KR20220135929A - PCR apparatus for real-time controlling well temperature individually, temperature controlling method for the same PCR apparatus and sample detecting method for the same PCR apparatus - Google Patents

Abstract

The present invention relates to a PCR device for individually controlling a well temperature in real time, a method for controlling a temperature of a PCR device, and a method for detecting a sample of a PCR device. By including a heating device surrounded by a heating film for each well and facilitating individual heating of wells, a purpose of the present invention is to provide the PCR device for individually controlling a well temperature in real time and the method for controlling a temperature of a PCR device. Another purpose of the present invention is to provide the PCR device and the method for detecting a sample of a PCR device capable of optimizing detecting efficiency and accuracy by arranging a plurality of wells in a circle shape and rotating an optical system to irradiate and detect light.

Description

PCR apparatus for individually and real-time controlling well temperature, a method for controlling a temperature of the PRC apparatus, and a method for detecting a sample of the PCR apparatus {PCR apparatus for real-time controlling well temperature individually, temperature controlling method for the same PCR apparatus and sample detecting method for the same PCR apparatus}

The present invention relates to a PCR apparatus, and more particularly, a real-time PCR apparatus capable of individually controlling the temperature of each well in real time for a plurality of wells, a temperature control method of the PCR apparatus, and sample detection it's about how

PCR (polymerase chain reaction) is a method of amplifying a specific target genetic material to be detected. By repeatedly heating and cooling a sample containing a nucleic acid, a region having a specific nucleotide sequence of the nucleic acid is replicated in a chain. This is a technique for exponentially amplifying nucleic acids having a specific nucleotide sequence region. PCR may be performed in a series of thermoenzymatic reaction steps such as denaturation, annealing, and extension. Briefly, in the denaturation step, two strands of DNA are separated by heating, and each DNA serves as a template. In the binding step, primers are bound to the template DNA. At this time, the accuracy of the reaction can be improved by adjusting the temperature appropriately. In the elongation phase, heat-resistant DNA polymerase creates new DNA from the template DNA. If the polymerase chain reaction is performed once, the genetic material is amplified twice. Therefore, if the polymerase chain reaction is repeated n times, in theory, the gene of 2 to n times the gene can be amplified, exponentially by repeating the reaction. phosphorus amplification is possible.

A PCR device is a device made for the purpose of allowing a PCR reaction to occur more efficiently with respect to a large amount of sample. As can be intuitively seen from the above description, since temperature control is a key and important part in the PCR reaction, the PCR device is generally equipped with a heating device and a cooling device for controlling the temperature of the sample. In addition, in order to obtain a large amount of and, if necessary, a variety of results with one PCR device, a PCR device is generally provided with a plurality of wells, and a tube containing a sample in the plurality of wells is provided, respectively. In the accommodated state, the temperature control of the sample is performed using the heating device and the cooling device described above. The basic configuration of such a PCR device is well disclosed in Korean Patent Registration No. 2071058 ("A device and method for automated extraction and delivery of nucleic acids for gene amplification," 2020.01.21., hereinafter 'prior literature').

As well shown in the preceding documents, in general, in a PCR apparatus, a heating block using a Peltier element or the like is employed as a heating device, and a cooling fan is employed as a cooling device. In more detail, in general, a PCR device includes a well formed in the form of a circular through-hole on a plate, a plurality of arranged wells and a tube, which is inserted into the well and fixedly supported. In this configuration, a heating block is installed in the lower part of the tube and the temperature is controlled through heat transfer by contacting the tube. This method has the advantage of being able to control the temperature in a contact type (heat conduction method) in a fixed and non-moving device, so that the response speed is fast and it is very easy to measure the accuracy of the temperature.

However, in the case of the conventional PCR apparatus, since the temperature of a plurality of wells is controlled at once using one heating block, there is a disadvantage in that it is impossible to control the temperature of each well. Even if the heating block is divided into smaller units, as a result, individual temperature control is possible only in the block unit range, so it is difficult to realize complete individual well temperature control.

In addition, referring to the prior literature, one optical system for light irradiation and detection is provided per well, so that light irradiation and detection are performed in all tubes at once. However, in this way, the number of optical systems is excessively increased, and not only the manufacturing cost is excessively increased, but also there is a problem in that the distance between the optical systems cannot be sufficiently widened, so that the signal interference is greatly generated, thereby lowering the detection accuracy.

