KR100855996B1 - PCR disc apparatus, PCR disc driver apparatus, and analysis method using the same - Google Patents

Abstract

The present invention relates to a PCR disk device, a PCR disk driver device, and an analysis method using the same. More particularly, the present invention relates to amplification of DNA or RNA necessary for various diagnostic analysis devices, nucleic acid hybrid analysis devices, or immunological verification devices. The present invention relates to a PCR disk in which a PCR process is batch-arranged, a controller for controlling the PCR disk, and a device for a PCR disk driver for driving and controlling the PCR disk, and an analysis method using them. PCR disk apparatus according to the present invention comprises a solid substrate comprising a flow path for the fluid flow, a chamber for storing the buffer solution and a hole connecting the flow path; And PCR disk including a valve for opening and closing the hole; A control disc including a control unit for supplying a power or control signal necessary for the PCR disc; And an interface unit for connecting the PCR disk and the control disk.

Electromagnet, Memory, Brush, Air Vent

Description

PCR disc apparatus, PCR disc driver apparatus and analysis method using the same {PCR disc apparatus, PCR disc driver apparatus, and analysis method using the same}

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1A and 1B are sectional views showing a valve device using cylindrical micro particles installed in a PCR disc 100 according to an embodiment of the present invention.
FIG. 1C illustrates an embodiment of a PCR disk in which a PCR process is batch arranged and a control disk 200 for controlling the PCR disk.
1D is a cross-sectional view showing the structure of a conductor arm according to an embodiment of the present invention.
1E is a cross-sectional view showing an embodiment of a PCR disc driver device in which a control disc 200 is incorporated.
2A is a cross-sectional view illustrating one embodiment of an integrated PCR disk 700.
2B is a cross-sectional view illustrating one embodiment of a PCR disk driver for the integrated PCR disk 700.
Figure 3a shows an embodiment of a PCR disk including the preparation process and the PCR process.
Figure 3b shows an embodiment of an electrophoretic disk including electrophoresis.
3C shows a cross-sectional view of a PCR disc for a chamber according to the main process of FIG. 3A.
Figure 3d shows an embodiment of a heater and a magnetic field generating device required for the preparation process and the PCR process.
3E illustrates an embodiment for implementing a strong electromagnet using low current.
4A and 4B show an embodiment of the appearance of the PCR disc driver device, respectively.
FIG. 4C illustrates an embodiment of a PCR disc detection method for determining whether a currently loaded disc is an optical disc or a PCR disc such as a general music CD, CD-R, game CD, or DVD. An example is shown.
<Explanation of symbols for main parts of drawing>

100: PCR disk 108109: brush

63 control part 70a valve

200: control disk 4a: electromagnet

188: memory

The present invention relates to a PCR disk device, a PCR disk driver device, and an analysis method using the same. More particularly, the present invention relates to amplification of DNA or RNA necessary for various diagnostic analysis devices, nucleic acid hybrid analysis devices, or immunological verification devices. The present invention relates to a PCR disk in which a PCR process is batch-arranged, a controller for controlling the PCR disk, and a device for a PCR disk driver for driving and controlling the PCR disk, and an analysis method using them.

Until recently, most clinical diagnostic analytical devices for detecting small amounts of analytes in fluids have been designed for efficiency and economics by designing devices for multiple sample preparation and automated reagent addition and for analyzing large numbers of test samples in parallel or in series. Improvements were made. Often these automated reagent preparation devices and automated multiple analyzers are integrated into a single device. This type of clinical analyzer can accurately and automatically perform hundreds of analyzes with a small sample and reagents within an hour.

However, these analyzers are expensive and only centralized laboratories and hospitals can purchase them.

Therefore, in order to overcome such a problem, there is an urgent need for a small, automated PCR device that can be easily handled by anyone.

<Disk>

A standard compact disc is formed from a 12 cm polycarbonate substrate, reflective metal layer and protective lacquer coating. The format of CDs and CD-ROMs is described by the ISO 9660 industry standard.

