WO2022114814A1

WO2022114814A1 - Thermal module and method of operating same

Info

Publication number: WO2022114814A1
Application number: PCT/KR2021/017533
Authority: WO
Inventors: Jae Young Kim
Original assignee: Seegene, Inc.
Priority date: 2020-11-26
Filing date: 2021-11-25
Publication date: 2022-06-02
Also published as: KR20230088831A; WO2022114816A1; KR20230088830A

Abstract

Provided is a thermal module including a first thermal conductor to which a sample holder is to be thermally connected. The thermal module includes a first heating element thermally connected to the first thermal conductor and adapted to perform a heating operation to heating the sample holder. A themoelectric element is thermally connected to the first thermal conductor and adapted to perform a cooling operation to cool the sample holder.

Description

THERMAL MODULE AND METHOD OF OPERATING SAME

The present disclosure relates to a thermal module for a nucleic acid reaction and a method of operating the same.

A most commonly used nucleic acid amplification reaction that is well-known as a polynucleotide chain reaction (PCR) includes a repeated cycle process comprised of denaturation of a double-stranded DNA, annealing of an oligonucleotide primer to a DNA template, and primer extension by DNA polymerase (Mullis et al, U.S. Patent Nos. 4,683,195, 4,683,202, and 4,800,159; and Saiki et al., 1985, Science 230, PP. 1350-1354).

The denaturation of DNA proceeds at a temperature of about 95°C, and the annealing and the extension of the primer proceed at a temperature of 55°C to 75°C, which is lower than 95°C. Thus, the nucleic acid amplification reaction of samples is performed by repeating processes of raising and lowering temperatures of reaction containers or chambers in which the samples are contained. In this case, to heat or cool the samples, Peltier elements configured to be connected to the reaction containers or chambers are used. That is, in the denaturation operation of DNA, the samples are heated to about 95°C by the Peltier elements, and in the annealing and extension operations of the primer, the samples are cooled to about 55°C to 75°C by the Peltier elements.

Nucleic acid amplification devices for performing the nucleic acid amplification reaction often use the Peltier elements to heat or cool the samples. The Peltier elements have a limit in that cooling efficiency is lowered as a temperature difference between a high temperature section and a low temperature section becomes larger, and thus, have a problem in that efficiency thereof is low in spite of high power consumption.

For example, a Peltier element heating a sample in the denaturation operation has a great temperature difference between a heat radiating section and a heat absorbing section. Thus, the Peltier element heating the sample in the following operations of annealing and extending the primer has low cooling efficiency due to the temperature difference.

Therefore, there is a need to develop a heat exchange module having improved heat exchange efficiency, in particular, cooling efficiency, for performing a nucleic acid amplification reaction and a method of controlling the same.

To overcome the problems of the above related art, an objective of the present disclosure is to provide a thermal module including a heating element performing a heating operation to heat a sample holder and a themoelectric element performing a cooling operation to cool the sample holder, so that cooling efficiency of the themoelectric element can be secured in a cooling operation of the themoelectric element, and a method of operating the same.

Further, another objective of the present disclosure is to provide a thermal conductor to which the sample holder is to be thermally connected and to provide the heating element and the themoelectric element to be thermally connected to the thermal conductor, such that the heating operation of the heating element and the cooling operation of the themoelectric element are indirectly performed on the sample holder through the thermal conductor.

In addition, another objective of the present disclosure is to provide the themoelectric element to generate heat from a surface thereof thermally connected to a first thermal conductor during the heating operation of the heating element, thereby minimizing a loss of heat toward the themoelectric element during the heating operation of the heating element.

According to an embodiment of the present disclosure, a thermal module may include: a first thermal conductor to which a sample holder is to be thermally connected; a first heating element thermally connected to the first thermal conductor and adapted to perform a heating operation to heating the sample holder; and a themoelectric element thermally connected to the first thermal conductor and adapted to perform a cooling operation to cool the sample holder.

According to an embodiment of the present disclosure, provided is a method of operating a thermal module including a first thermal conductor to which a sample holder is to be thermally connected. The method may include the operations of: heating the sample holder by a first heating element thermally connected to the first thermal conductor; and cooling the sample holder by a themoelectric element thermally connected to the first thermal conductor.

The present disclosure provides the heating element performing a heating operation to heat the sample holder and a thermoelectric element performing a cooling operation to cool the sample holder, thereby having an effect capable of securing cooling efficiency of the thermoelectric element during the cooling operation of the thermoelectric element.

Further, the present disclosure is configured such that the heating operation of the heating element and the cooling operation of the thermoelectric element are indirectly performed on the sample holder through the thermal conductor, thereby having an effect capable of constructing a module in a compact way.

In addition, the present disclosure is configured such that, during the heating operation of the heating element, the thermoelectric element emits heat from a plane thermally connected to the first thermal conductor, thereby having an effect capable of minimizing a loss of heat toward the thermoelectric element during the heating operation of the heating element.

FIGS. 1 to 3 are configuration views illustrating a thermal module according to an embodiment of the present disclosure.

FIGS. 4 and 5 are perspective views illustrating the thermal module according to an embodiment of the present disclosure.

FIG. 6 is an exploded perspective view illustrating a part of the thermal module according to an embodiment of the present disclosure.

FIG. 7 is a cross-sectional view illustrating a part of the thermal module according to an embodiment of the present disclosure.

Hereinafter, the present disclosure will be described in more detail with reference to embodiments. It will be apparent to those having ordinary knowledge in the art that the following embodiments are for illustrative purposes only and the scope of the present disclosure is not limited by the embodiments.

