in PCR reaction tubes in the thermal cycle module. A constant temperature water tank is installed at the bottom of the semiconductor refrigerator. The constant temperature water tank is used for heat dissipation. The constant temperature water tank is a rectangular heat dissipation block, and heat is dissipated by the flow of refrigerant in the rectangular heat dissipation block. A double "bow"-shaped channel is arranged inside the constant temperature water tank, and the inlet and outlet of the double "bow"-shaped channel are symmetrical on the left and right. The double "bow"-shaped channel consists of two "bow"-shaped channels connected at the bottom in opposite directions. The channels are staggered with each other in order to reduce the temperature difference of the refrigerant passing through between the inlet and the outlet and improve the uniformity of the temperature of the constant temperature water tank. Specifically, when the refrigerant enters the channel from the inlet of the double "bow"-shaped channel, the temperature of the refrigerant is relatively low. When the refrigerant passes through the channel, the heat emitted by the PCR amplification temperature control component is cooled, thereby causing the temperature of the refrigerant to rise, thereby achieving the purpose of heat dissipation. The double "bow"-shaped channels are designed with the "bow"-shaped channels staggered with each other. While cooling the heat emitted by the PCR amplification temperature control component, the newly entered refrigerant can continue to cool the refrigerant that has entered the adjacent pipe, thereby reducing the temperature difference of the refrigerant in the channel between the inlet and the outlet and improving the uniformity of the temperature of the constant temperature water tank 55.

[0030] Furthermore, a liquid inlet and a liquid outlet are arranged in parallel on one side of the bottom of the constant temperature water tank, which are respectively connected to the inlet and outlet of the double "bow" type channel; the liquid inlet is connected to the water pump through a pipe; and the liquid outlet is connected to the radiator inlet through a pipe.

[0031] The high-low tube adaptive structure is used to adjust the height of the PCR reaction tube in the thermal cycle module; the high-low tube adaptive structure is composed of a support plate and a lifting plate; the support plate is installed at the bottom of the constant temperature water tank; guide columns are provided at the four corners of the support plate; springs are installed outside the guide columns to play a self-adaptive role; the lifting plate is installed on the top of the scissor-type lifting platform; through-type guide holes are provided at the four corners of the lifting plate surface, and the guide holes correspond to the guide columns respectively; the guide columns can enter the guide holes;

[0032] The principle of the high-low tube adaptive structure is as follows: when the PCR reaction tube is placed in the thermal cycle module, the thermal cover is closed and buckled on the thermal cycle module; the scissor-type lifting platform pushes the lifting plate to rise upward, thereby driving the support plate, the constant temperature water tank and the semiconductor refrigerator to rise together; when the PCR reaction tube in the thermal cycle module reaches the bottom of the thermal cover, the lifting plate continues to rise, and at this time the guide column on the support plate continues to penetrate into the guide hole of the lifting plate, causing the spring on the guide column to be continuously compressed; the lifting plate adjusts the elastic force of the spring by compression, so that the PCR reaction tube in the thermal cycle module can fit more closely with the bottom of the thermal cover;

[0033] The lifting motion component is a scissor-type lifting platform, and the lifting and lowering of the scissor-type lifting platform is controlled by a lifting motor; the lifting motion component drives the PCR amplification temperature control component to rise and fall; when placing or removing the PCR reaction tube, the lifting motion component vertically lifts the PCR amplification temperature control component to the upper part of the sampling port, which is convenient for taking and placing the PCR reaction tube, and also prevents the PCR reaction tube from falling into the warehouse.

[0034] The liquid cooling system is used for heat dissipation, and the liquid cooling system consists of a water pump, a liquid storage tank and a radiator; the water pump is used to extract the refrigerant in the liquid storage tank and input it into the constant temperature water tank through a pipeline; the water pump outlet is connected to the liquid inlet of the constant temperature water tank through a pipeline; the water pump liquid extraction port is connected to the liquid storage tank outlet; the liquid storage tank is used to store refrigerant; the liquid storage tank inlet is connected to the heat pipe outlet of the radiator; the radiator is used to dissipate heat for the refluxed refrigerant, and the radiator has a built-in radiator-shaped heat pipe; the heat pipe inlet is connected to the liquid outlet of the constant temperature water tank through a pipeline, and is used to cool the refluxed refrigerant; a number of heat sinks are provided on the heat pipe body.