The polycarbonate substrate is an optical quality transparent polycarbonate. In standard printed or mass replicated CDs, the data layer is part of a polycarbonate substrate and the data is engraved in a series of pits by a stamper during the injection molding process. During this injection molding process, molten polycarbonate is injected into the mold under high pressure and then cooled so that the polycarbonate has a mirror image of a mold or "stamp" or "stamp" and a pit representing binary data on the disk substrate is produced and mastered. It is held by the polycarbonate substrate as a mirror image of the pit of the stamper generated during the mastering process. The stamping master is usually glass.

The polycarbonate art substrate may be modified and modified by a thin film type analyzer for diagnosing and detecting a small amount of a substance in a fluid such as a PCR disc. A chamber may be formed for storing channels and buffer solutions through which flow can flow. There is a need for a microvalve capable of smoothly controlling the flow and flow rate of the fluid in the flow path formed in such a thin film and electronic control for controlling them.

In addition, in the present invention, a chamber may be formed in a silicon wafer to store a channel and a buffer solution through which a fluid may flow by a semiconductor process. Also provided is a PCR disk in which electronic circuits for smoothly controlling the flow and flow of fluid in a flow path formed in such a thin film are integrated on a silicon wafer.

As a generally known technique, an analysis apparatus using an optical compact disc is known as "confocal compact scanning optical microscope based on compact disc technology" (1991, Vol. 30, No, 10, Applied Optics), "Gradient-index pobjectives for CD applications" ( April, 1987, Vol26, Issue 7, Applied Optics).

In addition, as a device for forming a fluid film on the surface of the disk using a centrifugal force, the sample injected into the inlet on the disc, the European patent "Disc for centrifuge" (GB1075800, publication date 1967.07.12) and the fluid injected into the inlet on the disc European Patent "Separating Disks for centrifuge" (3,335,946, April 12,1965) is disclosed as a device for separating a sample while moving the sample to a channel and a chamber using centrifugal force.

Also disclosed is a US patent "disc centrifuge photosedimentometer" (4,311,039, Jan. 19,1982) as an apparatus for chemical analysis by centrifugal force and optical measurement on a disc.

However, this has the disadvantage of not being able to fully automate the process required for PCR, which is not suitable for automated PCR process construction.

The present invention has been made to solve the problems of the prior art, an object of the present invention is to provide DNA from a sample such as blood to amplify DNA or RNA, which is essential in various diagnostic analysis devices, nucleic acid hybrid analysis devices and immunological verification devices, etc. Another object of the present invention is to provide a PCR disk device capable of automatically performing an entire PCR process for amplifying RNA.

Another object of the present invention is to provide an integrated PCR disk apparatus having a configuration in which the PCR disk of the present invention and a control disk capable of controlling all of the processes thereof are integrated into one disk.

Still another object of the present invention is to provide a PCR disk device capable of performing not only a PCR process but also an electrophoretic process.

Still another object of the present invention is to provide a device for a PCR disk driver, which includes a control unit for controlling the PCR disk of the present invention and controls the driving of the PCR disk.

Still another object of the present invention is to provide an analysis method using the PCR disk device or PCR disk driver device of the present invention.

Hereinafter, with reference to the accompanying drawings will be described in detail a preferred embodiment of the present invention.

The valve for controlling the flow or flow of the fluid required for the PCR process installed on the PCR disk of the present invention is provided with a micro bead (Mini bead) moving inside the PCR disk by the magnetic force formed by the electromagnets installed on the upper and lower sides of the PCR disk To open and close the flow path.

In a preferred embodiment of the present invention, the microbeads include materials such as, for example, magnet balls, ferromagnetic metal particles, phase magnetic particles, semimagnetic particles, metal balls, and the like. In addition, the micro beads may be formed of solid metal, plastic, glass beads, or the like, and a metal coating may be formed thereon. The metal sphere may also be an alloy. In addition, the micro-bead can be charged, in this case using an electrode plate disposed on the upper and lower surfaces of the PCR disk instead of the electromagnet. Charged beads open or close the pores according to the direction of the voltage applied to the electrode plate. The microbeads have a diameter in the range of 1 μm to 1 mm, preferably 100 μm to 500 μm. As the size of the beads increases, the contact surface increases, thereby increasing the reliability of opening and closing. The electromagnet may be an air core or a non-magnetic core. The diameter of the winding is preferably 0.01mm ~ 0.1mm. More preferably, the micro beads may be ultra-small magnet particles having a cylindrical shape of 0.1 mm to 0.5 mm thickness. The diameter of the cylindrical magnet particles is preferably 0.1 mm to 5 mm.

