WO2017121238A1

WO2017121238A1 - Thermal cycle reaction component and real-time detection device having same

Info

Publication number: WO2017121238A1
Application number: PCT/CN2016/112367
Authority: WO
Inventors: 邱海维; 虞之龙; 胡琬璐
Original assignee: 北京酷搏科技有限公司
Priority date: 2016-01-15
Filing date: 2016-12-27
Publication date: 2017-07-20
Also published as: CN106978328A

Abstract

Provided are a thermal cycle reaction component (100) and a real-time detection device having same, wherein the thermal cycle reaction component (100) comprises a plurality of cylindrical heating components (10), wherein the plurality of cylindrical heating components (10) are arranged at intervals, and at least two cylindrical heating components (10) have different heating temperatures; and capillary prefabricated components (20), wherein the capillary prefabricated components (20) are arranged on the plurality of cylindrical heating components (10), solving the problem in the prior art that it is inconvenient to change the capillary.

Description

Thermal cycle reaction component and real-time detecting device therewith

Technical field

The invention relates to the field of polymerase chain reaction technology, in particular to a thermal cycle reaction component and a real-time detecting device therewith.

Background technique

Polymerase Chain Reaction (PCR) is a molecular biology technique used to amplify and amplify specific DNA fragments. With this PCR technique, a large amount of DNA can be generated by thermal cycling from a very small amount of DNA in vitro. Real-time Quantitative PCR Detecting System (QPCR) is a method in which the total amount of products after each thermal cycle is measured by adding a fluorescent chemical substance to a common PCR system, and the result can be obtained quickly. The data is relatively stable. The QPCR device includes a thermal cycler for the PCR reaction and a fluorometer for detecting the fluorescence of the reaction product. The thermal cycler consists of a thermal block, a temperature control circuit and a heat sink. Among them, the thermal block is used to conduct heat to the reaction tube containing the sample. The fluorometer includes a light source that emits excitation light, an optical component for selecting and conducting a fluorescent signal, and a light sensor that measures the intensity of the fluorescent signal. In the QPCR device, after each thermal cycle is completed, the fluorescence value of the sample is measured by the light sensor under excitation of the light source, thereby showing the real-time progress of the PCR. By detecting the fluorescence intensity of the sample in real time, the initial concentration of a particular DNA sequence in the sample to be tested can be accurately reversed.

As shown in FIG. 1, the prior art thermal cycler specifically includes an annular heating block 1 and a capillary tube 2 having a hollow portion in the middle, and the inside of the heating block 1 is divided into two layers so that The temperature outside the heating block 1 is different from the temperature inside the heating block 1, and the capillary 2 passes through the hollow portion of the heating block 1 and is wound around the heating block 1, and is wound a plurality of turns to conduct different temperatures to the capillary light, thereby enabling It is subjected to thermal cycle heating.

In the prior art, when the thermal cycler is used for the PCR reaction, the residue of the former sample in the capillary 2 may contaminate the new sample, resulting in inaccurate reaction. Therefore, the capillary needs to be replaced before the new sample is detected. In the prior art, when the capillary 2 is replaced, it is necessary to sequentially take out each of the capillaries 2 and re-wrap the new capillaries 2, making it extremely inconvenient to replace the capillaries 2.

Summary of the invention

The present invention provides a thermal cycle reaction assembly and a real-time detection device therewith to solve the problem of inconvenience in replacing the capillary tube in the prior art.

According to an aspect of the invention, there is provided a thermal cycle reaction assembly comprising a plurality of cylindrical heating assemblies, a plurality of cylindrical heating assemblies spaced apart, at least two cylindrical heating assemblies having different heating temperatures; The prefabricated assembly, the capillary prefabricated assembly is disposed on a plurality of cylindrical heating assemblies.

Further, the capillary prefabrication assembly comprises a first stage capillary tube, the first stage capillary tube is used for winding on the plurality of column heating assemblies; the second stage capillary tube is used for winding on one of the column heating elements, first Segment capillary The second segment of the capillary is in communication with each other, and the first segment of the capillary tube and the second segment of the capillary tube are respectively wound on the plurality of columnar heating assemblies with a plurality of layers, and the adjacent two layers of the capillary tubes are fixed to each other.

Further, the cylindrical heating assembly includes a housing, the capillary prefabricated assembly is wound on the outer wall of the housing; the heating portion is disposed inside the housing; the temperature controller is electrically connected to the heating portion, and the temperature controller is used to control the cylindrical heating assembly temperature.