[0035] Furthermore, a cooling fan is installed on the radiator to dissipate heat for the radiating pipe;

[0036] Furthermore, the liquid storage tank, the radiator and a plurality of partitions form an independent chamber, the bottom of the chamber is a rear air inlet, one side is a heat dissipation port, and is isolated from other internal structures of the instrument; the purpose of the independent chamber is to form an independent external circulation heat dissipation air duct to avoid interfering with the temperature in the chamber; at the same time, it also reduces the problem of dust falling inside the instrument;

[0037] Furthermore, a power supply box is installed in the chamber of the high-efficiency fluorescent quantitative PCR instrument; the power supply box is used to supply power to the device and reasonably distribute electrical energy.

[0038] The beneficial effects of the present invention are:

[0039] 1. By setting up multiple optical detection channels, the detection efficiency is improved, and different fluorescence is applicable. The optical path position setting effectively avoids cross interference between optical paths;

[0040] 2. During the PCR gene amplification process, the sealed structure design and PCR amplification temperature control components make the change of environmental temperature controllable, and the evaporation of reaction reagents is avoided;

[0041] 3. Due to the addition of a liquid cooling system, the real-time fluorescence quantitative PCR instrument reduces overheating when working for a long time, thereby improving the durability and stability of the equipment.

[0042] The present invention will be further described below in conjunction with the accompanying drawings and embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 The internal structure of a high-efficiency fluorescence quantitative PCR instrument according to an embodiment of the present invention (I) is an overall schematic diagram;

[0044] Figure 2 Schematic diagram of the internal structure of a high-efficiency fluorescence quantitative PCR instrument according to an embodiment of the present invention (II);

[0045] Figure 3 This is a schematic diagram of the left side cross-sectional structure of an optical reader of a high-efficiency fluorescence quantitative PCR instrument according to an embodiment of the present invention;

[0046] Figure 4 This is a schematic diagram of the working principle of the optical reader of the present invention;

[0047] Figure 5 This is a schematic diagram of the structure of a high-efficiency fluorescence quantitative PCR instrument according to an embodiment of the present invention;

[0048] Figure 6 This is a schematic diagram of the rear structure of a chamber body of a high-efficiency fluorescence quantitative PCR instrument according to an embodiment of the present invention;

[0049] Figure 7A schematic diagram of the structure of a PCR amplification temperature control component of a high-efficiency fluorescence quantitative PCR instrument according to an embodiment of the present invention;

[0050] Figure 8 A schematic diagram of a PCR amplification temperature control component and a lifting structure of a high-efficiency fluorescence quantitative PCR instrument according to an embodiment of the present invention;

[0051] Fig. 9 A schematic cross-sectional structure diagram of a PCR amplification temperature control component of a high-efficiency fluorescence quantitative PCR instrument according to an embodiment of the present invention;

[0052] Fig.10 A schematic diagram of the internal structure of a constant temperature water tank of a high-efficiency fluorescence quantitative PCR instrument according to an embodiment of the present invention;