1A and 1B are cross-sectional views illustrating a valve device using cylindrical micro particles installed in a PCR disc 100 according to an embodiment of the present invention.

The PCR disc 100 is composed of an upper substrate 1, an intermediate substrate 2, and a lower substrate 3.

FIG. 1A illustrates a case in which the oil holes 10b are blocked by the cylindrical magnet particles 70 and the flow paths 22a and 22c are blocked, and FIG. 1B shows that the oil holes 10b are opened to connect the flow paths 22a and 22c. As shown in FIG. 1A, to close the oil hole 10b to block the flow path, the power is cut off to the upper electromagnet 4a and the power is applied to the lower electromagnet 5a facing the oil hole 10b. To connect the flow path by opening the hole 10b as shown in FIG. 1B, power is applied to the upper electromagnet 4a and the power is cut off to the lower electromagnet 5a. Alternatively, opening and closing of the valve may be controlled by controlling an out-of-winding current direction of the electromagnets 4a and 5a so as to cause a repulsive force or suction force with respect to the cylindrical magnet particles 70.

The electromagnet of the present invention (4a, 5a) is preferably an electromagnet of the air core (air core), the micro-bead is preferably a magnet ball (magnet ball) or cylindrical micro-shaped magnet particles.

In addition, in the present invention, since the narrow channels 22a and 22c are formed in the disk for fluid movement, the exhaust port 12 is connected to the upper substrate 1 so that the fluid flows smoothly without pressure. Formed.

In addition, when the flow path is connected as shown in Figure 1b, there is a restriction groove 101 so that the ultra-small cylindrical magnet particles 70 are in close contact with the upper substrate (1). The restraint groove 101 is to prevent the magnet particles 70 from falling out by the disk shaking to block the open hole 10b.

FIG. 1C illustrates an embodiment of a PCR disk in which a PCR process is batch arranged and a control disk 200 for controlling the PCR disk.

More specifically, FIG. 1C is a flow chart for storing various buffer solutions required for analysis, a chamber for performing various chemical processes, a flow path for moving the treated fluid and buffer solutions, and controlling the opening and closing of these flow paths. One embodiment of a PCR risk 100 in which a PCR process including valve devices is disposed and a control disc 200 for controlling the PCR risk 100 are provided.

The PCR disc 100 of the present invention can be selected from a variety of materials, such as plastic, PMMA, glass, mica, silica, silicon wafer materials, and the like. However, plastics are preferred for economic reasons and for ease of processing. Plastics that can be used include polypropylene, polyacrylates, polyvinyl alcohols, polyethylene, polymethylmethacrylates, and polycarbonates. Polypropylene and polycarbonate are preferred, with polycarbonate being most preferred.

The PCR disc 100 consists of an upper substrate 1, an intermediate substrate 2 and a lower substrate 3, each of which is a channel and a buffer through which fluid can flow on the substrate surface during the injection molding process. A plurality of holes for connecting the passage and a chamber for storing a buffer solution are formed.

These are closely attached to each other to form a single PCR disk 100 body.

The control disk 200 may be selected from a variety of materials, such as plastic, PMMA, glass, mica, silica, silicon, the material of the printed circuit board (PCB).

However, in the present invention, the material of the PCB material and the silicon wafer is preferable because of the ease of circuit design.

The PCR disc 100 is formed by laminating an upper substrate 1, an intermediate substrate 2, and a lower substrate 3. The ultra-small cylindrical magnet particles 70a and 70b are independently opened and closed by magnetic force formed by the electromagnets 4a and 5a and 4b and 5b, respectively. Reference numeral 120 denotes a pipette or syringe for injecting a sample, 121 denotes a sample inlet, 170 denotes a disk gap, 171b denotes an interface for connecting the control disc 200 and 171a, and a control signal to the PCR disc 100. A plurality of control signal connection unit 173b for supplying and a power supply connection 31b for supplying power. The surfaces of the connecting portions 173b and 31b are composed of connecting grooves 33 coated with a conductor surface or a conductor.