Further, the thermal cycle reaction assembly further includes a propulsion mechanism disposed at the inlet end of the capillary prefabrication assembly.

Further, the thermal cycle reaction assembly further includes a collection device disposed at the outlet end of the capillary preform assembly.

Further, the thermal cycle reaction assembly further includes a mount assembly, and the plurality of post heating assemblies are fixedly disposed on the mount assembly.

Further, the fixing base assembly includes a fixing seat having a plurality of spaced apart first fixing holes, one ends of the plurality of cylindrical heating assemblies are disposed in the plurality of first fixing holes one by one; the limiting seat has a plurality of The second fixing holes are spaced apart from each other, and the spacing distance of the second fixing holes is greater than the spacing distance of the first fixing holes, and the other ends of the plurality of cylindrical heating assemblies are disposed in the plurality of second fixing holes one by one.

Further, the cylindrical heating assembly includes a first cylindrical heating assembly and a second cylindrical heating assembly, the first cylindrical heating assembly and the second cylindrical heating assembly are spaced apart, the heating temperature of the first cylindrical heating assembly is second The heating temperature of the cylindrical heating assembly is different, and the capillary prefabrication assembly is disposed on the first cylindrical heating assembly and the second cylindrical heating assembly.

According to another aspect of the present invention, there is provided a real-time detecting device comprising a thermal cycle reaction assembly, the thermal cycle reaction assembly being the thermal cycle reaction assembly provided above, and a light-emitting portion for emitting excitation light to the capillary prefabricated assembly a fluorescence detecting component for detecting fluorescence generated by irradiation of the sample in the capillary prefabricated component through the light emitting portion.

Further, the light emitting portion includes a light source for emitting excitation light to the capillary prefabrication assembly; an excitation band pass filter disposed on the optical path of the excitation light, and an excitation band pass filter for adjusting the wavelength of the excitation light.

Further, the fluorescence detecting component comprises a fluorescent converging lens for concentrating the fluorescence generated by the sample in the capillary prefabricated component; the detecting sensor is disposed on the optical path of the fluorescent condensed lens condensing; the band pass filter is disposed on the fluorescent concentrating lens and detecting Between the sensors, a bandpass filter is used for wavelength selection, and a detection sensor is used to detect fluorescence after wavelength selection by the bandpass filter.

Further, the fluorescence detecting component comprises a fluorescent converging lens for concentrating the fluorescence generated by the sample in the capillary prefabrication component; the detecting sensor is disposed on the optical path of the fluorescent light concentrated by the fluorescent converging lens; and the plurality of sets of band pass filters are switchably disposed at Between the fluorescent concentrating lens and the detecting sensor, the wavelength of the fluorescence condensed by the condensing lens is selected by any one of the band pass filters, and the detecting sensor is used for detecting the fluorescence after the wavelength selection of the band pass filter.

Further, the fluorescence detecting component further comprises a bracket, and the plurality of sets of band pass filters are disposed on the bracket; the motor and the motor are drivingly connected with the bracket, and the motor drives the bracket to rotate to switch the plurality of sets of band pass filters.

By applying the technical solution of the present invention, the capillary prefabricated assembly is disposed on the plurality of cylindrical heating assemblies, and when the capillary prefabricated assembly is replaced, the capillary prefabricated assembly is entirely removed from the cylindrical heating assembly, and the new capillary is removed. Pre The whole assembly of the assembly is arranged on the cylindrical heating assembly to complete the replacement, so that the worker can replace the capillary prefabricated assembly and improve the detection efficiency.

DRAWINGS

The accompanying drawings, which are incorporated in the claims of the claims In the drawing:

1 is a schematic structural view of a thermal cycler provided by the prior art;

2 is a schematic structural view of a thermal cycle reaction assembly provided in Embodiment 1;

Figure 3 is a schematic view showing the structure of the capillary prefabrication assembly of Figure 2;

4 is a schematic structural view of a capillary prefabrication assembly provided in Embodiment 2;

Figure 5 shows a cross-sectional view of the cylindrical heating assembly of Figure 2 and the base;

Figure 6 is a schematic view showing the temperature distribution of the cylindrical heating assembly for heating the capillary prefabricated assembly;

FIG. 7 is a schematic structural diagram of a real-time detecting apparatus provided in Embodiment 3;

FIG. 8 is a schematic structural diagram of a real-time detecting apparatus provided in Embodiment 4.