Therefore, in general, in many conventional PCR devices, unlike the prior literature, a smaller number of optical systems than the tube are provided and configured to perform light irradiation and detection while moving them. At this time, many conventional PCR devices use a method of arranging tubes in two dimensions and performing light irradiation and detection while advancing the optical system in the horizontal and vertical directions. However, in this case, the optical system must stop in order to irradiate or detect light from one tube, and the optical system must proceed to perform light irradiation and detection in the next tube. will do In this process, the optical system may be subjected to unnecessary shocks by inertia, and there is a risk of increased dynamic instability because the actuator of the moving device has to repeatedly stop and proceed.

1. Korean Patent Registration No. 2071058 ("A device and method for automated extraction and delivery of nucleic acids for gene amplification", 2020.01.21.)

Accordingly, the present invention has been devised to solve the problems of the prior art as described above, and an object of the present invention is to enable individual heating of wells by providing a heating device in a form in which a heating film is surrounded for each individual well. An object of the present invention is to provide a PCR apparatus for individually controlling the well temperature in real time and a method for controlling the temperature of the PCR apparatus. Another object of the present invention is to provide a PCR apparatus for optimizing detection efficiency and accuracy by arranging a plurality of wells in a circle and performing light irradiation and detection while rotating an optical system, and a sample detection method of the PCR apparatus.

The PCR apparatus 100 for individually controlling the well temperature of the present invention in real time to achieve the above object heats and cools the sample accommodated in the tube 200 to generate a polymerase chain reaction (PCR) reaction, A PCR device (100) for detecting a target material in the sample using fluorescence generated by irradiating excitation light to the sample, comprising: an upper body (110); a lower body 120 in which a plurality of the tubes 200 are arranged in a circle at an upper end; It is accommodated in the upper body 110, is radially spaced at equal intervals, and includes a plurality of optical systems 132 that individually perform light irradiation and detection, respectively, and light irradiation with respect to the sample accommodated in the tube 200. and an optical module 130 for performing detection; a motor 140 accommodated in the upper body 110 and configured to rotate the optical module 130 so that light irradiation and detection are performed on the sample accommodated in each of the tubes 200 at a predetermined cycle; a heater module 150 including a heater structure 155 accommodated in the lower body 120 and provided to surround each of the tubes 200 so as to be able to individually heat the tubes 200; a cooling fan (160) accommodated in the lower body (120) and provided at the center of a circular arrangement of a plurality of the tubes (200) to forcibly blow air around the tube (200) to cool it; may include

The heater structure 155 is formed in the form of a metal pipe to accommodate a tube receiving portion 155a into which the tube 200 is inserted, and is formed in a film shape to surround the side surface of the tube receiving portion 155a. It may include a film heater unit 155b for heating and a temperature detecting unit attached to the tube receiving unit 155a to measure the temperature of the tube 200 in a contact manner.

In addition, the heater module 150 may be formed such that a through hole is formed in the center so that the air forcedly blown by the cooling fan 160 passes through the central through hole of the heater module 150 .

In addition, in the PCR apparatus 100, when heating, the heater module 150 uniformly heats a plurality of the tubes 200 or drives the heater structure 155 individually to each tube 200. may be individually heated, and when cooling, the cooling fan 160 may be formed to uniformly cool the plurality of tubes 200 .

At this time, the temperature control method of the PCR apparatus of the present invention, in the temperature control method of the PCR apparatus 100, the heater module 150 heating the plurality of tubes 200 uniformly the temperature rising step; a temperature lowering step in which the cooling fan 160 forcibly blows air to uniformly cool the plurality of tubes 200; Temperature control in which the heater module 150 individually drives the heater structures 155 provided in each tube 200 to individually heat the temperature of each tube 200 to maintain a predetermined reference temperature step; may include.

The optical module 130 includes a fixed plate 131 formed in a circular shape, an arrangement provided on the fixed plate 132 consisting of a plurality of the optical systems 132 , and an arrangement provided on the upper surface of the arrangement to form the structure of the arrangement. A first optical control substrate 133 for controlling the light irradiation and detection operation of the optical system 132 while supporting the upper surface, is provided laminated on the upper surface of the first optical control substrate 133, and the optical system 132 is irradiated with light and a second optical control board 134 for controlling a link operation with the motor 140 according to the detection operation.

In addition, the optical module 130 may be formed such that a hole is formed in the center so that the motor 140 is disposed in the center hole of the optical module 130 .

In addition, in the PCR apparatus 100, the optical system 132 emits light and detects the sample accommodated in the tube 200 while the motor 140 rotates the optical module 130 at a predetermined rotation speed. It can be formed to perform. In this case, the PCR apparatus 100 may adjust the rotation speed of the motor 140 according to a desired signal sampling rate.

At this time, the sample detection method of the PCR apparatus of the present invention comprises: a module rotation step in which the motor 140 rotates the optical module 130 at a predetermined rotation speed; a light irradiation step in which the optical system 132 irradiates excitation light to the sample accommodated in the tube 200; a photodetection step in which the optical system 132 detects a target material in the sample using the fluorescence generated from the sample accommodated in the tube 200; may include.

