KR20230068624A - Apparatus for polymerase chain reaction - Google Patents

Abstract

A polymerase chain reaction device, which can perform a work for centrifugation of a sample and a work of replicating a genetic material with one device, comprises: a case with an open upper side; a lid opening and closing the open upper side of the case; a rotor rotatably disposed inside the case and equipped with at least one tube containing a sample; a rotor drive motor providing power to rotate the rotor; a heater in surface contact with the rotor to heat at least one tube mounted on the rotor; a cooling unit for cooling at least one tube heated by the heater; and a blower guiding cold air generated by the cooling unit to at least one tube and guiding hot air generated by the heater to the outside of the case. The heater and cooling unit heats and cools at least one tube according to a preset cycle to amplify a specific genetic material contained in the sample contained in at least one tube, and the rotor rotates at a high speed for centrifugation of the sample contained in at least one tube, or rotates at a low speed slower than the high speed to heat and cool at least one tube according to the preset cycle.

Description

Polymerase chain reaction apparatus {Apparatus for polymerase chain reaction}

The present invention relates to a polymerase chain reaction device, and more particularly, to a polymerase capable of centrifuging a sample and replicating a genetic material extracted by centrifugation through a polymerase chain reaction in one device. It is about a chain reaction device.

Polymerase chain reaction (PCR) is a molecular biology technique for enzymatic replication of nucleic acids without using living organisms. It is used for identification of fingerprints, diagnosis of infectious diseases, gene cloning, paternity testing, or DNA computing.

The polymerase chain reaction includes separating nucleic acids from a sample through centrifugation, adding a polymerase to the separated nucleic acids, and explosively amplifying nucleic acids by repeatedly heating and cooling at a specific temperature and time. Therefore, in order to carry out the polymerase chain reaction, a centrifugal separator for centrifuging a sample and a genetic material duplicator for duplicating the genetic material by heating and cooling the separated genetic material together with the polymerase are required. In a permanently installed institution such as a research institute or a general hospital, the polymerase chain reaction may be performed by separately providing the centrifugal separator and the genetic material replicator. However, it is difficult for temporary organizations or institutions such as simple infectious disease testing centers to separately prepare the centrifugal separator and genetic material replicator for reasons such as budget and space constraints and the need for rapid response, so that the urgent demand for infectious disease testing cannot be met. does not exist.

Registered Patent Publication No. 10-1433208

The present invention provides a polymerase chain reaction device capable of centrifuging a sample and replicating a genetic material extracted by centrifugation in one device.

In the present invention, a case with an open upper side, a lid that opens and closes the open upper side of the case, and at least one tube rotatably disposed inside the case and containing a sample are mounted. a rotor, a rotor driving motor for providing power to rotate the rotor, a heater in surface contact with the rotor to heat at least one tube mounted on the rotor, and the heater A cooling unit for cooling at least one heated tube, and guides cool air generated by the cooling unit to the at least one tube, and heat generated by the heater to the A blower leading to the outside of the case is provided, and the heater and cooling unit heats and cools the at least one tube according to a predetermined cycle to amplify a specific dielectric material contained in a sample contained in the at least one tube, , The rotor provides a polymerase chain reaction device that rotates at a high speed so that the sample contained in the at least one tube is centrifuged, or rotates at least one tube heated and cooled according to the preset cycle at a lower speed than the high speed. do.

The polymerase chain reaction apparatus of the present invention, in which the at least one tube is inserted and fixed one by one, may further include at least one tube holder detachably mounted on the outer circumference of the rotor.

Each of the at least one tube holder may be tilted with respect to the rotor.

The case has an inner partition wall separating the inside of the case into two inner spaces, and a partition wall opening is formed in the inner partition wall to allow air to flow between the two inner spaces. It is disposed in one of the two inner spaces, and when the cooling unit and the blower operate, cool air generated by the cooling unit moves from one of the two inner spaces to the other through the partition wall opening. By moving into space, the at least one tube may be cooled.

The rotor is rotatably disposed about an axis in a vertical direction, the heater is fixed to the rotor to rotate together with the rotor, and the blower is disposed under the rotor so that the blower operates inside the case. An air flow flowing from top to bottom along the axis and discharged to the outside of the case may be formed.

The polymerase chain reaction device of the present invention can perform centrifugation of a sample and replication of the centrifugally extracted genetic material with one device. Accordingly, since it is not necessary to separately prepare a centrifugal separator and a genetic material replicator, the initial set-up cost for performing the polymerase chain reaction is reduced. In addition, when the polymerase chain reaction device of the present invention is applied, a temporary organization or institution such as a simple infectious disease testing center can be quickly installed to respond to the urgent need for infectious disease testing.