Wherein, the above figures include the following reference numerals:

1, heating block; 2, capillary; 10, cylindrical heating assembly; 11, housing; 12, heating portion; 13, temperature controller; 20, capillary prefabricated assembly; 21, first segment capillary; Capillary; 30, propulsion mechanism; 40, collecting device; 51, fixing seat; 52, limiting seat; 100, thermal cycle reaction component; 60, light-emitting part; 61, light source; 62, excitation band-pass filter; Detection component; 71, fluorescent convergence lens; 72, detection sensor; 73, band pass filter; 74, bracket; 75, motor.

detailed description

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

As shown in FIGS. 2 to 4, the first embodiment is a thermal cycle reaction assembly including a plurality of cylindrical heating assemblies 10 and a capillary prefabrication assembly 20. Therein, a plurality of cylindrical heating assemblies 10 are spaced apart, and the heating temperatures of the cylindrical heating assemblies 10 are different. Capillary preform assembly 20 is disposed on a plurality of cylindrical heating assemblies 10. The number and temperature of the cylindrical heating assembly 10 can be specifically set according to the test. In the PCR reaction, the temperature in the denaturation stage is between 92 ° C and 98 ° C, and in the primer binding and DNA extension stages, the required temperature is Between 50 ° C and 75 ° C. Therefore, in the present embodiment, two cylindrical heating assemblies 10 are provided, wherein the temperature of one cylindrical heating assembly 10 is set at 95 ° C, and the temperature of the other cylindrical heating assembly 10 is set at 60 ° C, in order to facilitate the solution. It is stated that the temperature of the large-diameter cylindrical heating assembly 10 is set at 60 ° C, and the temperature of the small-diameter cylindrical heating assembly 10 is set at 95 ° C. Through a small diameter column The heating assembly 10 denatures the sample and anneals and extends the sample through the large diameter cylindrical heating assembly 10. Figure 6 shows the temperature distribution of the sample in the capillary preform assembly 20 during the actual heating process. The temperature at a is 60 ° C, the temperature at b is 60 ° C to 95 ° C, the temperature at c is 95 ° C, and the temperature at d is 95. °C to 60 °C.

With the technical solution of the present invention, the capillary prefabrication assembly 20 is disposed on the plurality of cylindrical heating assemblies 10, and the capillary prefabricated assembly 20 can be completely removed from the cylindrical heating assembly 10 when the capillary prefabricated assembly 20 is replaced. Then, the new capillary prefabrication assembly 20 is integrally disposed on the cylindrical heating assembly 10 to complete the replacement, so that the worker can replace the capillary prefabrication assembly 20, thereby improving the detection efficiency. Moreover, the winding mode facilitates forming the capillary preform assembly 20 into an integral annular structure such that when the capillary preform assembly 20 is repositioned on the cylindrical heating assembly 10, the capillary preform assembly 20 is integrally nested on the cylindrical heating assembly 10. Just go up.

Specifically, after the capillary prefabrication assembly 20 is wound around the two cylindrical heating assemblies 10, the capillary prefabricated assembly 20 can be bonded into a unitary structure by an adhesive, or the outer wall of the capillary can be melted or softened by heating to adhere to each other. Hehe. The capillary prefabricated assembly 20 can be turned into an integral annular structure, which facilitates overall removal or installation, thereby further saving installation time and improving installation efficiency.

In the present embodiment, the cylindrical heating assembly 10 is provided in a cylindrical shape to facilitate the mounting of the capillary prefabricated assembly 20 and is easy to manufacture. The cylindrical heating assembly 10 may be provided as other cylindrical structures such as a prismatic shape and an elliptical cross section as needed.

Optionally, the diameters of the two cylindrical heating assemblies 10 are 10 mm and 20 mm, respectively, and the center distance between the two cylindrical heating assemblies 10 is 40 mm, but the diameters and spacings of the two cylindrical heating assemblies 10 are not limited thereto, and the sizes and intervals thereof. In relation to the time required for the different PCR reactions, the diameter of the cylindrical heating assembly 10 is typically set between 2 mm and 50 mm. And because the number of heating cycles required for each PCR reaction is different, the number of turns of the capillary preform assembly 20 wound on the cylindrical heating assembly 10 is also different. Therefore, the length of the cylindrical heating assembly 10 is set to be between 20 mm and 200 mm.