On the other hand, the PCR device 100, the upper body 110 and the lower body 120 may be formed to be able to open and close.

The upper body 110 is coupled to the upper body cover 111 forming a space therein, the lower end of the upper body cover 111, and the upper and lower cover through hole 112h at a position corresponding to the tube 200. The upper and lower cover 112, which is formed, is provided on the upper side of the upper and lower cover 112, the motor 140 is fixed in the center of the upper surface, and the fixing bracket through hole 113h is provided at a position corresponding to the tube 200. The formed fixing bracket 113, the optical module 130 and the rotation bracket 114 for coupling the rotation shaft of the motor 140, and a slip ring 115 for supplying power to the motor 140. can

In addition, the upper body 110 is provided between the fixing bracket 113 and the upper and lower cover 112 to uniformly heat the plurality of tubes 200 and a hot top to a position corresponding to the tube 200 . A hot top heater substrate 116 in which the heater substrate through hole 116h is formed may be further included.

In addition, the lower body 120, a plurality of circularly arranged lower upper cover through-holes 121h are formed, the tube 200 is inserted, and the lower upper cover 121 disposed at the lower end of the upper body 120. , the lower upper cover 121 is coupled to the upper end, the lower side cover 122 forming a space therein, is coupled to the lower end of the lower side cover 122 and the cooling fan 160 is fixed to the center of the upper surface The lower lower cover 123, which is provided on the lower side of the lower upper cover 121, is coupled to the upper surface of the heater module 140 to fix the heater module 140, and the heater is fixed at a position corresponding to the tube 200 A substrate through hole 124h is formed so that the heater structure 155 penetrates through the heater fixed substrate 124, and is formed in the center of the lower lower cover 123 and forcibly blows air by the cooling fan 160. A plurality of lower surface vents 125 to circulate with the outside, a plurality of side vents 126 formed on the upper side of the side of the lower side cover 122 to distribute the air forcedly blown by the cooling fan 160 with the outside. can do.

According to the present invention, since individual heaters are provided for each of the plurality of wells, there is a great effect of individually controlling the well temperature in real time. More specifically, in the present invention, individual heating is performed by providing one heater per well, and uniform cooling is performed for a plurality of wells using a cooling fan. The temperature of individual wells can be performed independently of each other and seamlessly in real time. Accordingly, there is a great effect that the temperature of each well can be more precisely and precisely adjusted to the desired reaction temperature. In addition, according to the present invention, since the well temperature can be individually controlled, there is also an effect that various applications are possible, such as reacting a variety of samples at once.

Meanwhile, according to the present invention, by providing an optical module capable of more efficiently detecting the fluorescence signal generated in a plurality of wells, there is also an effect of greatly improving the fluorescence signal detection efficiency and accuracy. More specifically, in the present invention, the wells are arranged in a circle, and the process of irradiating excitation light to a sample in a tube provided in a plurality of wells and detecting fluorescence with an optical module is performed together with a rotation operation. Accordingly, the detection of the fluorescence signal in the plurality of wells can be performed more efficiently, and the detection accuracy can be improved by adjusting the rotation speed of the optical module.

1 is an external perspective view when the PCR device of the present invention is closed.
Figure 2 is an external perspective view when the PCR device of the present invention is opened.
Figure 3 is an external side view and a cross-sectional side view of the PCR device of the present invention.
Figure 4 is an exploded perspective view of the upper body of the PCR device of the present invention.
Figure 5 is an exploded perspective view of the lower body of the PCR device of the present invention.
Figure 6 is an exploded perspective view of the optical module of the PCR device of the present invention.
7 is an exploded perspective view of the heater structure of the PCR device of the present invention.

Hereinafter, a PCR apparatus for individually controlling the well temperature in real time according to the present invention having the configuration as described above, a temperature control method of the PCR apparatus, and a sample detection method will be described in detail with reference to the accompanying drawings.

1 and 2 are external perspective views when the PCR device of the present invention is closed and opened, and Figure 3 is an external side view and a cross-sectional side view of the PCR device of the present invention. The PCR apparatus 100 of the present invention basically generates a PCR (polymerase chain reaction) reaction by heating and cooling the sample accommodated in the tube 200, and uses the fluorescence generated by irradiating the sample with excitation light. Detect my target substance. At this time, as shown in FIGS. 1 to 3 , the PCR device 100 of the present invention includes an upper body 110 and a lower body 120 , and an optical module 130 and an optical module 130 on the upper body 110 . A motor 140 is accommodated to perform light irradiation and detection, and a plurality of the tubes 200 are arranged in a circle at the upper end of the lower body 120 , and the heater module 150 is provided in the lower body 120 . And a cooling fan 160 is accommodated to perform heating and cooling.