Reference numeral 131 is a preparation chamber including a preparation process for preparing DNA samples from blood or cells by RT (Reverse Transcription) from RNA, and 132 is a PCR (Polymerase Chain). PCR chamber for reaction process.

Reference numerals 130 and 140 denote chambers containing buffer solutions containing various enzymes necessary for the constant preparation process.

Reference numerals 141 and 142 denote chambers containing buffer solutions containing various enzymes including polymerases and primers required for the PCR process.

The opening and closing control of the valve at the start and end of each process (prep process, PCR process) is performed by the power applied to the electromagnets 4a and 5a, 4b and 5b provided on the upper and lower sides of the ultra-small cylindrical magnet particles. It is made by on-off or current direction control, and the fluid movement is by centrifugal force by the rotational force of the disk.

In addition, power is supplied to the control unit 63 and the transmission / reception apparatus 107 on the control disk 200 by frictional contact or wireless transmission (non-contact interface) between the control disk 200 and the brushes 108 and 109.

An annular electrode plate 223 for friction contact between the brushes 108 and 109 may be installed on the upper or lower plate of the control disk 200. In addition, the control signal and the power of the controller 63 are connected to the conductor arm 13 by a pattern of a circuit with the connection parts 173a and 31a in the region 171a of the control disc 200.

Power supply and control signal transmission from the control disk 200 to the PCR disk 100 are connected to a plurality of conductor arms 13 connected to the connection portions 173b and 31b of the interface region 171b on the PCR disk 100. Made by them. These are made in the form of a conductive arm 13 and the connecting groove 33, and serves as a connector between the PCR disk 100 and the control disk 200. The conductor arm 13 is rotated at the same time by engaging the connecting groove 33 by the pressing contact between the control disk 200 and the PCR disk 100 at the start of the rotation or operation of the disk.

The power supplied by the brushes 108 and 109 is supplied through a power connection 31a on the control disk 200 and a power connection 31b on the conductor arm 13 and the PCR disk 100. The control signal supplied through 173a is connected by a plurality of connection grooves 173b.

In addition, the connecting grooves 33 on the PCR disk 100 connected to the conductor arm 13 are connected to the circuit pattern again to supply various control signals and power necessary for each section of the PCR disk 100.

Reference numeral 188 denotes a memory containing information related to a protocol for an PCR process, an analysis algorithm, a standard control value for reading, and bioinformatics information. It can also include device driver software for PCR disk drivers and patient education information for clinical analysis and can be modified. It can also include web sites and links to connect with specialists and hospitals based on the results of the diagnosis. In addition, personal encryption information can be stored so that others can not be used without permission.

The memory 188 is preferably RAM or ROM. The memory 188 information is provided to the central control unit in the PCR disc driver via the conductor arm 13 and the contactless interface 107 and is utilized for personal encryption.

1D is a sectional view of an example showing the structure of the conductor arm 13.

During the rotation of the disk, the interface of the control disk 200 so that the conductor arm 13 can be smoothly inserted into the connecting groove 33 during the compression close contact between the control disk 200 and the PCR disk 100. A spring 13c is connected to one end of the conductor arm 13 which is fixedly connected to the portion 171a. Reference numerals 13a and 13b constitute a conductor arm, and the ends of 13b are inserted into the connecting groove 33. The spring 13c and the conductor arms 13a and 13b are interconnected by 13d so that the conductor arms are elastic.

FIG. 1E illustrates an embodiment of a PCR disc driver device in which the control disc 200 is mounted to control drive the PCR disc 100.

Brushes (109, 108) for supplying power supply to the control unit (63) on the rotating control disk (200); A power supply device 110 for supplying power thereto; 300 is the body supporting the PCR disk driver. A circuit board 145 is jointly fastened to the PCR disk drive body 300 on the bottom of the PCR disk drive, and a central control unit 101 and a storage device 111 for controlling the PCR disk driver on the circuit board are provided in the circuit. It is designed on the substrate 145. The central control unit 101 rotates and stops the motor 102 when the disks 100 and 200 rotate or stop.