In designating elements of the drawings by reference numerals, the same elements will be designated by the same reference numerals if possible although they are shown in different drawings. Further, in the following description of the present disclosure, a detailed description of known functions and configurations incorporated herein will be omitted in the situation in which the subject matter of the present disclosure may be rendered unclear thereby.

In addition, terms, such as "first", "second", "A", "B", "(A)", or "(B)" may be used herein to describe elements of the present invention. Each of these terms is not used to define essence, order, sequence, or number of elements, etc., but is used merely to distinguish the corresponding element from other elements. When it is mentioned that a first element is "connected", "coupled", or "linked" to a second element, it should be interpreted that, not only can the first element be directly connected, coupled, or linked to the second element, but a third element can also be "connected", "coupled", or "linked" between the first and second elements.

The term "sample" as used herein refers to a substance estimated including or supposed to include an analyte.

The "sample" may include biological samples (e.g., cells, tissue, and body fluid from biological supply sources) and non-biological samples (e.g., food, water, and soil). The biological samples may include, but are not limited to, virus, bacteria, tissue, cells, blood (e.g., whole blood, plasma, and serum), lymph, bone marrow fluid, saliva, sputum, swab, aspiration, milk, urine, feces, ocular fluid, semen, brain extract, spinal cord fluid (SCF), joint fluid, extracts from appendix, spleen and tonsil tissue, thymic fluid, bronchial lavage fluid, ascitic fluid, and amniotic fluid. Further, the samples may include natural nucleic acid molecules and synthetic nucleic acid molecules isolated from biological sources. According to an embodiment of the present disclosure, the "sample" may include substances used for preservation, processing, detection, etc. of the sample. The "sample" may include, but is not limited to, additional substances such as reagents for amplification, reagents for detection, preservatives, water, deionized water, saline water, pH buffer solutions, acid solutions, and basic solutions.

The term "sample processing" as used herein refers to a series of processes of primarily separating an analysis target substance from the sample and obtaining a substance from which a detection reaction is possible. The sample processing may be used as having a meaning that additionally includes a process of detecting a target analysis substance from the substance from which the detection reaction is possible. The analysis target substance may be, for example, nucleic acid.

The nucleic acid detection may be performed by a signal-generating reaction.

The term "signal-generating reaction" as used herein means any reaction capable of generating signals in a dependent manner on properties of a target analyte in a sample. The properties may be, for instance, activity, amount of presence (or absence) of the target analyte, in particular, the presence (or absence) of the target analyte in a sample. According to an embodiment of the present disclosure, the signal-generating reaction includes a biological reaction and a chemical reaction. The biological reaction includes a genetic analysis process such as PCR (Polymerase Chain Reaction), real-time PCR, microarray analysis and invader analysis, an immunological analysis process, and a bacterial growth analysis process. According to an embodiment of the present disclosure, the signal-generating reaction is a genetic analysis process. The chemical reaction includes the process of analyzing the creation, change or destruction of a chemical substance. According to an embodiment of the present disclosure, the signal-generating reaction is a signal amplification reaction.

The term "signal amplification reaction" as used herein means a reaction that increases or decreases a signal generated by the signal-generating means. According to an embodiment of the present disclosure, the amplification reaction means an increase (or amplification) reaction of a signal generated by the signal-generating means depending on the presence of a target analyte. The amplification reaction may or may not be accompanied by amplification of a target analyte (eg, a nucleic acid molecule). Particularly, in the present disclosure, the amplification reaction means a signal amplification reaction accompanied by amplification of a target analyte.

There have been known various methods of generating an optical signal indicating presence of a target nucleic acid using a signal-generating reaction. Representative examples of the various methods are as follows: a PTO cleavage and extension (PTOCE) method (WO 2012/096523), a TaqMan^TM probe method (U.S. Pat. No. 5,210,015), a molecule beacon method (Tyagi et al., Nature Biotechnology v. 14 MARCH 1996), a scorpion method (Whitcombe et al., Nature Biotechnology 17: 804-807 1999)), a sunrise or amplifluor method (Nazarenko et al., Nucleic Acids Research, 2512: 2516-2521 1997) and U.S. Pat. No. 6,117,635), a lux method (U.S. Pat. No. 7,537,886), a CPT (Duck P, et al., Biotechniques, 9: 142-148 1990)), an LNA method (U.S. Pat. No. 6,977,295), a Plexor method (Sherrill C B, et al., Journal of the American Chemical Society, 126: 4550-4556 (2004)), Hybeacons^TM (D. J. French, et al., Molecular and Cellular Probes (2001) 13, 363-374 and U.S. Pat. No. 7,348,141), a dual-labeled self-quenched probe (U.S. Pat. No. 5,876,930), a hybridization probe (Bernard P.S., et al., Clin Chem 2000; 46, 147-148), Detection of target nucleic acid sequences by PTO cleavage and extension (PTOCE) assay (WO2012/096523), Detection of target nucleic acid sequence by PTO cleavage and extension-dependent signaling oligonucleotide hybridization (PCE-SH) assay (WO2013/115442), Detection of target nucleic acid sequence by PTO cleavage and extension-dependent non-hybridization (PCE-NH) assay (PCT/KR2013/012312), Detection of target nucleic acid sequences by cyclic exonucleolytic reactions (CER) (WO2011/037306), and Assimilating probe method (PCT/US2011/041540).