[0053] In the figure: 1 optical reader, 11 excitation light source, 12 first filter, 13 first convex lens, 14 reflector, 15 dichroic mirror, 16 second convex lens, 17 third convex lens, 18 second filter, 19 detection device; 2 scanning motion assembly, 21 horizontal scanning motion assembly, 211 horizontal axis control motor, 212 horizontal axis transmission belt, 213 front horizontal axis guide rail, 214 rear horizontal axis guide rail, 22 longitudinal scanning motion assembly, 221 longitudinal axis control motor, 222 longitudinal axis transmission belt, 223 connecting plate, 224 slide rail, 225 longitudinal axis guide rail, 226 front sliding member, 227 rear sliding member, 3 warehouse body, 31 function key, 32 indicator light, 33 air outlet, 331 fan, 34 heat dissipation port, 35 power interface, 36 warehouse port, 361 warehouse door, 37 air inlet, 38 fixing plate, 381 sampling port, 39 data port, 4 heat cover, 41 light hole, 5 PCR amplification temperature control component, 51 base, 511 mounting plate, 512 anti-collision rod, 52 sealing structure, 53 thermal cycle module, 531 PRC reaction tube, 54 semiconductor refrigerator, 55 constant temperature water tank; 551 double "bow" type channel, 552 liquid inlet, 553 liquid outlet, 56 high and low tube adaptive structure, 561 support plate, 562 guide column, 563 lifting plate, 564 guide hole, 6 lifting motion component, 61 lifting motor, 7 liquid cooling system; 71 water pump, 72 liquid storage tank, 73 radiator, 731 heat pipe, 732 cooling fan, 8 power box. DETAILED DESCRIPTION

[0054] The preferred embodiments of the present invention are described below. It should be understood that the preferred embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention. Example

[0055] like Figure 1-9 As shown, it includes an optical reader 1, a scanning motion component 2, a chamber body 3, a thermal cover 4, a PCR amplification temperature control component 5, a lifting motion component 6, a liquid cooling system 7 and a power supply box 8;

[0056] The optical reader 1 is composed of at least one optical detection channel; each optical detection channel includes: an excitation light source 11, a first filter 12, a first convex lens 13, a reflector 14, a dichroic mirror 15, a second convex lens 16, a third convex lens 17, a second filter 18 and a detection device 19; the light source irradiated by the excitation light source 11 passes through the first filter 12 and the first convex lens 13 in sequence and then directly hits the reflector 14, and then the light path direction is reflected on the dichroic mirror 15, and the dichroic mirror 15 converts the light path direction and vertically shoots it to the target tube to be detected PCR reaction tube through the second convex lens 16; the fluorescence excited from the target tube to be detected PCR reaction tube passes through the second convex lens 16, the dichroic mirror 15, the third convex lens 17 and the second filter 18 in sequence and then enters the detection device 19 for detection.

[0057] Furthermore, the number of the optical detection channels is multiple; each fluorescence corresponds to at least two optical detection channels;

[0058] Furthermore, the excitation light source 11 is an LED, preferably a monochromatic LED.

[0059] Furthermore, the detection device 19 is PD, APD, PMT, MPPC, etc., preferably MPPC.

[0060] The scanning motion component 2 is used to install the optical reading head 1 and drive the optical reading head 1 to scan along the horizontal and vertical axis directions on a two-dimensional plane; specifically, the optical reading head 1 is controlled by the vertical axis motor to move along the horizontal axis on the two-dimensional plane; and is controlled by the horizontal axis motor to move along the vertical axis; the scanning motion component 2 is composed of a horizontal scanning motion component 21 and a vertical scanning motion component 22; the horizontal scanning motion component 21 is used to drive the vertical scanning motion component 22 to move horizontally; the horizontal scanning motion component 21 includes a horizontal axis control motor 211, a horizontal axis transmission belt 212 and a horizontal axis guide rail; the horizontal axis control motor 211 drives the horizontal axis transmission belt 212 to transmit; the number of the horizontal axis guide rails is 2, namely the front horizontal axis guide rail 213 and the rear horizontal axis guide rail 214; the vertical scanning motion component 22 includes a vertical axis control motor 211, a horizontal axis transmission belt 212 and a horizontal axis guide rail; The control motor 221, the longitudinal axis transmission belt 222, the longitudinal axis guide rail 225, the front sliding member 226 and the rear sliding member 227 are composed; the longitudinal axis control motor 221 drives the longitudinal axis transmission belt 222 to transmit; the longitudinal axis control motor 221 is installed on the rear sliding member 227; the longitudinal axis transmission belt 222 drives the optical reading head 1 to move longitudinally; the front and rear ends of the longitudinal axis guide rail 225 are respectively installed on the front sliding member 226 and the rear sliding member 227; the front sliding member 226 is installed on the front transverse axis guide rail 213; the rear sliding member 227 is installed on the rear transverse axis guide rail 214; the rear part of the rear sliding member 227 is a synchronous belt connector, and the synchronous belt connector is a clip structure, which is used to fix the rear sliding member 227 on the transverse axis transmission belt 212, thereby driving the longitudinal scanning motion component 22 to move transversely;