1 is a perspective view of a polymerase chain reaction device according to an embodiment of the present invention.
Figure 2 is an exploded perspective view of the internal configuration of the polymerase chain reaction device of Figure 1;
FIG. 3 is a front view illustrating only a portion of FIG. 2 except for a cooling unit;

Hereinafter, a polymerase chain reaction apparatus according to an embodiment of the present invention will be described in detail with reference to the accompanying drawings. Terminologies used in this specification are terms used to appropriately express preferred embodiments of the present invention, which may vary according to the intention of a user or operator or conventions in the field to which the present invention belongs. Therefore, definitions of these terms will have to be made based on the content throughout this specification.

1 is a perspective view of a polymerase chain reaction device according to an embodiment of the present invention, FIG. 2 is an exploded perspective view of the internal configuration of the polymerase chain reaction device of FIG. 1, and FIG. 3 is only a part except for the cooling unit in FIG. It is a front view shown. 1 to 3 together, the polymerase chain reaction device 10 according to an embodiment of the present invention includes a case 11, first and second lids 22 and 23, a cooling unit ( 25), a rotor 31, a heater 45, a rotor driving motor 51, and a blower 61.

The case 11 includes a bottom plate 12, a side wall 14 surrounding an outer circumferential edge of the bottom plate 12 and extending upward from an outer circumferential edge of the bottom plate 12, and the bottom plate 12 An internal partition wall 17 is provided to divide the inside of the case 11 defined by the side wall 14 into two internal spaces 20 and 21 . One of the two internal spaces 20 and 21 is a PCR space 20 in which a centrifugal separation operation using the tube 2 containing the sample or a genetic material replication operation is performed, and the other one has a cooling unit 25 inside. It is a cooling unit space 21 installed in.

An open partition wall opening 18 is formed at an upper end of the internal partition wall 17 to allow air to flow between the PCR space 20 and the cooling unit space 21 . An air discharge through hole 15 and a heat dissipation through hole 16 are formed at the lower end of the side wall 14 . The air discharge through hole 15 is a through hole for discharging heat or cold air from the PCR space 20 to the outside of the case 11, and the heat dissipation through hole 16 is the heat generated from the condenser 28 of the cooling unit 25. It is a through hole for discharging to the outside of the case 11. The first lid 22 reliably closes the open upper side of the PCR space 20, and the second lid 23 reliably closes the open upper side of the cooling unit space 21.

The refrigeration unit 25 installed inside the refrigeration unit space 21 includes a compressor 27, a condenser 28, an expansion valve 29, and an evaporator 26. The compressor 27 compresses the refrigerant in a low-temperature, low-pressure gas state into a high-temperature, high-pressure gas. In the condenser 28, the high-temperature and high-pressure gaseous refrigerant passing through the compressor 27 releases heat to the surrounding air and becomes a high-temperature and high-pressure liquid state. In the expansion valve 29, the high-temperature and high-pressure liquid state refrigerant that has passed through the condenser 28 rapidly expands in volume and becomes a low-temperature and low-pressure liquid state refrigerant. In the evaporator 26, the low-temperature, low-pressure liquid refrigerant passing through the expansion valve 29 absorbs heat from the surrounding air to become a low-temperature, low-pressure gas, and the surrounding air becomes cold air. The low-temperature, low-pressure gaseous refrigerant that has passed through the evaporator 26 is introduced into the compressor 27 again.

Inside the cooling unit space 21, the condenser 28 and the evaporator 26 are spaced apart as much as possible. Preferably, the condenser 28 is disposed proximate to the heat dissipation through-hole 16 and the evaporator 26 is disposed proximate to the partition through-hole 18 .

Inside the PCR space 20, a rotor 31, a rotor driving motor 51, a heater 45, an intermediate separation member 55, and a blower 61 are disposed along the vertical axis AX. do. The rotor 31 is a wheel-shaped member disposed rotatably around the axis AX in the inside of the case 11, specifically, in the PCR space 20. A plurality of tubes 2 containing samples are mounted on the rotor 31 . The rotor 31 includes a cylindrical central pillar 32 disposed on the axis AX, an upper ring 35 surrounding the central pillar 32, a lower ring 39, and an intermediate ring ( 38) and a plurality of spokes 34 connecting the central pillar 32 and the upper ring 35. The upper ring 35 and the lower ring 39 are vertically spaced apart, and the middle ring 38 connects the inner circumferential edges of the upper ring 35 and the lower ring 39 .