In addition, the central control unit 101 transmits various control command signals necessary for controlling the PCR process through the non-contact interfaces 106 and 107 located in the control disk 200.

The control disk 200 receiving the control command supplies various control signals for controlling the PCR disk 100 through the conductor arm 13.

In addition, the control unit 63 displays the process progress and analysis results of the PCR disc through the infrared interface, optical interface, or wireless interface, which are the non-contact interfaces 106 and 107. Send to).

106 and 107 are receivers or transmitters or transceivers for the contactless interface, respectively. In addition, the central control apparatus 101 may transmit a control command to the control unit 63 through the non-contact interface to the control disk 200.

The contact interfaces 106 and 107 are made by an infrared sensor in the case of an infrared interface, and are made by a photo sensor in the case of an optical interface. The air interface is also made by a stripline antenna and a demodulation device therefor.

Reference numeral 188 denotes a memory containing information related to a protocol for an PCR process, an analysis algorithm, a standard control value for reading, and bioinformatics information. It can also contain patient training information for device driver soft control and clinical analysis and can be modified. In addition, it may include a web site and a variety of links that can be linked to the diagnosis results and doctors and hospitals according to the results. In addition, personal encryption information may be stored so that others cannot use it without permission.

The memory 188 is preferably RAM or ROM. The memory 188 information is provided to the central control unit 101 via a contactless interface with the conductor arm 13 and utilized for personal encryption.

The control disk 200 and the PCR disk 100 are seated in the PCR disk driver by the fastening portions 105a and 105b.

103 is a measuring device for measuring the final result of the PCR process.

2A to 2B show a case where the control disk 200 and the PCR disk 100 are combined to constitute one integrated PCR disk 700.

In this case, the integrated PCR disk 700 is preferably a silicon wafer material.

The silicon wafer material is a monolithic integrated circuit integrated with an electronic circuit including the controller 63, the memory 188, the non-contact interface 107, the electrode plate or electromagnet for controlling the valve, and various circuit patterns. Circuit) design is easy to configure, and the chamber and channel formation can be easily implemented by photolithography and etching processes.

Integrated circuit design, photolithography and general etching processes on wafers by these semiconductor processes are well known in the art.

The integrated PCR disk 700 is formed of the upper substrate (4), the intermediate substrate (5) and the lower substrate (6); The photolithography and various etching processes form a hole connecting the channel with the chambers 130, 131, 132, 140, 141 and 142 capable of storing the channel and the buffer solution through which the fluid can flow. Brushes 109 and 108 for supplying power supply and annular electrode plates 223 for frictional contact of the brushes 108 and 109 are provided on the bottom surface of the integrated PCR disc 700a. 170 is a disk void. 171a is an interface unit, and the control unit 63 of 700a includes a plurality of control signal connection units 173a for supplying control signals to valves and electronic circuits on 700b and a power connection unit 31a for power supply. In the above embodiment, the connection of the connection portions 173a, 173b and 31a, 31b is connected by vias or through holes in the interface portion 171a, but is not required to be limited to the interface portion 171a and vias at necessary positions on the disk. I can be connected by a through hole. Via or through hole design is well known to those skilled in the art.

In addition, the memory 188 may have an opening for erasing the contents of the ROM by ultraviolet (UV) when the ultraviolet (Ultra Violet) erasable ROM is used.

2B illustrates an embodiment of a PCR disc driver device for controlling the integrated PCR disc 700. The control unit 63 sends the process progress to the external central control unit 101 or the storage unit 111 through an infrared interface, an optical interface, or a wireless interface, which are non-contact interfaces 106 and 107. 106 and 107 are receivers or transmitters or transceivers for the contactless interface, respectively. In addition, the central control apparatus 101 may transmit a control command to the controller 63 to the integrated PCR disk 700 through the non-contact interface.

The non-contact interfaces 106 and 107 are made by an infrared sensor in the case of an infrared interface, and are made by a photo sensor in the case of an optical interface. The air interface is also made by a stripline antenna and a demodulation device.