[0061] Furthermore, the scanning motion assembly 2 further includes a connecting plate 223, and the connecting plate 223 is used to install the optical reading head 1; a fixing clip is provided at the tail of the connecting plate 223, which is used to connect the longitudinal axis transmission belt of the scanning motion assembly; a slide rail 224 is also provided on the connecting plate 223, which is used to connect the longitudinal axis guide rail 225 and move on the longitudinal axis guide rail 225;

[0062] Furthermore, the efficient fluorescence quantitative PCR instrument is also provided with a chamber body 3, and the chamber body 3 is used to install the optical reader 1, the scanning motion component 2, the thermal cover 4, the PCR amplification temperature control component 5, the lifting motion component 6, the liquid cooling system 7 and the power supply box 8;

[0063] The front of the warehouse body 3 is provided with a function key 31 and an indicator light 32; the rear of the warehouse body 3 is provided with an air outlet 33, a heat dissipation port 34 and a power interface 35; the air outlet 33 is used for heat dissipation; a fan 331 is installed inside the air outlet 33 for heat dissipation inside the warehouse body 3; the heat dissipation port 34 is used for heat dissipation by a radiator 73; a radiator 73 is installed inside the heat dissipation port 34; the power interface 35 is used for connecting an external power cord; a warehouse opening 36 is provided on the top of the warehouse body 3, and the warehouse opening is provided with a warehouse door 361; the bottom of the warehouse body is provided with front and rear air inlets 37 ; The front air inlet corresponds to the air outlet, and the fan is used to dissipate heat inside the warehouse; the rear air inlet corresponds to the heat dissipation port, and the heat dissipation fan is used to dissipate heat from the radiator; a horizontal fixing plate 38 is provided inside the warehouse; the fixing plate 38 is used to install the scanning motion component 2 and the heat cover 4; a sampling port 381 is provided at the front of the fixing plate 38, corresponding to the warehouse opening 36, and is used to take and place PCR reaction tubes; guide rails are provided on the left and right sides of the bottom of the fixing plate sampling port 381, and the guide rails are used to slide the heat cover 4 so that the heat cover 4 can be extended forward and backward;

[0064] Furthermore, the front and rear parts of the compartment body 3 are respectively provided with data ports 39 for connecting external devices;

[0065] The heat cover 4 is located directly above the PCR amplification temperature control component 5, and is telescopic forward and backward along the sampling port 381 of the fixed plate 38; the heat cover 4 can provide a high temperature of 100°C to prevent the PCR system in the PCR reaction tube from gasifying and evaporating, thereby improving the fluorescence collection efficiency and the detection sensitivity; the length and width of the heat cover 4 are consistent with the length and width of the sealing structure; sliders are installed on both sides of the heat cover 4 for the purpose of sliding on the guide rails of the sampling port of the fixed plate; the cover plate of the heat cover 4 is provided with light holes 41 corresponding to the bottom thermal cycle modules one by one; a lens is provided in the light hole 41, and the lens can be selected as a convex lens or a plane mirror according to actual conditions, in order to improve the fluorescence collection efficiency; the optical reader 1 receives the fluorescence excited by the PCR reaction tube system on the lower thermal cycle module 53 through the light hole 41 of the heat cover 4, and the fluorescence is received by the optical reader through the lens in the light hole 41 at the top of the PCR reaction tube cover.