Various types of nucleic acid amplification reactions may be performed using the thermal module of the present procedure and the method of operating the same. For example, the nucleic acid amplification reactions include polymerase chain reaction (PCR), ligase chain reaction (LCR) (U.S. Patent Nos. 4,683,195 and 4,683,202; PCR Protocols: A Guide to Methods and Applications (Innis et al., eds, 1990)), Strand displacement amplification (SDA) (Walker, et al. Nucleic Acids Res. 20(7): 1691-6, 1992); Walker PCR Methods Appl. 31): 1-6, 1993)), transcription-mediated amplification (Phyffer, et al., J. Clin. Microbiol. 34: 834-841, 1996); and Vuorinen, et al., J. Clin. Microbiol. 33: 1856-1859, 1995)), Nucleic acid sequence-based amplification (NASBA) (Compton, Nature 350 (6313): 91-2 ,1991)), Rolling circle amplification (RCA) (Lisby, Mol. Biotechnol. 121): 75-99, 1999), Hatch et al., Genet. Anal. 15(2): 35-40, 1999)), Q-beta Replicase (Lizardi et al., BiolTechnology 6: 1197, 1988)), loop-mediated isothermal amplication (LAMP, Y. Mori, H. Kanda and T. Notomi, J. Infect. Chemother., 2013, 19, 404-411), and recombinase polymerase amplication(RPA, J. Li, J. Macdonald and F. von Stetten, Analyst, 2018, 144, 31-67), etc.

Especially, a thermal module of the present disclosure and a method of operating the same are put to good use in a nucleic acid amplification reaction based on a polymerase chain reaction. Various nucleic acid amplifying methods using a polymerase chain reaction have been known. For example, the various nucleic acid amplifying methods include quantitative PCR, digital PCR, asymmetric PCR, reverse transcription polymerase chain reaction (RT-PCR), differential display PCR (DD-PCR), nested PCR, arbitrarily primed polymerase chain reaction (AP-PCR), multiplex PCR, SNP genome typing PCR, etc.

The term "cycle" as used herein means a unit of changes of conditions in a plurality of measurements accompanied with changes of conditions. The change of the conditions means, for example, an increase or decrease in temperature, reaction time, reaction number, concentration, pH, and replication number of a measurement target (e.g., nucleic acid). Thus, a cycle may be a time or process cycle, a unit operation cycle or a reproductive cycle.

As one example, when analyzing a substrate decomposition capacity of an enzyme according to a substrate concentration, the substrate decomposition capacity of the enzyme is analyzed therefrom after measuring the substrate decomposition degree of the enzyme several times by varying the substrate concentration. At this time, the change of the condition is an increase in the substrate concentration, and the used substrate concentration increase unit is set as one cycle.

As another example, when performeing isothermal amplification of nucleic acid, several measurement may be performed with different reaction times for a sample. In this case, the reaction time is the change of the condition, and the reaction time unit is set as one cycle. For example, the measurement may be perforemed several times with changing the reaction time, such as 1 minute, 2 minutes, 3 minutes, etc. In this case, a cycle has a unit of time, and one cycle is set as a reaction time unit of 1 minute.

More specifically, the term "cycle" may refer to one unit of repetition when a reaction of a constant process is repeated or when a reaction is repeated on the basis of a fixed time interval.

For example, in the case of the polymerase chain reaction (PCR), one cycle refers to a reaction including an operation of denaturing nucleic acid, an operation of binding (i.e., hybridizing or annealing) the nucleic acid and a primer, and an operation of extending the primer. In this case, a constant change in conditions includes an increase in the number of repetitions of reactions, and a unit of repetitions of reactions including the series of operations is set as one cycle.

FIGS. 1 to 3 are configuration views illustrating a thermal module according to an embodiment of the present disclosure. A configuration and arrangement of the thermal module according to an embodiment of the present disclosure are illustrated in FIGS. 1 to 3 relative to a sample holder 10. FIGS. 4 to 7 are perspective views and a cross-sectional view illustrating the thermal module according to an embodiment of the present disclosure.

A sample is held in a sample holder 10, and the sample holder 10 may be, for example, a tube, a vessel, or a cuvette. A target nucleic acid is included in the sample held in the sample holder 10 in a state in which the target nucleic acid may be subjected to a detection reaction by a sample processing process. The sample holder 10 includes a container portion 11 in which the sample is held, and may further include a heat conduction portion 12. Although an embodiment in which the sample holder 10 includes the container portion 11 and the heat conduction portion 12 thermally connected to a first thermal conductor 110 is illustrated in the figures, the embodiment is not necessarily limited thereto. For example, the sample holder 10 may include only the container portion or include the container portion, the heat conduction portion thermally connected to the first thermal conductor 110, and the heat conduction portion thermally connected to a second thermal conductor 150. In addition, a sample space in which the sample is held is provided in one surface of the container portion 11 by carving, and the heat conduction portion 12 may be coupled to cover the sample space on one surface of the container portion 11. For example, the container portion 11 may be provided in a hexahedral form, the sample space may be provided in one among six surfaces by carving, and the heat conduction portion 12 may be coupled to cover the sample space in the one surface. According to this structure, the sample held in the sample space can be directly thermally connected to the heat conduction portion 12, so that the sample can be heated more rapidly. The heat conduction portion of the sample holder 10 may be made of a metal layer, for example, a metal layer made of aluminum.

First, a description will be made with reference to FIG. 1.

A thermal module 100 of the present disclosure heats or cools the sample holder 10, so that an amplification reaction of the target nucleic acid included in the sample is performed. The thermal module 100 includes a first thermal conductor 110 to which the sample holder 10 is to be thermally connected, a first heating element (or a heating element) 120 thermally connected to the first thermal conductor 110 to perform a heating operation to heat the sample holder 10, and a themoelectric element 130 thermally connected to the first thermal conductor 110 to perform a cooling operation to cool the sample holder 10.