3 and 4 illustrate the structure of two different capillary preform assemblies 20, except that the capillary preform assembly 20 of FIG. 3 includes a first length of capillary 21 and a second length of capillary 22. Wherein, the first stage capillary 21 sequentially bypasses the plurality of cylindrical heating assemblies 10 and is wound around the plurality of cylindrical heating assemblies 10, and the second length of the capillary tubes 22 is wound around one of the cylindrical heating assemblies 10. Further, the first-stage capillary tube 21 and the second-stage capillary tube 22 are wound on the plurality of column-shaped heating assemblies 10, respectively, and the adjacent upper and lower layers of the capillary tubes are fixed to each other by the above-described bonding method.

In the present embodiment, the second stage capillary 22 is wound around the small diameter cylindrical heating assembly 10. Since in the heat-activated DNA polymerase, an antibody, an inhibitor or other substance capable of blocking the activity of the polymerase at a normal temperature is usually added. Therefore, before the start of the PCR reaction, the inhibition of the DNA polymerase is usually released by heating, so that the PCR reaction can proceed normally. Here, in order to obtain a sufficient hot start temperature, in the present embodiment, the second stage capillary 22 is wound 10 times on the small diameter cylindrical heating assembly 10. And the heating temperature is generally at 95 ° C, and the passage time is from 1 to 10 minutes. By the structure of the capillary prefabrication assembly 20 shown in Fig. 3, the inhibition of the DNA polymerase can be released by winding the second length of the capillary 22 on the columnar heating assembly 10 having a higher temperature without separate heating means. Heating the sample facilitates the progress of the PCR reaction and reduces the number of reaction devices required.

Specifically, the material of the capillary prefabrication assembly 20 is polytetrafluoroethylene, but not limited to polytetrafluoroethylene, and may be polyvinylidene fluoride, perfluoroethylene propylene polymer, polypropylene, polycarbonate, polydimethylsiloxane. Alkane, fused silica, silicate glass, borate glass or composites, copolymers thereof, and the like. It is also possible that the inner wall of the capillary made of one of the above materials is covered with another material or more. In the present embodiment, the capillary preform assembly 20 has an outer diameter of 0.6 mm and an inner diameter of 0.3 mm, but is not limited thereto. Depending on the flow rate of the PCR reaction, the amount of sample, and the rate of thermal conductivity, the outer diameter of the capillary preform assembly 20 can be set between 0.2 mm and 2.0 mm, and the inner diameter of the capillary preform assembly 20 can be set between 0.05 mm and 1.8 mm.

As shown in FIG. 5, a cross-sectional view of a cylindrical heating assembly 10 is provided, wherein the cylindrical heating assembly 10 includes a housing 11, a heating portion 12, and a temperature controller 13. The capillary prefabrication assembly 20 is wound around the outer wall of the housing 11, the heating portion 12 is disposed inside the housing 11, and the temperature controller 13 is electrically connected to the heating portion 12, and the heating of the cylindrical heating assembly 10 can be controlled by the temperature controller 13. temperature. Optionally, the heating portion 12 may be a heating core, and may be composed of a resistance wire, or may be provided with a heating sheet, an electric heating tube, a heating wire, a circuit board, a Peltier unit, or the like. The heating portion 12 may heat the casing 11 by generating heat radiation by electric current, or may heat the casing 11 by means of heat convection, such as heating water or other liquid, gas, or phase change heat conduction. Further, the heat generated by the heating unit 12 is detected and adjusted in real time by the temperature controller 13, so that the heat supplied to the capillary prefabrication unit 20 can be accurately controlled to ensure the normal progress of the PCR reaction.

Wherein, the thermal cycle reaction assembly further includes a propulsion mechanism 30 disposed at the inlet end of the capillary prefabrication assembly 20, and the sample liquid is pushed by the propulsion mechanism 30 to move within the capillary prefabrication assembly 20. In the present embodiment, the advancement mechanism 30 is disposed at the port of the second stage capillary 22, and the advancement mechanism 30 can be configured as a syringe pump. However, the present invention is not limited thereto, and the propulsion mechanism 30 may be a peristaltic pump, an air pressure pump, or a liquid generated by compressing a gas or an electrolyte body to generate a gas, or a method of using a gas to heat volume expansion to realize a sample liquid in a capillary prefabricated assembly. One-way or two-way movement in 20. In the present embodiment, the pushing flow rate of the propulsion mechanism 30 is about 0.4 uL/s, and after 600 s, it can be filled to the 36th turn of the first stage capillary 21 to complete the PCR reaction. However, the pushing speed and the advancing time of the reaction sample are not limited to this. According to the inner diameter of the capillary prefabrication assembly 20 and the number of thermal cycles for the PCR reaction, the pushing flow rate of the propulsion mechanism 30 can vary from 0.01 uL/s to 10 uL/s, and the advancement time of a single sample to be tested is usually between 300 s and 1200 s. between.