[0066] Furthermore, a sealing ring is installed on the thermal cover 4 to assist in sealing;

[0067] The PCR amplification temperature control component 5 is composed of a base 51, a sealing structure 52, a thermal cycle module 53, a semiconductor refrigerator 54, a constant temperature water tank 55 and a high-low tube adaptive structure 56; the base 51 is used to install and fix the sealing structure 52, the thermal cycle module 53, the semiconductor refrigerator 54 and the constant temperature water tank 55; the top of the base 51 is a mounting plate 511, and a mounting opening is provided in the middle of the mounting plate 511, and the mounting opening is used to install and fix the thermal cycle module 53; the four corners of the base 51 are respectively installed with anti-collision rods 512, and the height of the anti-collision rods 512 is slightly higher than the height of the thermal cycle module 53, and the purpose is to prevent the fixing plate 38 from colliding with the thermal cycle module 53 after the PCR reaction tube is taken out;

[0068] The sealing structure 52 is made of silicone and is used for sealing. The sealing structure 52 is an elliptical structure. The bottom of the sealing structure 52 is designed to be open and is installed on the inner edge of the installation opening of the base 51. When the instrument starts working, the sealing structure 52 and the thermal cover 4 are properly fitted to achieve a sealing effect, so that the PCR reaction tube 531 carried by the thermal cycle module 53 in the sealing joint forms a stable PCR reaction environment to prevent external interference.

[0069] The thermal cycle module 53 is used to carry PCR reaction tubes. The thermal cycle module 53 has the same size and specifications as a conventional 96-well plate and is compatible with a single PCR reaction tube 531, 8-row tubes and a 96-well plate. The semiconductor refrigerator 54 is installed at the bottom of the thermal cycle module 53. The semiconductor refrigerator 54 is used to provide various temperatures required for PCR reactions in the PCR reaction tubes in the thermal cycle module 53. A constant temperature water tank is installed at the bottom of the Par 54 sticker. The constant temperature water tank 55 is used for heat dissipation. The constant temperature water tank 55 is a rectangular heat dissipation block, which dissipates heat by the flow of refrigerant in the rectangular heat dissipation block. A double "bow"-shaped channel 551 is arranged inside the constant temperature water tank 55, and the inlet and outlet of the double "bow"-shaped channel 551 are symmetrical on the left and right. The double "bow"-shaped channel 551 is connected in two directions. The "bow"-shaped channels connected at the bottom are staggered with each other, the purpose of which is to reduce the temperature difference of the refrigerant passing through between the inlet and the outlet and improve the temperature uniformity of the constant temperature water tank; specifically, when the refrigerant enters the channel from the inlet of the double "bow"-shaped channel 551, the temperature of the refrigerant is relatively low. When the refrigerant passes through the channel, it will cool the heat emitted by the PCR amplification temperature control component 5, thereby causing the temperature of the refrigerant to increase, thereby achieving the purpose of heat dissipation. The double "bow"-shaped channels 551 are designed with "bow"-shaped channels staggered with each other, and the newly entered refrigerant can continue to cool the refrigerant that has entered the adjacent pipe while cooling the heat emitted by the PCR amplification temperature control component, thereby reducing the temperature difference of the refrigerant in the channel between the inlet and the outlet and improving the temperature uniformity of the constant temperature water tank 55.

[0070] Furthermore, a liquid inlet 552 and a liquid outlet 553 are arranged in parallel on one side of the bottom of the constant temperature water tank 55, which are respectively connected to the inlet and outlet of the double "bow"-shaped channel 551; the liquid inlet 552 is connected to the water pump through a pipe; the liquid outlet 553 is connected to the radiator inlet through a pipe.