A brush (109,108) for supplying power to the control unit (63) on the integrated PCR disk (700) which is rotating and a power supply (110) for supplying power to the brush (109,108). 300 is the body supporting the PCR disk driver. A circuit board 145 is jointly fastened to the drive body 300 at the bottom of the PCR disk drive, and a central control unit 101 and a storage device 111 for controlling the PCR disk driver are mounted on the circuit board. 145). The central control unit 101 not only stops the motor 102 when the integrated PCR disk 700 rotates or stops, but also controls the measuring device 103 for measuring the PCR result.

In addition, the central control unit 101 transmits various control command signals necessary for controlling the PCR process through the contactless interfaces 106 and 107 located in the integrated PCR disk 700.

Reference numeral 188 denotes a memory containing information related to a protocol for an PCR process, an analysis algorithm, a standard control value for reading, and bioinformatics information. It can also contain device driver software and patient education information for clinical analysis and can be modified. It can also include web sites and links to connect with specialists and hospitals based on the results of the diagnosis. In addition, personal encryption information may be stored so that others cannot use it without permission.

The memory 188 is preferably RAM or ROM. The memory 188 information is provided to the central control unit 101 via the contactless interfaces 106 and 107 and is utilized for personal encryption.

The integrated PCR disk 700 is seated in the PCR disk driver by the fastening portions 105a and 105b.

Figure 3a shows an embodiment of a PCR disk including the preparation process and the PCR process.

Reference numeral 170 denotes a void of the disk, and 171b provides an interface for connecting to the control disk 200.

Reference numeral 131 denotes a preparation chamber including a preparation process for preparing DNA samples from blood, cells, or RNA, and 132 is a PCR chamber for a polymer chain reaction (PCR) process. A chamber containing a buffer solution containing various enzymes including enzymes, primers, and the like, and 88a is an out chamber of a PCR product.

Figure 3a is an embodiment for the case of configuring a plurality of the PCR chamber.

Each of the plurality of PCR chambers 131a may contain different primers or the same primers. Alternatively, a plurality of primers may enter each of the plurality of PCR chambers.

DNAs amplified from each of the plurality of PCR chambers are combined in chamber 88a.

Reference numeral 130 is a chamber containing a washing buffer, 140 is a chamber containing distilled water. 144a is a chamber for storing the ladder marker, and 141a is a chamber containing positive or negative control. There are output chambers 88b and 88c for each of these.

142a and 143a are chambers containing various solutions required for electrophoresis, such as staining. There are output chambers 88d and 88e for each of these.

Reference numerals 150,151,152,153,154a, 155a, 156a, 157a denote valves.

The prep chamber 131 includes an electromagnet 177 and a heater.

The output chamber 88a may have an outlet for allowing conventional electrophoresis by manual operation by collecting a PCR product by a pipette or a syringe.

The electrophoresis is a known technique of the person in charge.

Figure 3b shows an embodiment of an electrophoretic disk including electrophoresis.

The electrophoresis disk is combined with the PCR disk to provide a PCR disk device capable of automated electrophoresis.

The electrophoresis disk is located on the bottom of the PCR disk.

The electrophoresis disk is provided with electrodes 555a and 555b for applying a voltage to a gel plate 555c consisting of agarosdegel and / or polyacrylamide gel. It is ..

The output chambers 88a, 88b, 88c, 88d, and 88e of the PCR product, ladder marker, positive / negative controls and stains are combined with the input chambers 99a, 99b, 99c, 99d, and 99e of the electrophoresis discs. In a one-to-one connection, when the valves 152a, 157a, 154a, 155a, 156a are opened, the contents of the output chamber flow into the input chamber of the electrophoretic disk and a voltage is applied to the electrodes. This is going on.

The applied voltage is removed to stop electrophoresis after a certain time.

The voltage control and valve control are by the control unit 63.

The control disk 200 is connected between the PCR disk and the electrophoretic disk by the interface unit (171b, 171c).

In addition, the PCR disk driver device having the electrophoretic disk includes an optical measuring device 103 including a UV (ultra violet) device and a camera to measure, store, and transmit the band generated on the gel plate 555c. In addition to being possible, it is possible to display on a computer monitor.

FIG. 3C is a cross-sectional view of the PCR disc for the chamber according to the main process of FIG. 3A.