A tube holder mounting portion 36 on which a plurality of tube holders 42 are detachably mounted is provided on the outer circumference of the upper ring 35 . The tube holder mounting portion 36 may be a dent that is dug inward from an outer circumferential edge of the upper ring 35 . The tube holder 42 is a member in the shape of a hexahedron, and a fitting through hole 43 is formed in the vertical direction so that the tube 2 is inserted therethrough. The tube 2 containing the sample may be inserted into and fixed to the fitting through hole 43 . The tube holder 42 mounted on the tube holder mounting part 36 may be tilted with respect to the rotor 31 . Accordingly, the inclination angle of the tube holder 42 and the tube 2 inserted into the tube holder 42 with respect to the axis line AX can be adjusted.

The rotor driving motor 51 includes a motor body 52 disposed below the rotor 31 and protruding upward from the motor body 52 along an axis line AX to the center of the rotor 31. A motor shaft 53 coupled to the pole 32 is provided. The rotor driving motor 51 provides power to rotate the rotor 31 about the axis AX.

The heater 45 is a ring-shaped member that is in surface contact with the rotor 31 to heat the plurality of tubes 2 mounted on the rotor 31, and a bolt and They are fixedly coupled by the same fastener. Specifically, the heater 45 has an intermediate ring 48 in surface contact with the inner circumferential surface of the intermediate ring 38 of the rotor 31, and a surface on the lower surface of the inner circumferential portion of the upper ring 35 of the rotor 31. The upper ring 47 protrudes from the upper edge of the intermediate ring 48 toward the axis line AX to be in contact with the lower edge of the intermediate ring 48 to be in surface contact with the lower surface of the lower ring 39 of the rotor. It has a lower ring 49 protruding in a radial direction from. The upper ring 47, the middle ring 48, and the lower ring 49 of the heater 45 have heating elements such as, for example, heating wires. The cooling unit 21 is mounted on the rotor 31 and cools the tube 2 heated by the heat generated by the heater 45 .

The blower 61 guides the cold air generated by the cooling unit 21 to the plurality of tubes 2 mounted on the rotor 31, and the hot air generated by the heater 45 is guided to the outside of the case 11. The blower 61 is disposed below the rotor 31 , the heater 45 , and the rotor drive motor 51 . The blower 61 may be a fan having a plurality of fan blades that pressurize air by rotating about an axis AX.

When the blower 61 operates, an air flow that flows from top to bottom along the axis AX in the PCR space 20 and is discharged to the outside of the case 11 through the air discharge hole 15 is formed. When the heater 45 operates to heat the plurality of tubes 2 mounted on the rotor 31, the blower 61 may not operate. This is because the heat from the heater 45 can be dissipated by the air flow induced by the blower 61 .

In order to cool the plurality of tubes 2 heated by the heater 45, the cooling unit 25 and the blower 61 may operate together. When the cooling unit 25 and the blower 61 operate together, the cool air generated by the cooling unit 25, specifically, the air cooled by heat exchange with the evaporator 26 passes through the bulkhead through hole 18. It moves from the cooling unit space 21 to the PCR space 20 through the rotor 31, cools the plurality of tubes 2, and descends to the case 11 through the air discharge through hole 15. released to the outside Therefore, the plurality of tubes 2 can be quickly cooled to a set temperature.

The intermediate separation member 55 divides the PCR space 20 into a lower space where the blower 61 is disposed and an upper space above it. The intermediate separation member 55 is a wheel-shaped member that includes a central pillar 56, an outer ring 59 disposed outside the central pillar 56, and the central pillar 56 and the outer ring 59. ) is provided with a plurality of spokes (spokes) 58 connecting them. The bottom plate 12 has a support protrusion 65 protruding upward along an axis line AX on an upper side thereof. The support protrusion 65 may support the motor body 52 of the rotor driving motor 51 by passing through the central pillar 56 of the blower 61 and the intermediate separation member 55 .

However, the polymerase chain reaction device of the present invention does not necessarily have to include the intermediate separation member 55, and the rotor driving motor 51 may be fixedly supported at the center of the blower 61. Although not shown, the polymerase chain reaction device of the present invention includes a temperature sensor for measuring the temperature around the tube 2 or the tube 2 itself mounted on the rotor 31, and the temperature measured by the temperature sensor. A controller for controlling operations of the heater 45 , the cooling unit 25 , the rotor drive motor 51 , and the blower 61 may be further provided.