In the first embodiment of the present invention, the thermal cycle reaction assembly further includes a collection device 40 disposed at the outlet end of the capillary prefabrication assembly 20. In the present embodiment, the collecting device 40 is disposed at the port of the first stage capillary 21, and the reaction liquid after the reaction is collected by the collecting device 40. Wherein, the collecting device 40 can be arranged as a flexible balloon, or can be set in other containers of constant or variable volume, such as a syringe, a plastic bottle or the like. The collecting device 40 may be filled with air, a water absorbing material (such as a sponge), a substance reactive with water (such as quick-drying agent such as quicklime, gypsum, cobalt chloride), a DNA adsorbing material (such as porous silicon oxide), or the like.

In the first embodiment of the present invention, the thermal cycle reaction assembly further includes a fixing base assembly, wherein the plurality of cylindrical heating assemblies 10 are fixedly disposed on the fixing base assembly. To avoid sloshing of the cylindrical heating assembly 10 during the test, the test was carried out.

Optionally, the fixing base assembly includes a fixing base 51 and a limiting seat 52. The fixing base 51 has a plurality of spaced apart first fixing holes, and one ends of the plurality of cylindrical heating assemblies 10 are disposed one by one. In a plurality of first fixing holes. A plurality of second fixing holes are also disposed on the limiting seat 52, and the other ends of the plurality of cylindrical heating assemblies 10 are disposed in the plurality of second fixing holes one by one. Specifically, the spacing distance of the second fixing holes is greater than the spacing distance of the first fixing holes, so as to increase the column shape There is a pre-tightening force between the heat assembly and the two bases, so that the plurality of cylindrical heating assemblies 10 can be fastened to the fixing base 51 and the limiting seat 52. When the thermal cycle reaction assembly is installed, one end of the plurality of cylindrical heating assemblies 10 is first fixed on the fixing base 51, and the capillary prefabricated assembly 20 is placed on the plurality of cylindrical heating assemblies 10, and then the limiting seat 52 is placed. Fixedly disposed at the other end of the plurality of cylindrical heating assemblies 10, since the spacing distance of the second fixing holes is greater than the spacing distance of the first fixing holes, the plurality of cylindrical heating assemblies 10 are moved to both sides, thereby being able to open the capillary prefabrication The assembly 20 secures the capillary preform assembly 20 to the plurality of cylindrical heating assemblies 10.

When the PCR reaction is carried out through the thermal cycle reaction module, if the sample volume is small, the capillary prefabrication assembly 20 may not be filled. At this point, another inert liquid that is immiscible with the sample can be selected as the medium carried by the sample to assist in pushing the sample forward in the capillary. In the present embodiment, silicone oil may be used as a medium for carrying the sample, but it is not limited to the use of silicone oil, and may be a fluoroalkane, a fluoroether, a liquid paraffin or the like. In this embodiment, a sample of 1 uL is mixed with 400 uL of silicone oil, and the sample is passed through the carrier medium, thus ensuring the successful completion of the PCR reaction.

As shown in FIG. 7, a third embodiment of the present invention is a real-time detecting device, which includes a thermal cycle reaction component 100, a light emitting portion 60, and a fluorescence detecting component 70. The thermal cycle reaction assembly 100 is the thermal cycle reaction assembly 100 provided by the above embodiment. The light emitting portion 60 is configured to emit excitation light to the capillary prefabrication assembly 20, and the fluorescence detecting member 70 is configured to detect the sample in the capillary prefabrication assembly 20 via the light emitting portion 60. The fluorescence produced by the irradiation.