The first heating element 120 is a component that performs the heating operation to heat the sample holder 10. The first heating element 120 may be a resistance heating module, for example, a resistance heating module of 12 V and 30 W. The first heating element 120 may be appropriately selected from 12V/24V and 10W/20W/30W/40W/50W/60W. The themoelectric element 130 is a component that performs the cooling operation to cool the sample holder 10, and the themoelectric element 130 may be a Peltier element or a thermoelectric cooler (TEC). The themoelectric element 130 is thermally connected to the first thermal conductor 110 and a heat sink 142, and may be thermally connected to the heat sink 142 via a metal block 141. In addition, the thermal module 100 of the present disclosure has a temperature sensor for measuring temperatures of the sample holder 10 and/or the first thermal conductor 110, and thus, can detect whether the sample holder 10 is heated or cooled at an appropriate temperature. The temperature sensor may be provided for a thermal pusher 400 (to be described below).

The sample holder 10, the first heating element 120, and the themoelectric element 130 are thermally connected to the first thermal conductor 110. The expression "thermally connected" means that heat may be mutually exchanged. A thermal conductor for mediating heat exchange may be additionally included. For example, the sample holder 10, the first heating element 120, or the themoelectric element 130 may be thermally contacted to the first thermal conductor 110.

Further, heating of the sample holder 10 based on the first heating element 120 and cooling of the sample holder 10 based on the themoelectric element 130 are indirectly performed via the first thermal conductor 110. That is, the first heating element 120 and the themoelectric element 130 are indirectly thermally connected to the sample holder 10 via the first thermal conductor 110 without being directly thermally connected to the sample holder 10.

The first thermal conductor 110 includes two planes, and the sample holder 10, the first heating element 120, and the themoelectric element 130 may be connected to the two planes. To compactly dispose the sample holder 10, the first heating element 120, and the themoelectric element 130 with respect to the first thermal conductor 110, some of the sample holder 10, the first heating element 120, and the themoelectric element 130 may be thermally connected to one of the two planes, and the other thereof may be thermally connected to the other of the two planes. The first heating element 120 and the themoelectric element 130 may be thermally connected to any one of the different planes of the first thermal conductor 110. According to an embodiment illustrated in the figure, the themoelectric element 130 may be thermally connected to any one of the two planes of the first thermal conductor 110, while the sample holder 10 and the first heating element 120 may be thermally connected to the other of the two planes of the first thermal conductor 110. According to another embodiment, the sample holder 10 may be thermally connected to any one of the two planes of the first thermal conductor 110, while the first heating element 120 and the themoelectric element 130 may be thermally connected to the other of the two planes of the first thermal conductor 110. According to another embodiment, the first heating element 120 may be thermally connected to any one of the two planes of the first thermal conductor 110, while the sample holder 10 and the themoelectric element 130 may be thermally connected to the other of the two planes of the first thermal conductor 110. According to another embodiment, the sample holder 10, the first heating element 120, and the themoelectric element 130 may be thermally connected to any one of the two planes of the first thermal conductor 110, and heat exchange of the other of the two planes of the first thermal conductor 110 may be additionally interrupted by an insulator. In the embodiments, to prevent heat loss of the first thermal conductor 110, each of the two planes is preferably covered by the sample holder 10, the first heating element 120, the themoelectric element 130, or the insulator. Further, in any of the embodiments in which any two of the sample holder 10, the first heating element 120, and the themoelectric element 130 are thermally connected to the same plane of the first thermal conductor 110, any two thereof may be connected to the first thermal conductor 110 in the same or different region. Further, in any of the embodiments in which the sample holder 10, the first heating element 120, and the themoelectric element 130 are thermally connected to the same plane of the first thermal conductor 110, all thereof may be connected to the first thermal conductor 110 in the same region, whereas any two thereof may be connected in the same region, and the other one may be connected in different regions, or all thereof may be connected in different regions.

The two planes of the first thermal conductor 110 may be a top plane and a bottom plane. On the basis of the embodiment illustrated in the figure, the themoelectric element 130, the first thermal conductor 110, the sample holder 10, and the first heating element 120 are layered up and down in this order. For example, the first heating element 120 may be thermally connected to the top plane of the first thermal conductor 110, and the sample holder 10 and the themoelectric element 130 may be thermally connected to the bottom plane of the first thermal conductor 110. Alternatively, the two planes of the first thermal conductor 110 may be a front plane and a back plane. The front and back sides correspond to one side and the other side in a horizontal direction (or a lateral direction), and are not particularly limited. For example, as will be described below, the front and back sides may be a direction in which the pusher 400 slides toward the sample holder 10 and the opposite direction. On the basis of the embodiment illustrated in the figure, the themoelectric element 130, the first thermal conductor 110, the sample holder 10, and the first heating element 120 may be disposed in this order in a horizontal (or lateral) direction. For example, the sample holder 10 and the first heating element 120 may be thermally connected to the front plane of the first thermal conductor 110, and the themoelectric element 130 may be thermally connected to the back plane of the first thermal conductor 110.