At this time, in the present invention, individual temperature control is possible by providing a heating device that can be individually heated to each of the tubes 200 , and the optical module 130 is irradiated with light while rotating by the motor 140 . and to improve the efficiency and accuracy of sample detection by performing the detection. Hereinafter, each part will be described in more detail.

The upper body 110 is provided to accommodate the optical module 130 and the motor 140 therein as described above. 4 is an exploded perspective view of the upper body of the PCR apparatus of the present invention, and the detailed configuration of the upper body 110 will be described with reference to the cross-sectional view of FIG. 3 and the exploded perspective view of FIG.

The upper body 110 forms an inner space by combining the upper body cover 111 that forms a space therein and the upper lower cover 112 coupled to the lower end of the upper body cover 111 to form an inner space for the optical module ( 130) and the motor 140 is accommodated. At this time, a hole is formed in the center of the optical module 130 so that the space utilization inside the upper body 110 can be maximized, so that the motor 140 is disposed in the center hole of the optical module 130 . It is preferable to be

A fixing bracket 113 is provided at the lower end of the inner space formed by the upper body cover 111 and the upper and lower cover 112 , that is, at the upper side of the upper and lower cover 112 , The motor 140 is provided at the center of the upper surface. In addition, a rotation bracket 114 couples the rotation shaft of the optical module 130 and the motor 140 , and a slip ring 115 is provided on the rotation shaft of the motor 140 to supply power to the motor 140 . do. At this time, the upper and lower cover 112 and the fixing bracket 113 so that the optical module 130 does not block the progress of light in the process of performing light irradiation and detection for the sample accommodated in the tube 200 . At positions corresponding to the respective tubes 200, upper and lower cover through-holes 112h and fixing bracket through-holes 113h are formed.

In addition, the upper body 110 may further include a hot top heater substrate 116 provided between the fixing bracket 113 and the upper and lower cover 112 . The hot top heater substrate 116 is a device for uniformly heating the plurality of tubes 200 , and serves to assist the heating operation of the heater module 150 , which will be described later. At this time, the hot-top heater substrate 116 also has a hot-top heater substrate through-hole 116h formed at a position corresponding to the tube 200 so as not to block the propagation of light.

Meanwhile, as described above, the motor 130 and the optical module 140 are accommodated in the upper body 110. The motor 130 is fixedly provided while the optical module 140 is the motor ( 130) may be structurally unstable as it is only connected to the rotation shaft. In order to solve this problem, a spring 117 is provided between the lower end of the optical module 140 and the fixing bracket 113, so that the lower end of the optical module 140 can be flexibly supported by using a restoring force.

In addition, the upper body 110 and the lower body 120 are formed to be separated and coupled so that the tube 200 can be smoothly inserted or removed from the PCR apparatus 100, at this time the upper body ( 110) and the lower body 120 may be formed to be completely separated from each other, of course, but there may be a problem that the original positions of each part are separated when they are reconnected. Therefore, it is preferable that the upper body 110 and the lower body 120 are formed so as to be able to open and close by being hinged at one side as shown in FIG. 2 . In FIG. 4 , the shaft 118 is a component provided on this hinge.

As described above, the lower body 120 has the heater module 150 and the cooling fan 160 therein. 5 is an exploded perspective view of the lower body of the PCR apparatus of the present invention, and the detailed configuration of the lower body 120 will be described with reference to the cross-sectional view of FIG. 3 and the exploded perspective view of FIG.

The lower body 120 forms an inner space by combining the lower upper cover 121, the lower side cover 122, and the lower lower cover 123 to accommodate the heater module 150 and the cooling fan 160. do. In addition, a plurality of the tubes 200 are arranged in a circle at the upper end of the lower body 120 , so that light irradiation and detection from the upper body 110 can be performed smoothly. At this time, similarly to the upper body 110, a through hole is formed in the center of the heater module 150 to maximize the space utilization and air flow in the lower body 120, so that the cooling fan ( 160) is preferably formed so that the forced-blowing air is circulated through the central through-hole of the heater module 150.

A plurality of circularly arranged lower upper cover through holes 121h are formed in the lower upper cover 121 so that the tube 200 is inserted therein, and the lower upper cover 121 is located directly at the lower end of the upper body 120 . is placed on The lower upper cover 121 is coupled to the upper end of the lower side cover 122 , and the lower lower cover 123 is coupled to the lower end of the lower side cover 122 . The cooling fan 160 is fixedly provided at the center of the upper surface.