[0071] The high-low tube adaptive structure 56 is used to adjust the height of the PCR reaction tube in the thermal cycle module 53; the high-low tube adaptive structure 56 is composed of a support plate 561 and a lifting plate 563; the support plate 561 is installed at the bottom of the constant temperature water tank 55; the four corners of the support plate 561 are provided with guide columns 562; the guide columns 562 are externally installed with springs to play a self-adaptive role; the lifting plate 563 is installed on the top of the scissor-type lifting platform; the four corners of the surface of the lifting plate 563 are provided with through-type guide holes 564, and the guide holes 564 correspond to the guide columns 562 respectively; the guide columns 562 can enter the guide holes 564;

[0072] The principle of the high-low tube adaptive structure 56 is as follows: after the PCR reaction tube is placed in the thermal cycle module 53, the thermal cover 4 is closed and buckled on the thermal cycle module 53; the scissor-type lifting platform pushes the lifting plate 563 to rise, thereby driving the support plate 561, the constant temperature water tank 55 and the semiconductor refrigerator 54 to rise together; when the PCR reaction tube in the thermal cycle module 53 reaches the bottom of the thermal cover 4, the lifting plate 563 continues to rise, and at this time, the guide column 562 on the support plate 561 continues to penetrate into the guide hole 564 of the lifting plate 563, causing the spring on the guide column 562 to be continuously compressed; the lifting plate 563 adjusts the elastic force of the spring by compression, so that the PCR reaction tube in the thermal cycle module 53 can fit more closely with the bottom of the thermal cover 4;

[0073] The lifting motion component 6 is a scissor-type lifting platform, and the lifting and lowering of the scissor-type lifting platform is controlled by a lifting motor 61; the lifting motion component 6 drives the PCR amplification temperature control component 5 to rise and fall; when placing or removing the PCR reaction tube 531, the lifting motion component 6 vertically lifts the PCR amplification temperature control component 5 to the upper part of the sampling port 381, which is convenient for taking and placing the PCR reaction tube 531, and also prevents the PCR reaction tube 531 from falling into the warehouse.

[0074] The liquid cooling system 7 is used for heat dissipation, and the liquid cooling system consists of a water pump 71, a liquid storage tank 72 and a radiator 73; the water pump 71 is used to extract the refrigerant in the liquid storage tank 72 and input it into the constant temperature water tank 55 through a pipeline; the outlet of the water pump 71 is connected to the liquid inlet of the constant temperature water tank 55 through a pipeline; the liquid extraction port of the water pump 71 is connected to the outlet of the liquid storage tank 72; the liquid storage tank 72 is used to store refrigerant; the inlet of the liquid storage tank 72 is connected to the outlet of the heat pipe 731 of the radiator 73; the radiator 73 is used to dissipate heat for the refluxed refrigerant, and the radiator 73 has a built-in heat pipe 731 in the shape of a radiator; the inlet of the heat pipe 731 is connected to the liquid outlet of the constant temperature water tank 55 through a pipeline, for cooling the refluxed refrigerant; the heat pipe 731 is provided with a plurality of heat sinks.

[0075] Furthermore, a cooling fan 732 is installed on the radiator 73 to dissipate heat for the heat pipe 731;

[0076] Furthermore, the liquid storage tank 72, the radiator 73 and a plurality of partitions form an independent chamber, the bottom of the chamber is a rear air inlet, one side is a heat dissipation port 34, and is isolated from other internal structures of the instrument; the purpose of the independent chamber is to form an independent external circulation heat dissipation air duct to avoid interfering with the temperature in the chamber; at the same time, it also reduces the problem of dust falling inside the instrument;

[0077] Furthermore, a power supply box 8 is installed in the chamber of the high-efficiency fluorescence quantitative PCR instrument; the power supply box 8 is used to supply power to the device and reasonably distribute electrical energy.

[0078] The working principle of the liquid cooling system is: the water pump draws out the refrigerant liquid in the liquid storage tank, and the refrigerant liquid flows into the constant temperature water tank along the pipeline and exchanges heat with the PCR amplification temperature control component; the refrigerant liquid after heat exchange passes through the constant temperature water tank and then enters the radiator along the pipeline for heat dissipation, and then flows back into the liquid storage tank to complete the refrigeration working cycle.