The rotor 31 rotates at high speed so that the samples contained in the plurality of tubes 2 are centrifuged. The heater 45 and the cooling unit 25 use the plurality of tubes 2 mounted on the rotor 31 according to a predetermined cycle to amplify a specific genetic material included in the sample contained in the plurality of tubes 2 to heat and cool. In addition, the rotor 31 rotates the plurality of tubes 2 that are heated and cooled according to the preset cycle at a low speed lower than the high speed.

An example of a genetic material replication process using the polymerase chain reaction device 10 is as follows. Inject the sample containing the specific genetic material to be replicated, the primer, and the genetic material polymerase into the tube 2, close the cap 3 of the tube 2, and The tube 2 covered with the cap 3 is inserted into the tube holder 42, and the tube holder 42 is mounted on the tube holder mounting portion 36 of the rotor 31. Then, the rotor 31 is rotated at high speed to centrifugate the sample contained in the tube 2 to form a liquid layer containing a specific genetic material to be replicated. Then, according to a predetermined cycle, the tube 2 is heated to a temperature higher than 90 ° C, for example, 94 ° C, then cooled to a temperature of about 50 ° C, for example, 55 ° C, then to a temperature of about 70 ° C, for example, By repeating the process of heating again to 72 ° C., the specific genetic material is copied in large quantities. When heating and cooling the tube 2 according to a predetermined cycle, the rotor 31 may be rotated at a low speed to promote the polymerase chain reaction. Such mass-cloned genetic material can be identified through, for example, real-time PCR.

Meanwhile, only a sample containing a specific dielectric material is injected into the tube 2, the rotor 31 is centrifugally rotated to form a liquid layer including the specific dielectric material, and only the liquid layer is left in the centrifuged sample. , primer and dielectric material polymerase are injected into the tube 2 leaving only the liquid layer, the tube 2 is closed with the tube cap 3, and the tube 2 is mounted on the rotor 31 again. In addition, the specific dielectric material may be copied in large quantities by heating and cooling the tube 2 according to a predetermined cycle while rotating the rotor 31 at a low speed.

The polymerase chain reaction device 10 described above can perform centrifugation of a sample and replication of the genetic material extracted by centrifugation as one device. Accordingly, since it is not necessary to separately prepare a centrifugal separator and a genetic material replicator, the initial set-up cost for performing the polymerase chain reaction is reduced. In addition, when the polymerase chain reaction device 10 is applied, a temporary organization or institution such as a simple infectious disease testing center can be quickly installed to respond to urgent infectious disease testing needs.

Although the present invention has been described with reference to the embodiments shown in the drawings, this is only exemplary, and those skilled in the art will understand that various modifications and equivalent other embodiments are possible therefrom. Therefore, the true scope of protection of the present invention should be defined only by the appended claims.

2: tube 10: polymerase chain reaction device
11 case 25 cooling unit
31: rotor 45: heater
51: rotor drive motor 61: blower

Claims (5)

a case with an open top; a lid that closes the open upper side of the case to be openable; a rotor rotatably disposed inside the case and on which at least one tube containing a sample is mounted; a rotor drive motor providing power to rotate the rotor; a heater in surface contact with the rotor to heat at least one tube mounted on the rotor; a cooling unit for cooling at least one tube heated by the heater; and a blower for guiding cool air generated by the cooling unit to the at least one tube and guiding hot air generated by the heater to the outside of the case,
The heater and cooling unit heats and cools the at least one tube according to a predetermined cycle to amplify a specific genetic material included in the sample contained in the at least one tube,
The rotor rotates at a high speed so that the sample contained in the at least one tube is centrifuged, or rotates at least one tube heated and cooled according to the preset cycle at a lower speed than the high speed. Polymerase chain reaction Device. According to claim 1,
The polymerase chain reaction device further comprising: at least one tube holder in which the at least one tube is inserted and fixed one by one and is detachably mounted on the outer circumference of the rotor. According to claim 2,
Polymerase chain reaction device, characterized in that the at least one tube holder is tiltable with respect to the rotor, respectively. According to claim 1,
The case has an inner partition wall separating the inside of the case into two inner spaces,
A partition wall opening is formed in the inner partition wall to allow air to flow between the two inner spaces,
The cooling unit is disposed in one of the two internal spaces,
When the cooling unit and the blower operate, the cool air generated by the cooling unit moves from one of the two inner spaces to the other one of the two inner spaces through the partition wall opening, thereby cooling the at least one tube. Polymerase chain reaction device characterized by. According to claim 4,
The rotor is rotatably disposed about an axis in the vertical direction,
The heater is fixed to the rotor to rotate together with the rotor,
The blower is disposed below the rotor, and when the blower operates, the polymerase chain reaction device flows from top to bottom along the axis inside the case and an air flow discharged to the outside of the case is formed. .

Images