A heater fixing substrate 124 is provided at the upper end of the inner space formed by the lower upper cover 121, the lower side cover 122 and the lower lower cover 123, that is, below the lower upper cover 121, , the heater fixing substrate 124 is coupled to the upper surface of the heater module 140 to fix the heater module 140 . At this time, the heater fixed substrate 124 is formed with a heater fixed substrate through hole 124h at a position corresponding to the tube 200 so that the heater structure 155 can pass therethrough.

A plurality of lower surface vents 125 are formed in the center of the lower lower cover 123 , and a plurality of side vents 126 are formed at the upper end of the side of the lower side cover 122 . Accordingly, when the cooling fan 160 forcibly blows air, the air can be circulated between the inner space of the lower body 120 and the outside through the side vent 126 and the lower vent 125 . At this time, since it is preferable to draw air from a wider space, it is preferable that air is introduced into the side vent 126 and discharged through the lower vent 125 .

Additionally, the lower body 120 has a switch 127 for turning on/off the power and a switch cover 128 that covers and protects and fixes the switch, or a multi-color LED that displays the state according to the internal operation. An indicator 129 and the like may be further provided.

The optical module 130 includes a plurality of optical systems 132 that are radially arranged at equal intervals and individually perform light irradiation and detection, respectively, for light irradiation and detection of the sample accommodated in the tube 200 . carry out 6 is an exploded perspective view of the optical module of the PCR apparatus of the present invention. A detailed configuration of the optical module 130 will be described with reference to FIG. 6 as follows.

The optical module 130 includes an arrangement provided on the fixing plate 132, which is composed of a fixing plate 131 basically formed in a circular shape and a plurality of the optical systems 132 . At this time, as described above, the optical systems 132 are appropriately spaced apart from each other at equal radial intervals in a circle to prevent cross-talk between each other. The optical system 132 includes a light source for irradiating excitation light and a detection means for detecting fluorescence, respectively, so that light irradiation and detection can be performed individually.

The optical module 130 is also provided on the upper surface of the arrangement, and while supporting the upper surface of the arrangement, a first optical control board 133 for controlling light irradiation and detection of the optical system 132 and the first A second optical control board 134 is provided laminated on the upper surface of the optical control board 133 to control the operation in conjunction with the motor 140 in accordance with the light irradiation and detection operation of the optical system 132 . Accordingly, structurally, the optical module 130 has a lower surface supported by the fixing plate 131 and an upper surface of the array including the plurality of optical systems 132 arranged in a circle, and the upper surface is the first and second optical control substrates ( 133 and 134 to form a stable structure.

On the other hand, as described above, when the hot top heater substrate 116 for performing auxiliary heating is provided on the upper body 110 , there is a risk that unnecessary heat is applied to the optical module 130 . In order to prevent this problem, the fixing plate 131 is preferably made of an insulating material capable of blocking heat.

The motor 150 serves to rotate the optical module 130 so that light irradiation and detection are performed on the sample accommodated in each of the tubes 200 at a predetermined cycle. That is, while the motor 140 rotates the optical module 130 at a predetermined rotation speed, the optical system 132 performs light irradiation and detection on the sample accommodated in the tube 200 . The rotation speed of the motor 140 is adjusted according to a signal sampling rate.

As well shown in the drawings, the PCR apparatus 100 of the present invention performs sample detection for the samples accommodated in the plurality of tubes 200 . As described above, in the present invention, by providing a smaller number of the optical systems 132 than the tube 200, the distance between the optical systems 132 is sufficiently widened to fundamentally solve the signal interference problem. In addition, in the present invention, the optical module 130 is formed to be rotated by the motor 140, and as the optical system 132 rotates, light irradiation and detection are performed while passing the position of the tube 200. do. At this time, if the arrangement structure of the optical system 132 and the tube 200 and the rotation speed of the motor 140 are all known, at which point the optical system 132 is located in which tube 200, etc. Since it can be easily identified, it is possible to smoothly obtain a target material detection signal by only appropriately processing the signal. That is, in the present invention, by making the light irradiation and detection operation/advance operation to the next tube position be made continuously, the shock or dynamic instability of the optical system or the movement device actuator caused by the discontinuous operation in the conventional PCR apparatus as described above can be greatly reduced.

With reference to the configuration of the optical module 130 and the motor 140 described above, the sample detection method of the PCR device of the present invention will be described in stages as follows. The sample detection method of the PCR apparatus largely includes a module rotation step, a light irradiation step, and a photodetection step.