The first thermal conductor 110 may be implemented as a metal layer, so that heat exchange between the sample holder 10 and the first heating element 120 and between the sample holder 10 and the themoelectric element 130 can be rapidly performed. The first thermal conductor 110 is implemented as a metal layer having low heat capacity and a wide surface area, so that the heating or cooling of the sample holder 10 can be rapidly performed during a heating operation caused by the first heating element 120 and a cooling operation caused by the themoelectric element 130. This metal layer is made of a metal having high thermal conductivity, for example, aluminum (Al), copper (Cu), silver (Ag), or combinations thereof, an Al-Mg alloy, an Al-Si alloy, gold (Au), or tungsten (W). In addition, a plane of the themoelectric element 130 which is thermally connected to the first thermal conductor 110 and/or a plane thermally connected to the metal block 141 may be covered with a layer 170 including nano silver particles (or a nano silver layer) (see FIG. 3). In this case, the first thermal conductor 110 may be a metal layer made of silver.

During the heating operation of the first heating element 120 for heating the sample holder 10, the themoelectric element 130 may emit heat from the plane thermally connected to the first thermal conductor 110. Thus, it is possible to prevent or reduce heat loss caused when the heat generated by the first heating element 120 and transferred to the first thermal conductor 110 is conducted to the themoelectric element 130 rather than the sample holder 10. That is, the denaturation operation during the amplification reaction of the target nucleic acid included in the sample holder 10 may be performed due to the heating operation of the first heating element 120. In this case, the themoelectric element 130 can generate heat from the plane thermally connected to the first thermal conductor 110, thereby preventing or reducing heat loss caused when the heat generated by the first heating element 120 is conducted to the themoelectric element 130 rather than the sample holder 10. Preferably, the heating of the sample holder 10 for performing the denaturation operation is performed by the heating operation of the first heating element 120 rather than generating heat from the themoelectric element 130, and the generation of heat from the themoelectric element 130 is for preventing or reducing the generation of heat loss from the themoelectric element 130.

A thermal module according to the related art has performed both heating and cooling the sample using one Peltier element. However, after the Peltier element performs the denaturation operation at a temperature of about 95°C, a temperature difference between high and low temperature sections of the Peltier element becomes greater. Thus, in the case in which the Peltier element cools the sample to perform the annealing and extension operations of the primer between about 55°C and 75°C after the denaturation operation, there is a problem in which cooling efficiency of the Peltier element is very low due to the temperature difference.

However, the thermal module 100 of the present disclosure includes the first heating element 120 and the themoelectric element 130, such that the denaturation operation is performed by the heating operation of the first heating element 120, and the themoelectric element 130 is operated to generate heat from the plane thermally connected to the first thermal conductor 110 in order to prevent or reduce generation of heat loss during the heating operation of the first heating element 120. Consequently, even after the denaturation operation is performed, the temperature difference between the high and low temperature sections of the themoelectric element 130 is smaller compared to the related art, and thus, the themoelectric element 130 can perform the cooling operation to cool the sample holder 10 with relatively high cooling efficiency.

According to another embodiment of the present disclosure, during the heating operation of the first heating element 120, the thermoelectric element 130 may heat a plane thereof thermally connected to the first thermal conductor 110 to the extent that the cooling operation after the heating operation is enhanced.

According to an embodiment of the present disclosure, during the heating operation, the thermoelectric element 130 may heat the first thermal conductor 110 to a temperature equal to or lower than the temperature of the first heating element 120.

According to an embodiment of the present disclosure, during the heating operation, the thermoelectric element 130 may heat the first thermal conductor 110 to 95° C. or less.

According to another embodiment of the present disclosure, during the heating operation, the thermoelectric element 130 may heat the first thermal conductor 110 to 80° C. or less.

According to another embodiment of the present disclosure, during the heating operation, the thermoelectric element 130 may heat the first thermal conductor 110 to 60° C. or less.

According to an embodiment of the present disclosure, the temperature of the first heat conductor 110 heated by the thermoelectric element 130 may be changed during the heating operation.

Further, during the cooling operation of the themoelectric element 130, the first heating element 120 stops the heating operation. The themoelectric element 130 absorbs heat from the plane connected to the first thermal conductor 110 and cools the sample holder 10. As the heating operation of the first heating element 120 is stopped, the themoelectric element 130 can rapidly perform the cooling of the sample holder 10. That is, due to the cooling operation of the themoelectric element 130, the annealing and extension operations of the primer may be performed during the amplification reaction of the target nucleic acid included in the sample of the sample holder 10. The cooling operation of the themoelectric element 130 may be performed after the denaturation operation of the target nucleic acid is performed by the heating operation of the first heating element 120. As described above, the temperature difference between the high and low temperature sections of the themoelectric element 130 after the denaturation operation is performed is smaller than that between the high and low temperature sections of the Peltier element after the denaturation operation of the thermal module is performed. Thus, the themoelectric element 130 can perform the cooling operation to cool the sample holder 10 with relatively high cooling efficiency.

Further, the thermal module 100 of the present disclosure may further include a heat radiator 140 to emit the heat absorbed from the first thermal conductor 110 of the themoelectric element 130. In an embodiment, the heat radiator 140 may include a heat sink 142 to absorb and emit the heat generated by the cooling operation of the themoelectric element 130 and a ventilating fan 143 to cool the heat sink 142. The heat sink 142 is provided to be thermally connected to the themoelectric element 130 to absorb the heat.

In another embodiment, the heat radiator 140 may further include a metal block 141 that thermally connects the themoelectric element 130 and the heat sink 142. An embodiment in which the metal block 141 is thermally connected to the themoelectric element 130 on the opposite side of the first thermal conductor 110 is illustrated in the figure.

Next, a description will be made with reference to FIG. 2A.

The thermal module 100 of the present disclosure may further include a second thermal conductor 150 that is thermally connected to the sample holder 10, and a second heating element 160 that is thermally connected to the second thermal conductor 150 and performs a heating operation to heat the sample holder 10.