Through the real-time detecting device, the samples in each layer of the capillary prefabrication assembly 20 can be simultaneously detected, so that the worker can obtain the detection data. And the device has a simple structure and is easy to operate. Compared with the conventional thermal cycler shown in FIG. 1, when detecting the same, it is necessary to arrange the light-emitting portion and a part of the fluorescent detecting member in the hollow portion, so that the hollow portion of the thermal cycler must satisfy the light-emitting portion and the fluorescence detection. The size of the components does not allow the thermal cycler to move toward miniaturization. In the real-time detecting device provided in the third embodiment, the light-emitting portion 60 and the fluorescence detecting member 70 need only be disposed on one side of the thermal cycle reaction assembly 100, and the size of the thermal cycle reaction assembly 100 may be set according to the PCR reaction. This simplifies the design of the thermal cycle reaction assembly 100.

Specifically, the light emitting portion 60 includes a light source 61 and an excitation band pass filter 62. The excitation light is emitted from the light source 61 to the capillary prefabrication unit 20, the excitation band pass filter 62 is disposed on the optical path of the excitation light, and the wavelength of the excitation light is adjusted by the excitation band pass filter 62. Alternatively, the light source 61 may be a white light source such as a halogen lamp or a xenon lamp, or may be a monochromatic or narrow wavelength light source such as an LED lamp or a laser.

Specifically, the fluorescence detecting section 70 includes a fluorescence condensing lens 71, a detecting sensor 72, and a band pass filter 73. The fluorescent concentrating lens 71 is used to condense the fluorescence generated by the sample in the capillary prefabrication assembly 20, and the detecting sensor 72 is disposed on the optical path of the fluorescence condensed by the condensing condenser lens 71. The band pass filter 73 is disposed on the fluorescent condensing lens 71 and the detecting sensor 72. Between, the band pass filter 73 is used for filtering, and the detecting sensor 72 is for detecting the fluorescence filtered by the band pass filter 73. By detecting the sensor 72, the fluorescence intensity of the sample passing through the capillary can be obtained in real time, and the initial concentration of the specific DNA sequence can be obtained by the fluorescence intensity.

Alternatively, the fluorescence concentrating lens 71 may be provided as an aspherical lens or an aspherical lens group, and the detecting sensor 72 may be provided as an area CMOS sensor or an area CCD sensor. The band pass filter 73 may be an absorptive filter or a reflective filter.

As shown in FIG. 8, the fourth embodiment provides a real-time detecting device, which differs from the third embodiment only in that the fluorescent detecting component 70 includes a plurality of sets of band pass filters 73 and a plurality of sets of band pass filters. 73 is switchably disposed between the fluorescence condensing lens 71 and the detecting sensor 72, and filters the fluorescence condensed by the condensing lens 71 by any one of the band pass filters 73. The detecting sensor 72 is configured to detect the filtered by the band pass filter 73. Fluorescence. Thus, different bandpass filters 73 can be adjusted to obtain fluorescence of different wavelength bands, and fluorescence of different wavelength bands can be detected.

Specifically, the fluorescence detecting component 70 further includes a bracket 74 and a motor 75, wherein the plurality of sets of band pass filters 73 are disposed on the bracket 74, the motor 75 is drivingly coupled to the bracket 74, and the motor 75 drives the bracket 74 to rotate when needed. The plurality of sets of band pass filters 73 are switched.

By the real-time detecting device provided by the invention, the reaction time of the existing QPCR can be shortened from 1 to 2 hours to 5 to 20 minutes, which can greatly shorten the reaction time and increase the analysis speed. And the device has a simple structure and a small overall size, which is convenient for the staff to use.

The above description is only the preferred embodiment of the present invention, and is not intended to limit the present invention, and various modifications and changes can be made to the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and scope of the present invention are intended to be included within the scope of the present invention.

Claims

A thermal cycle reaction assembly characterized in that the thermal cycle reaction assembly comprises:

a plurality of cylindrical heating assemblies (10), a plurality of said cylindrical heating assemblies (10) are spaced apart, and at least two of said cylindrical heating assemblies (10) have different heating temperatures;

A capillary prefabricated assembly (20) is disposed on a plurality of the cylindrical heating assemblies (10).
The thermal cycle reaction assembly of claim 1 wherein said capillary preform assembly (20) comprises:

a first section of the capillary (21), the first section of the capillary (21) for winding around the plurality of cylindrical heating elements (10);

a second stage capillary tube (22) for winding on one of the columnar heating assemblies (10), the first stage capillary tube (21) and the second stage capillary tube (22) communicating with each other, the first segment capillary tube (21) and the second segment capillary tube (22) are respectively wound with a plurality of layers on the plurality of columnar heating assemblies (10), adjacent two layers of capillaries They are fixed together.
The thermal cycle reaction assembly of claim 1 wherein said cylindrical heating assembly (10) comprises:

a casing (11), the capillary prefabricated assembly (20) is wound on an outer wall of the casing (11);

a heating portion (12) disposed inside the casing (11);