In the module rotation step, the motor 140 rotates the optical module 130 at a predetermined rotation speed. In this case, the rotation speed may be appropriately determined according to a desired signal sampling rate as described above. The time for the optical system 132 to pass the sample accommodated in each tube 200 varies according to the rotation speed of the motor 140 , but the sampling rate is the same. It is possible to obtain a signal with a high granularity. Therefore, it is possible to detect a fluorescence signal more accurately than when the rotation speed is high. However, if the rotation speed is too slow, the detection efficiency may be reduced. Therefore, it is sufficient to determine the appropriate optimal rotation speed while considering both the detection efficiency and the detection accuracy.

In the light irradiation step, the optical system 132 irradiates the excitation light to the sample accommodated in the tube 200 . When the sample is irradiated with excitation light, electrons are excited, the excited state is maintained for a certain period of time, and then a fluorescence photon is generated as it transitions to the ground state.

In the photodetection step, the optical system 132 detects a target material in the sample using fluorescence generated from the sample accommodated in the tube 200 .

At this time, the light irradiation step and the light detection step are performed in a state in which the optical system 132 is continuously rotating. Accordingly, the optical system 132 irradiating light to any one tube 200 may not necessarily detect fluorescence generated in the same tube 200 . However, as described above, if the arrangement structure of the optical system 132 and the tube 200 and the rotation speed of the motor 140 are all known, the optical system 132 is located in which tube 200 at a certain point in time. Since it is easy to understand whether the excitation light is irradiated to which tube 200 at any point in time through appropriate signal processing, and how much time elapses after that, the relationship such as whether the tube 200 generates fluorescence, etc. Since it can be easily grasped, the optical system 132 for irradiating light with respect to any one tube 200 and the optical system 132 for detecting light are not necessarily the same.

The heater module 150 includes a heater structure 155 provided to surround each of the tubes 200 and configured to individually heat the tubes 200 . 7 is an exploded perspective view of the heater structure of the PCR apparatus of the present invention. A detailed configuration of the heater structure 155 will be described with reference to FIG. 7 as follows.

The heater structure 155 is formed in the form of a metal pipe and is formed in the form of a tube accommodating part 155a into which the tube 200 is inserted and accommodated, and is provided in the form of a film to surround the side surface of the tube accommodating part 155a. and a film heater unit 155b for heating. Here, since the tube 200 for accommodating the sample is accommodated in the tube accommodating part 155a, the tube accommodating part 155a becomes a well. Also, although not shown in the drawing, the heater structure 155 further includes a temperature detecting unit attached to the tube receiving unit 155a to measure the temperature of the tube 200 in a contact manner.

As such, in the present invention, the heater structure 155 for directly heating the tube 200 has a structure such that a heater is provided in each of the plurality of tubes 200 . Accordingly, the heater module 155 is configured such that the heater module 150 uniformly heats the plurality of tubes 200 or individually drives the heater structure 155 to individually heat each tube 200 . All heating operations can be performed.

As described above, even in the conventional PCR apparatus, the operation of uniformly heating a plurality of tubes using a heating block or the like has been well implemented. However, since the same heat is transmitted to all of the plurality of tubes in this way, for example, when cooling efficiency is changed according to the air flow direction during cooling using air blowing, there is a risk that the temperature may be formed differently depending on the tube position. Specifically, for example, when cooling is performed by blowing air from one side of the tube array arranged in two dimensions to the other side, the tube disposed close to the air inlet side is well cooled by air at the same temperature as the outside temperature, but , the tube disposed close to the air discharge side is cooled by the air that has already absorbed heat and has a temperature higher than the outside temperature, so the cooling efficiency is lowered. As such, if the cooling is made non-uniformly and the heating is made uniformly, as a result, the temperature is formed differently for each tube position, and it is impossible to induce a correct PCR reaction.

However, in the present invention, the film heater unit 155b is provided for each tube 200 individually, and the heater module 150 can operate the plurality of film heater units 155b individually. . Therefore, even if the cooling efficiency is formed somewhat non-uniformly according to the air flow as described above, the temperature of all the tubes 200 is smoothly controlled by measuring the temperature of each tube 200 and individually adjusting the amount of heating heat accordingly. and can be easily controlled as desired.

As described above, the cooling fan 160 is provided at the center of a circle formed by the arrangement of the plurality of tubes 200 and serves to cool the tube by forcibly blowing air around the tube 200 . As previously described in the detailed configuration of the lower body 120, the cooling fan 160 forcibly blows air, so that air is introduced into the side vent 126 and discharged to the lower vent 125 is formed. At this time, since the plurality of tubes 200 are arranged in a circle, uniform cooling can be achieved for all the tubes 200 .