Like the first heating element 120, the second heating element 160 is a component for performing the heating operation of heating the sample holder 10. The second heating element 160 may be a resistance heating module, for example, a resistance heating module of 12V and 10W. The second heating element 160 may be appropriately selected among from 12V/24V and 10W/20W/30W/40W/50W/60W. Each of the sample holder 10 and the second heating element 160 is thermally connected to the second thermal conductor 150. For example, the second heating element 160 may be thermally contacted to the second thermal conductor 150. The heating of the sample holder 10 caused by the second heating element 160 is indirectly performed via the second thermal conductor 150. That is, the second heating element 160 is not directly thermally connected to the sample holder 10, but is indirectly thermally connected via the second thermal conductor 150. The second thermal conductor 150 may be implemented as a metal layer, so that heat exchange between the sample holder 10 and the second heating element 160 may be rapidly performed. The second thermal conductor 150 is implemented as a metal layer having low heat capacity and a wide surface area. The sample holder 10 may be rapidly heated during the heating operation caused by the second heating element 160. This metal layer is made of a metal having high thermal conductivity, for example, aluminum (Al), copper (Cu), silver (Ag), or combinations thereof, or an Al-Mg alloy, an Al-Si alloy, gold (Au), or tungsten (W).

The first thermal conductor 110 and the second thermal conductor 150 may be thermally connected to different surfaces of the sample holder 10. As illustrated in the figure, the first thermal conductor 110 and the second thermal conductor 150 may be thermally connected to the mutually opposite surfaces of the sample holder 10.

The second heating element 160 may heat the sample holder 10 along with the first heating element 120. That is, the heating operation of the first heating element 120 and the heating operation of the second heating element 160 may be performed at the same time. For example, to perform the denaturation operation, the heating operation of the first heating element 120 and the heating operation of the second heating element 160 may be performed at the same time. The heating of the sample holder 10 caused by the heating operation of the second heating element 160 may be additionally performed with respect to the heating of the sample holder 10 caused by the heating operation of the first heating element 120. That is, the first heating element 120 may be operated as a main heater of the sample holder 10, while the second heating element 160 may be operated as an auxiliary heater of the sample holder 10.

Further, during the cooling operation of the themoelectric element 130, the second heating element 160 may heat the sample holder 10 at a temperature lower than that at which the sample holder 10 is heated during the heating operation. That is, the second heating element 160 may perform a heating operation to heat the sample holder 10 along with the heating operation of the first heating element 120 and heat the sample holder 10 during the cooling operation of the themoelectric element 130. For example, in the case in which the themoelectric element 130 performs the cooling operation to cool the sample holder 10 in order to perform the annealing and extension operations of the primer after the denaturation operation is performed, the second heating element 160 may heat the sample holder 10 at a temperature lower than a temperature at which the sample holder 10 is heated during a heating operation performed at the same time as the heating operation of the first heating element 120 in the denaturation operation. Because the denaturation operation is performed at a temperature of about 95°C, the annealing and extension operations are performed at a temperature from about 55°C to 75°C. Thus, when the sample holder 10 is cooled by the themoelectric element 130, the second heating element 160 may heat the sample holder 10 so as to maintain the temperature for the sample holder 10.

According to an embodiment of the present disclosure, as illustrated in FIGS. 1, 2A, and 3, the thermal module 100 may be provided in such a form that the sample holder 10 may be inserted in a vertical direction.

Further, according to an embodiment of the present disclosure, as illustrated in FIG. 2B, the thermal module 100 may be provided in such a form that the sample holder 10 may be inserted in a horizontal direction.

FIG. 2B illustrates a different embodiment of the thermal module 100 including the same components as in FIG. 2A. However, respective components are configured in a layered form such that the sample holder 10 having a plane form may be located in the upper portion of the first thermal conductor 110. Thus, the sample holder 10 has a flat form, such that the second heating element 160 and the second thermal conductor 150 may be located at the upper portion of the sample holder 10.

Further, the thermal module 100 illustrated in FIGS. 1 and 3 may also be configured in a layered form so as to be able to use the sample holder 10 having a plane form as illustrated in FIG. 2B.

Next, a description will be made with reference to FIGS. 4 and 5.

The thermal module 100 of the present invention may further include the thermal pusher 400. The sample holder 10 is brought into close contact with the first thermal conductor 110 and the second thermal conductor 150 by the thermal pusher 400. The first thermal conductor 110 includes a surface facing the second thermal conductor 150 and the opposite surface thereof. The sample holder 10, the first heating element 120, and the themoelectric element 130 are thermally connected to the two surfaces.

In a state in which the sample holder 10 is in close contact with the first thermal conductor 110 and the second thermal conductor 150, the first heating element 120, the second heating element 160 and/or the themoelectric element 130 are operated and perform heating or cooling of the sample holder 10.

An embodiment in which the sample holder 10 is provided between the first thermal conductor 110 and the second thermal conductor 150 to face the first thermal conductor 110 and the second thermal conductor 15 in a horizontal direction is illustrated in the figures, but is not limited thereto. Rather, the sample holder 10 may be provided to face the first thermal conductor 110 and the second thermal conductor 15 in a vertical direction. The thermal pusher 400 includes a support 430 provided to be movable in a direction toward the first thermal conductor 110 and in the opposite direction thereof by a motor, a gear, a linear guide, etc. The second thermal conductor 150 is provided on a surface of the support 430 which faces the first thermal conductor 110. Thus, the sample holder 10 located between the first thermal conductor 110 and the second thermal conductor 150 may be brought into close contact with the first thermal conductor 110 and the second thermal conductor 150 by movement of the support 430. A movement structure based on the motor, the gear, the linear guide, etc. is the same as known in general, and thus, detailed description thereof will be omitted. A drive unit for moving the support 430 may be provided in a variety of configurations other than the structure using the motor, the gear, the linear guide, etc. That is, the drive unit may be configured by proper selection among, for example, a hydraulic/pneumatic piston, a rail, a belt, a chain, a pulley, a linear actuator, a rack and pinion, etc., to which the present disclosure is not limited.