[0079] The workflow of the present invention is as follows:

[0080] Start the instrument, push open the door, and the hot cover moves backward. At the same time, the lifting motor drives the scissor-type lifting platform to drive the PCR amplification temperature control component to move vertically out of the warehouse. Place the PCR reaction tube in the thermal cycle module, and the scissor-type lifting platform enters the warehouse vertically. After entering the warehouse, the hot cover moves forward to the top of the sealing structure and the thermal cycle module; the scissor-type lifting platform controls the high and low tube adaptive structure to rise so that the PCR reaction tube in the thermal cycle module and the hot cover are adaptively crimped, so that the hot cover and the thermal cycle module are fully crimped; at this time, start the PCR amplification and detection process. On the two-dimensional plane, the scanning motion component drives the optical reader along the horizontal and vertical axes to cooperate with the PCR reaction process to detect the fluorescence in the PCR reaction tube; at the same time, the heat dissipation system drives the flow of refrigerant through the water pump to dissipate heat for the instrument.

[0081] When fluorescence detection is performed, when each optical detection channel on the optical reader passes through the PCR reaction tube position on the thermal cycle module corresponding to the hole in the thermal cover, the excitation light sources of all optical detection channels are lit at the same time, thereby stimulating the fluorescent dye in the system in the PCR reaction tube, and the emitted fluorescence is received by the MPPC after passing through the respective convex lenses and filters; since the LEDs of each optical path will light up at the same time, the device will determine the position of the PCR reaction tube of the optical reader in the thermal cycle module according to the current position feedback of each optical path, and determine the collected fluorescence signal from which optical path according to the lit LED, and then store the signal, draw a real-time fluorescence curve after analysis, and display the results and conduct subsequent analysis after the experiment.

[0082] In addition, in the description of the embodiments of the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, and it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0083] In the description of the present invention, it should be noted that the terms "center", "up", "down", "left", "right", "vertical", "horizontal", "inside", "outside" and the like indicate positions or positional relationships based on the positions or positional relationships shown in the accompanying drawings, which are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation of the present invention. In the several embodiments provided in the present application, it should be understood that the disclosed device can be implemented in other ways. The device embodiments described above are merely schematic. For example, the division of the units is only a logical function division. There may be other division methods in actual implementation. For example, multiple units or components may be combined or integrated into another system, or some features may be ignored or not executed. Another point is that the mutual coupling or direct coupling or communication connection shown or discussed may be through some communication interfaces, and the indirect coupling or communication connection of the device or unit may be electrical, mechanical or other forms.