That is, in the PCR apparatus 100 of the present invention, since the air flow itself at the same temperature is formed in a form that absorbs heat while passing through all the tubes 200 once and is discharged at once, the conventional PCR described above As with the device, the air that has been warmed by absorbing heat while cooling one other tube does not cause another tube to cool again. That is, all the tubes 200 can be uniformly cooled regardless of their positions.

With reference to the configuration of the heater module 150 and the cooling fan 160 described above, the temperature control method of the PCR apparatus of the present invention will be described in stages as follows. The temperature control method of the PCR apparatus includes a temperature rising step, a temperature decreasing step, and a temperature control step.

In the temperature raising step, the heater module 150 uniformly heats the plurality of tubes 200 . As described above, since the PCR reaction occurs at a certain predetermined reference temperature depending on the sample, it is heated all at once to reach the reference temperature once.

In the temperature lowering step, the cooling fan 160 forcibly blows air to uniformly cool the plurality of tubes 200 . At this time, as described above, in the present invention, uniform cooling can be achieved in all the tubes 200 due to their structural characteristics. However, due to an unexpected cause, the cooling efficiency may vary slightly in each of the tubes 200, and in such a case, when heating is performed again at once, a problem occurs in that the temperature is formed differently for each tube. In the present invention, this problem is prevented by using the temperature control step as follows.

In the temperature control step, the heater module 150 individually drives the heater structure 155 provided in each tube 200 to maintain the temperature of each tube 200 at a predetermined reference temperature. individually heated to That is, even if the cooling efficiency is slightly different in each tube 200 due to an unexpected cause, individual temperature control independent of each other is possible because each tube 200 is individually heated. The temperature of the tube 200 can be adjusted as desired.

As such, by enabling individual temperature control of the tube 200 in the present invention, it is possible not only to cope with unexpected changes in cooling efficiency as described above, but also to further improve the utility of the PCR apparatus 100 . That is, in the prior art, since the entire tube 200 is heated and cooled at once, the overall reference temperature could be set to only one value. Therefore, only one type of sample can be reacted using one PCR device 100 couldn't However, in the present invention, since individual temperature control is possible for the plurality of tubes 200 , the reference temperature may be set differently according to the tubes 200 . Therefore, multiple types of samples can be reacted at once using one PCR device 100 , thereby maximizing the utility of the PCR device 100 .

The present invention is not limited to the above-described embodiments, and the scope of application is varied, and anyone with ordinary knowledge in the field to which the present invention pertains without departing from the gist of the present invention as claimed in the claims It goes without saying that various modifications are possible.

100: PCR device
110: upper body 111: upper body cover
112: upper and lower cover 112h: upper and lower cover through hole
113: fixing bracket 113h: fixing bracket through hole
114: rotation bracket 115: slip ring
116: hot top heater substrate 116h: hot top heater substrate through hole
117: spring 118: shaft
120: lower body
121: lower upper cover 121h: lower upper cover through hole
122: lower side cover 123: lower lower cover
124: heater fixed substrate 124h: heater fixed substrate through hole
125: bottom vent 126: side vent
127: switch 128: switch cover
129: indicator
130: optical module
131: fixed plate 132: optical system
133: first optical control board 134: second optical control board
140: motor
150: heater module 155: heater structure
155a: tube receiving part 155b: film heater part
160: cooling fan
200: tube

Claims (14)