The thermal module 100 may be provided with a control module for controlling the first heating element 120, the second heating element 160, the themoelectric element 130, and the drive unit. According to an embodiment, the control module may be implemented as a printed circuit board (PCB) (see reference numerals 500 and 510).

The sample holder 10 may be inserted from up to down and be located between the first thermal conductor 110 and the second thermal conductor 150, or inserted in a lateral direction and be located between the first thermal conductor 110 and the second thermal conductor 150. The sample holder 10 may perform sample processing after being located between the first thermal conductor 110 and the second thermal conductor 150, and then, a nucleic acid amplification reaction may be performed.

The thermal module 100 may include a guide for supporting the sample holder 10 located between the first thermal conductor 110 and the second thermal conductor 150. According to an embodiment, the guide may include a first guide 410 coupled to the metal block 141 and a second guide 420 coupled to the support 430. An embodiment in which the first guide 410 is supported on the lower surface and one lateral surface of the sample holder 10 and the second guide 420 is supported on the other lateral surface of the sample holder 10 is illustrated in the figures. A space in which the sample holder 10 may be located is defined by the first guide 410 and the second guide 420 so as to be open upward between the first thermal conductor 110 and the second thermal conductor 150. Unlike as illustrated in the figure, the first guide 410 may be supported on the lower surface (or the top surface) and one side (or the other side) of the sample holder 10, and the second guide 420 may be supported on the upper surface (the bottom surface) of the sample holder 10, and a space in which the sample holder 10 may be located between the first thermal conductor 110 and the second thermal conductor 150 may be provided to be open to the other side (or one side) by the first guide 410 and the second guide 420.

Next, description will be made with reference to FIG. 6.

The themoelectric element 130, the first thermal conductor 110, the first heating element 120, and the first guide 410 may be coupled to the metal block 141 in this order on the basis of the metal block 141. The sample holder 10 and the first heating element 120 may be located on the front surface facing the second thermal conductor 150 of the first thermal conductor 110의 second thermal conductor 150, and the themoelectric element 130 may be located on the back surface that is on the opposite side thereof. A portion of the front surface of the first thermal conductor 110 is thermally connected to the first heating element 120, and the other of the first thermal conductor 110 is exposed toward the second thermal conductor 150 and is thermally connected to the sample holder 10. The first guide 410 may cover the first heating element 120 and be coupled to the metal block 141.

The second heating element 160 and the second thermal conductor 150 may be coupled to the support 430 in this order. The sample holder 10 may be located on the front surface facing the first thermal conductor 110 of the second thermal conductor 150, and the second heating element 160 may be located on the rear face that is on the opposite side thereof. The front surface of the second thermal conductor 150 is exposed toward the first thermal conductor 110, and is thermally connected to the sample holder 10. The second guide 420 may be coupled to a lateral surface of the support 430.

Next, a description will be made with reference to FIG. 7.

A hole 440 passing through the support 430, the second heating element 160 and the second thermal conductor 150 may be provided in the thermal pusher 400. The hole 440 is open toward the first thermal conductor 110, particularly at a position corresponding to a position in which the sample of the sample holder 10 is held. An optical fiber (not illustrated) is inserted into the hole 440 of the thermal pusher 400 toward the first thermal conductor 110, excitation light generated by a light source by the optical fiber is provided to the sample, and emission light generated by the sample may be guided to a photodetector. Thus, when one cycle of performing operations of the nucleic acid amplification process, i.e., a denaturation operation, a primer annealing operation, and a primer extension operation, is completed, amplified nucleic acid can be detected in real time.

Next, a method of operating the thermal module of the present disclosure will be described. The method of operating the thermal module of the present disclosure is an operating method for heating the sample holder 10, or a method of operating the thermal module 100 which includes the first thermal conductor 110 to be in contact with the sample holder 10.

The method of operating the thermal module includes a heating operation in which the first heating element 120 thermally connected to the first thermal conductor 110 heats the sample holder 10, and a cooling operation in which the themoelectric element 130 thermally connected to the first thermal conductor 110 cools the sample holder 10. An amplification reaction of the target nucleic acid included in the sample of the sample holder 10 may be performed by the method of operating the thermal module. The sample holder 10 may be heated in the heating operation, and the denaturation operation during the amplification reaction may be performed. The sample holder 10 may be cooled, and the annealing operation and the extension operation of the primer during the amplification reaction may be performed.

In the heating operation, the themoelectric element 130 thermally connected to the first thermal conductor 110 may generate heat from the themoelectric element 130 thermally connected. There, when the first heating element 120 heats the sample holder 10, generated heat is conducted to the themoelectric element 130, so that generation of heat loss can be prevented or reduced.

In the cooling operation, the first heating element 120 may stop heating the sample holder 10. Thus, in the cooling operation, the cooling of the sample holder 10 can be more rapidly cooled by the cooling operation of the themoelectric element 130.

According to an embodiment of the present disclosure, the operation of the thermal module may be repeatedly performed a plurality of times by the number of cycles of the heating operation and the cooling operation. That is, one cycle of performing each step of the nucleic acid amplification process, i.e., the denaturation operation, the primer binding operation, and the extension operation, includes the heating operation and the cooling operation. As a result, since the nucleic acid amplification is performed by performing a plurality of cycles, the thermal module may repeat the heating operation and the cooling operation a plurality of times.