Claims

1. A highly efficient fluorescence quantitative PCR instrument, characterized in that: It includes optical reader, scanning motion component, chamber, heat cover, PCR amplification temperature control component, lifting motion component, liquid cooling system and power box; The optical reader is composed of at least one optical detection channel; each optical detection channel includes: an excitation light source, a first filter, a first convex lens, a reflector, a dichroic mirror, a second convex lens, a third convex lens, a second filter and a detection device; the light source irradiated by the excitation light source passes through the first filter and the first convex lens in sequence and then directly hits the reflector, and then the light path direction is reflected on the dichroic mirror, and the dichroic mirror converts the light path direction and vertically shoots it to the target tube to be detected PCR reaction tube through the second convex lens; the fluorescence excited from the target tube to be detected PCR reaction tube passes through the second convex lens, the dichroic mirror, the third convex lens and the second filter in sequence and then enters the detection device for detection; The number of the optical detection channels is multiple; each fluorescence corresponds to at least two optical detection channels; the excitation light source is LED; the detection device is PD, APD, PMT or MPPC; The scanning motion assembly is used to install the optical reader and drive the optical reader to scan along the horizontal and vertical axes on a two-dimensional plane; the horizontal axis of the scanning motion assembly is guided by double guide rails, driven by a stepper motor and a synchronous belt; the vertical axis is guided by a single guide rail, driven by a brushless DC motor and a synchronous belt; the optical reader is fixed on the vertical axis; The PCR amplification temperature control component includes a base, a sealing structure, a thermal cycle module, a semiconductor refrigerator, a constant temperature water tank and a high-low tube adaptive structure; the base is used to install and fix the sealing structure, the constant temperature water tank, the semiconductor refrigerator and the constant temperature water tank; the top of the base is a mounting plate, and the middle of the mounting plate is provided with a mounting port, and the mounting port is used to install and fix the constant temperature water tank; anti-collision bars are respectively installed at the four corners of the base, and the height of the anti-collision bars is slightly higher than the height of the constant temperature water tank; the sealing structure is installed on the inner edge of the mounting port of the base; the thermal cycle module is used to carry PCR reaction tubes; the semiconductor refrigerator is installed at the bottom of the thermal cycle module; the semiconductor refrigerator is used to provide various temperatures required for PCR reactions in the PCR reaction tubes in the thermal cycle module; a constant temperature water tank is installed at the bottom of the semiconductor refrigerator; The thermal cover plate is provided with light holes corresponding to the bottom thermal cycle modules one by one; a lens is provided in the light holes; the sealing structure of the thermal cover and the PCR amplification temperature control component can form a sealed space; the sealing structure is made of a flexible and high temperature resistant material; a heating device is installed on the thermal cover for auxiliary heating; the sealing structure and the thermal cover are fitted in time to achieve a sealing effect; The lens in the light-through hole is a convex lens; the optical reader receives the fluorescence excited by the system in the PCR reaction tube on the lower constant temperature water tank through the convex lens in the light-through hole of the heat cover; the heat cover is located directly above the PCR amplification temperature control component; sliders are installed on both sides of the heat cover and can be extended and retracted; The lifting motion component is a scissor-type lifting platform, and the lifting and lowering of the scissor-type lifting platform is controlled by a lifting motor; the lifting motion component drives the PCR amplification temperature control component to rise and fall; when the PCR reaction tube needs to be placed or taken out, the lifting motion component vertically lifts the PCR amplification temperature control component to the upper part of the sampling port, which is convenient for taking and placing the PCR reaction tube, and also prevents the PCR reaction tube from falling into the bin; The high-low tube adaptive structure is used to adapt to the different heights of PCR reaction tubes in the constant temperature water tank; the high-low tube adaptive structure consists of a support plate and a lifting plate; the support plate is installed at the bottom of the PCR amplification temperature control component; guide columns are provided at the four corners of the support plate; springs are installed outside the guide columns; the lifting plate is installed on the top of the scissor-type lifting platform; The constant temperature water tank is used for heat dissipation of semiconductor refrigerators; the constant temperature water tank dissipates heat through the flow of refrigerant in the heat dissipation block; a double "bow"-shaped channel is arranged inside the heat dissipation block; the double "bow"-shaped channel is composed of two "bow"-shaped channels in opposite directions interlaced with each other; the double "bow"-shaped channel is provided with a liquid inlet and a liquid outlet, both of which are located on the same side of the bottom of the constant temperature water tank; the liquid inlet is connected to the water pump through a pipeline; the liquid outlet is connected to the radiator inlet through a pipeline; The liquid cooling system is used for cooling the refrigerant, and the liquid cooling system consists of a water pump, a liquid storage tank, a pipeline and a radiator; the liquid storage tank, the radiator and a number of partitions form an independent chamber, the bottom of the chamber is a rear air inlet, and one side is a heat dissipation port, which is isolated from other structures inside the instrument; the water pump is used to extract the refrigerant in the liquid storage tank and input it into the constant temperature water tank through a pipeline; the water pump outlet is connected to the liquid inlet of the constant temperature water tank through a pipeline; the water pump liquid extraction port is connected to the liquid storage tank outlet; the liquid storage tank is used to store refrigerant; the liquid storage tank inlet is connected to the heat pipe outlet of the radiator; the radiator is used to dissipate heat for the refluxed refrigerant, and the radiator has a built-in radiator-shaped heat pipe; the inlet of the heat pipe is connected to the liquid outlet of the constant temperature water tank through a pipeline, which is used to cool the refluxed refrigerant; a number of heat sinks are provided on the heat pipe body; a heat dissipation fan is also installed on the radiator to dissipate heat for the heat pipe.

2. A high-efficiency fluorescence quantitative PCR instrument according to claim 1, characterized in that: The flexible high temperature resistant material is a silicone material.

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