A PCR device 100 that generates a polymerase chain reaction (PCR) reaction by heating and cooling the sample accommodated in the tube 200, and detects a target material in the sample using fluorescence generated by irradiating the sample with excitation light (100) In
upper body 110;
a lower body 120 in which a plurality of the tubes 200 are arranged in a circle at an upper end;
It is accommodated in the upper body 110, is radially arranged at equal intervals, and includes a plurality of optical systems 132 that individually perform light irradiation and detection, respectively, and light irradiation for the sample accommodated in the tube 200. and an optical module 130 for performing detection;
a motor 140 accommodated in the upper body 110 and configured to rotate the optical module 130 so that light irradiation and detection are performed with respect to the sample accommodated in each of the tubes 200 at a predetermined cycle;
a heater module 150 including a heater structure 155 accommodated in the lower body 120 and provided to surround each of the tubes 200 so as to be able to individually heat the tubes 200;
a cooling fan (160) accommodated in the lower body (120) and provided in the center of a circular arrangement of a plurality of the tubes (200) to forcibly blow air around the tube (200) to cool it;
A PCR device comprising a.
According to claim 1, wherein the heater structure (155),
A tube accommodating part (155a) formed in the form of a metal pipe and into which the tube 200 is inserted and accommodated;
A film heater unit (155b) formed in the form of a film and provided to surround the side surface of the tube receiving unit (155a) to perform heating,
A temperature detection unit attached to the tube receiving unit 155a to measure the temperature of the tube 200 in a contact manner.
A PCR device comprising a.
According to claim 1, wherein the heater module 150,
A through hole is formed in the center so that the air forcedly blown by the cooling fan (160) passes through the central through hole of the heater module (150) and circulates.
According to claim 1, wherein the PCR device 100,
When heating, the heater module 150 heats the plurality of tubes 200 uniformly or individually drives the heater structure 155 to individually heat each tube 200,
When cooling, the cooling fan 160 is a PCR apparatus, characterized in that formed to cool the plurality of tubes (200) uniformly.
In the temperature control method of the PCR apparatus 100 according to claim 1,
a temperature raising step in which the heater module 150 heats the plurality of tubes 200 uniformly;
a temperature lowering step in which the cooling fan 160 forcibly blows air to uniformly cool the plurality of tubes 200;
Temperature control in which the heater module 150 individually drives the heater structures 155 provided in each tube 200 to individually heat the temperature of each tube 200 to maintain a predetermined reference temperature step;
Temperature control method of a PCR device comprising a.
According to claim 1, wherein the optical module 130,
Fixed plate 131 formed in a circular shape,
an arrangement comprising a plurality of the optical systems 132 and provided on the fixing plate 132;
a first optical control board 133 provided on the upper surface of the arrangement to control the light irradiation and detection operation of the optical system 132 while supporting the upper surface of the arrangement;
A second optical control board 134 is provided stacked on the upper surface of the first optical control board 133 to control a link operation with the motor 140 in accordance with the light irradiation and detection operation of the optical system 132 .
A PCR device comprising a.
According to claim 1, wherein the optical module 130,
A through hole is formed in the center so that the motor 140 is disposed in the center through hole of the optical module 130. PCR device.
According to claim 1, wherein the PCR device 100,
The motor 140 rotates the optical module 130 at a predetermined rotation speed while the optical system 132 is formed to perform light irradiation and detection on the sample accommodated in the tube 200 PCR device.
The method of claim 8, wherein the PCR device 100,
PCR apparatus, characterized in that the rotation speed of the motor 140 is adjusted according to a desired signal segmentation rate (sampling rate).
In the sample detection method of the PCR apparatus 100 according to claim 1,
a module rotation step in which the motor 140 rotates the optical module 130 at a predetermined rotation speed;
a light irradiation step in which the optical system 132 irradiates excitation light to the sample accommodated in the tube 200;
a photodetection step in which the optical system 132 detects a target material in the sample using the fluorescence generated from the sample accommodated in the tube 200;
A sample detection method of a PCR device comprising a.
According to claim 1, wherein the PCR device 100,
PCR device, characterized in that the upper body 110 and the lower body 120 are formed so as to be able to open and close.
According to claim 1, wherein the upper body 110,
The upper body cover 111 forming a space inside,
The upper and lower cover 112 coupled to the lower end of the upper body cover 111 and having an upper and lower cover through hole 112h at a position corresponding to the tube 200;
A fixing bracket 113 that is provided on the upper side of the upper and lower cover 112, the motor 140 is fixed at the center of the upper surface, and a fixing bracket through hole 113h is formed at a position corresponding to the tube 200;
a rotation bracket 114 for coupling the optical module 130 and the rotation shaft of the motor 140;
Slip ring 115 for supplying power to the motor 140
A PCR device comprising a.
According to claim 12, The upper body 110,
It is provided between the fixing bracket 113 and the upper and lower cover 112 to uniformly heat a plurality of the tubes 200, and a hot top heater substrate through hole 116h is formed at a position corresponding to the tube 200. Hot Top Heater Substrate(116)
PCR device further comprising a.
According to claim 1, wherein the lower body 120,
A plurality of circularly arranged lower upper cover through-holes 121h are formed, the tube 200 is inserted, and the lower upper cover 121 is disposed at the lower end of the upper body 120,
The lower upper cover 121 is coupled to the upper end and the lower side cover 122 forming a space therein;
A lower lower cover 123 coupled to the lower end of the lower side cover 122 and having the cooling fan 160 fixed to the center of the upper surface;
It is provided on the lower side of the lower upper cover 121 and is coupled to the upper surface of the heater module 140 to fix the heater module 140, and a heater fixing substrate through hole 124h is formed at a position corresponding to the tube 200, A heater fixing substrate 124 formed so that the heater structure 155 is penetrated;
A plurality of lower surface vents 125 formed in the center of the lower lower cover 123 to distribute the air forcedly blown by the cooling fan 160 to the outside;
A plurality of side vents 126 formed on the upper side of the lower side cover 122 to distribute the air forcedly blown by the cooling fan 160 with the outside.
A PCR device comprising a.

Images