A temperature controller (13) is electrically connected to the heating portion (12) for controlling the temperature of the cylindrical heating assembly (10).
The thermal cycle reaction assembly of claim 1 wherein said thermal cycle reaction assembly further comprises:

A propulsion mechanism (30) is disposed at the inlet end of the capillary preform assembly (20).
The thermal cycle reaction assembly of claim 1 wherein said thermal cycle reaction assembly further comprises:

A collection device (40) is disposed at the outlet end of the capillary preform assembly (20).
The thermal cycle reaction assembly of claim 1 wherein said thermal cycle reaction assembly further comprises:

A fixing base assembly, a plurality of the cylindrical heating assemblies (10) are fixedly disposed on the fixing base assembly.
The thermal cycle reaction assembly of claim 6 wherein said mount assembly comprises:

a fixing base (51) having a plurality of spaced apart first fixing holes, one end of each of the plurality of cylindrical heating assemblies (10) being disposed in a plurality of the first fixing holes in one-to-one correspondence;

a limiting seat (52) having a plurality of spaced apart second fixing holes, wherein the second fixing holes are spaced apart by a distance greater than a spacing distance of the first fixing holes, and the plurality of cylindrical heating assemblies (10) The other ends are disposed one by one in a plurality of the second fixing holes.
The thermal cycle reaction assembly of claim 1 wherein said cylindrical heating assembly (10) comprises a first cylindrical heating assembly and a second cylindrical heating assembly, said first cylindrical heating assembly and The second cylindrical heating assembly is spaced apart, the heating temperature of the first cylindrical heating assembly is different from the heating temperature of the second cylindrical heating assembly, and the capillary prefabricated assembly (20) is disposed on the first column And heating the assembly and the second cylindrical heating assembly.
A real-time detecting device, wherein the real-time detecting device comprises:

a thermal cycle reaction assembly (100), the thermal cycle reaction assembly (100) being the thermal cycle reaction assembly (100) according to any one of claims 1 to 8;

a light emitting portion (60) for emitting excitation light to the capillary prefabrication assembly (20);

A fluorescence detecting member (70) for detecting fluorescence generated by the sample in the capillary prefabrication assembly (20) being irradiated by the light emitting portion (60).
The real-time detecting device according to claim 9, wherein the light emitting portion (60) comprises:

a light source (61) for emitting excitation light to the capillary prefabrication assembly (20);

An excitation bandpass filter (62) is disposed on the optical path of the excitation light, and the excitation bandpass filter (62) is configured to adjust a wavelength of the excitation light.
The real-time detecting device according to claim 9, wherein the fluorescence detecting unit (70) comprises:

a fluorescent converging lens (71) for concentrating fluorescence generated by the sample in the capillary prefabricated assembly (20);

a detecting sensor (72) disposed on the optical path of the fluorescent light condensed by the fluorescent concentrating lens (71);

a band pass filter (73) disposed between the fluorescent converging lens (71) and the detecting sensor (72) for performing wavelength selection, the detecting sensor (72) ) for detecting fluorescence after wavelength selection by the band pass filter (73).
The real-time detecting device according to claim 9, wherein the fluorescence detecting unit (70) comprises:

a fluorescent converging lens (71) for concentrating fluorescence generated by the sample in the capillary prefabricated assembly (20);

a detecting sensor (72) disposed on the optical path of the fluorescent light condensed by the fluorescent concentrating lens (71);

A plurality of sets of band pass filters (73) are switchably disposed between the fluorescent concentrating lens (71) and the detecting sensor (72), and the fluorescent light is applied to the fluorescent light by any one of the band pass filters (73) The fluorescence concentrated by the converging lens (71) is used for wavelength selection, and the detecting sensor (72) is for detecting fluorescence selected by the wavelength of the band pass filter (73).
The real-time detecting device according to claim 12, wherein the fluorescence detecting unit (70) further comprises:

a bracket (74), a plurality of sets of the band pass filters (73) are disposed on the bracket (74);

a motor (75) drivingly coupled to the bracket (74), the motor (75) driving the motor The bracket (74) rotates to switch a plurality of sets of the band pass filters (73).