The first thermal conductor 110 includes two planes, and the sample holder 10, the first heating element 120, and the themoelectric element 130 may be thermally connected to the two plans. The two planes may be a top plane and a bottom plane. The two planes may be a front plane and a back plane. The first heating element 120 and the themoelectric element 130 may be thermally connected to different planes of the first thermal conductor 110.

Further, the thermal pusher 400 further includes the second thermal conductor 150 that is thermally connected to the sample holder 10. In the heating operation, the second heating element 160 thermally connected to the second thermal conductor 150 may heat the sample holder 10.

The first heating element 120 and the second heating element 160 may heat the sample holder 10 at the same time. In this case, the first heating element 120 may operate as a main heater, and the second heating element 160 may operate as an auxiliary heater.

In the cooling operation, the second heating element 160 can heat the sample holder 10 at a temperature lower than a temperature at which the sample holder 10 is heated in the heating operation.

[CROSS-REFERENCE TO RELATED APPLICATIONS]

This application claims priority from Korean Patent Application No. 10-2020-0161328, filed on November 26, 2020, and No. 10-2020-0187060, filed on December 30, 2020, in the Korean Intellectual Property Office, the disclosure of which is incorporated herein by reference in its entirety.

Claims

A thermal module comprising:

a first thermal conductor to which a sample holder is to be thermally connected;

a first heating element thermally connected to the first thermal conductor and adapted to perform a heating operation to heat the sample holder; and

a thermoelectric element thermally connected to the first thermal conductor and adapted to perform a cooling operation to cool the sample holder.
The thermal module according to claim 1, wherein, during the heating operation of the first heating element, the thermoelectric element heats a plane thereof thermally connected to the first thermal conductor to the extent that the cooling operation after the heating operation is enhanced.
The thermal module according to claim 1, wherein, during the cooling operation of the thermoelectric element, the first heating element stops the heating operation.
The thermal module according to claim 1, wherein the first thermal conductor comprises two planes, and

the sample holder, the first heating element, and the thermoelectric element are thermally connected to the two planes.
The thermal module according to claim 4, wherein the two planes include a top plane and a bottom plane.
The thermal module according to claim 4, wherein the two planes include a front plane and a back plane.
The thermal module according to claim 4, wherein the first heating element and the thermoelectric element are thermally connected to different planes of the first thermal conductor.
The thermal module according to claim 1, wherein the first thermal conductor comprises a metal layer.
The thermal module according to claim 8, wherein the metal layer comprises aluminum, copper, silver or combinations thereof.
The thermal module according to claim 1, wherein the first heating element is a resistance heating module.
The thermal module according to claim 1, further comprising:

a second thermal conductor thermally connected to the sample holder; and

a second heating element thermally connected to the second thermal conductor and adapted to perform the heating operation to heating the sample holder.
The thermal module according to claim 1, wherein the heating operation of the first heating element and the heating operation of the second heating element are performed at the same time.
The thermal module according to claim 11, wherein, during the cooling operation of the thermoelectric element, the second heating element heats the sample holder at a temperature lower than a temperature to heat the sample holder during the heating operation.
The thermal module according to claim 11, wherein the first thermal conductor and the second thermal conductor are thermally connected to different planes of the sample holder.
The thermal module according to claim 11, wherein the second heating element is a resistance heating module.
The thermal module according to claim 11, wherein the second thermal conductor comprises a metal layer.
The thermal module according to claim 1, further comprising a heat radiator emitting heat absorbed from the first thermal conductor during the cooling operation of the thermoelectric element.
The thermal module according to claim 17, wherein the heat radiator comprises:

a heat sink absorbing and emitting the absorbed heat generated by the cooling operation of the thermoelectric element; and

a ventilation fan cooling the heat sink.
The thermal module according to claim 18, wherein the heat radiator further comprises a metal block thermally connecting the thermoelectric element and the heat sink.
A method of operating a thermal module comprising a first thermal conductor to which a sample holder is to be thermally connected, the method comprising the operations of:

heating the sample holder by a first heating element thermally connected to the first thermal conductor; and

cooling the sample holder by a thermoelectric element thermally connected to the first thermal conductor.
The method according to claim 20, wherein, in the operation of heating, the thermoelectric element heats a plane thereof thermally connected to the first thermal conductor to the extent that the cooling operation after the heating operation is enhanced.
The method according to claim 20, wherein, in the operation of the cooling, the first heating element stops heating the sample holder.
The method according to claim 20, wherein the first thermal conductor includes two planes, and

the sample holder, the first heating element, and the thermoelectric element are thermally connected to the two planes.
The method according to claim 23, wherein the two planes include a top plane and a bottom plane.
The method according to claim 23, wherein the two planes include a front plane and a back plane.
The method according to claim 23, wherein the first heating element and the thermoelectric element are thermally connected to different planes of the first thermal conductor.
The method according to claim 20, wherein the thermal module further comprises a second thermal conductor thermally connected to the sample holder, and

in the operation of the heating, a second heating element thermally connected to the second thermal conductor heats the sample holder.
The method according to claim 27, wherein the first heating element and the second heating element heat the sample holder at the same time.
The method according to claim 27, wherein, in the operation of the cooling, the second heating element heats the sample holder at a temperature lower than a temperature to heat the sample holder in the operation of the heating.
The method according to claim 20, wherein the operation of the heating and the operation of the cooling are each repeatedly performed a plurality of times.