The ultra-small cylindrical magnet particles 150, 151, 153 and 156 for the valve of the main process are independently opened and closed controlled by the magnetic force formed by the electromagnets 4a and 5a, 4b and 5b, 4c and 5c, respectively. 121 is a sample inlet.

Reference numeral 131 denotes a preparation chamber including a preparation process for preparing DNA samples from blood, cells, or RNA. The prep chamber contains ferromagnetic beads and cells with a Lysis buffer solution that breaks cells with affinity for DNA for DNA extraction.

132 is a PCR chamber for PCR (Polymer Chain Reaction) process, a chamber that stores a buffer solution containing various enzymes such as polymerase, dNTP, primer, etc. necessary for PCR process, and 88a is a PCR product. To store it.

3A-3C, one embodiment of the main process for the PCR disc is as follows.

<Prep process>

The preparation chamber 131 is a chamber for extracting DNA from a sample, and an embodiment of the preparation process is as follows.

1) 10 μl of blood (EDTA, ACD Tube) or 5 μl (Heparin Tube) is injected through the sample inlet 121 installed in the preparation chamber 131. The prep chamber 131 contains a Lysis buffer solution for breaking DNA and extracting DNA, and a ferromagnetic bead having affinity with the extracted DNA.

2) After incubation for 5 minutes, DNA is extracted from the cell and adhered to the ferromagnetic beads having affinity with DNA.

3) The electromagnet 177 of FIGS. 3A and 3B is turned on to allow the ferromagnetic beads to stand and then wait for 1 to 3 minutes.

4) While slowly rotating the PCR disc, the valve 161 is opened to wash debris from the cell destruction into the trech chamber 135. Close valve 161 and stop PCR disc rotation.

5) The electromagnet 177 is turned off.

6) While opening the valve 150 and slowly rotating the PCR disk, the washing buffer of the chamber 130 flows into the preparation chamber 131.

7) After repeating the process 3) to 6) twice, finally, while slowly rotating the PCR disc, the valve 161 is opened and washed with the trech chamber 135. Close valve 161 and stop PCR disc rotation.

8) After the electromagnet 177 is turned off, the valve 151 is opened while slowly rotating the PCR disc to introduce distilled water from the chamber 140 into the preparation chamber 131.

9) The DNA attached to the ferromagnetic beads may be separated by reheating using a heater 960 embedded in the preparation chamber 131 or resuspension using a resuspension buffer.

<PCR process>

The PCR chamber 132 is a chamber for amplifying DNA.

1) While slowly rotating the PCR disc, the valve 152 is opened to move DNA separated from the ferromagnetic beads in the preparation chamber 131 to the PCR chamber 132 by the preparation process.

2) After completion of DNA transfer to the PCR chamber, the valve 152 is closed and the PCR disc is stopped.

3) Amplify DNA by repeating the PCR cycle about 30 times using the heater 961 and the temperature sensor 520 embedded in the PCR chamber 132.

4) Cool it by rotating it for 1-2 minutes.

Thereafter, the DNA amplified in the plurality of PCR chambers 132 is slowly moved to the chamber 88a by opening the valve 153 while slowly rotating the disk.

Figure 3d is an embodiment of a heater and a magnetic field generating device required for the preparation process, PCR process. Reference numerals 960,961 denote heaters composed of electrode patterns or coil windings. Preferred coil windings are circular windings of air cores.

The electrode pattern 960 alone acts as a heater but also serves as the electromagnet 177 above to generate a magnetic field.

The temperature control and the magnetic field control are controlled by the controller 63. A large voltage is applied to the input terminals 961a and 961b so that a large current flows in the electrode patterns 960 and 961 so that the electrode pattern 962 acts as a resistance line or a heating line.

The controller 63 adjusts the temperature by supplying a voltage to the input terminals 961a and 961b of the electrode patterns 960 and 961 and measuring the temperature generated by the current electrode patterns 960 and 961 by the temperature sensor 520 to control the amount of current. Do it. In addition, the electrode pattern 960 serves as an electromagnet by flowing a low current.

3E illustrates an embodiment for implementing a strong electromagnet despite the use of a low current in the electrode pattern 960. This is important because strong magnetic force is required to fix the ferromagnetic beads in the preparation process (3).

Reference numeral 970 denotes a magnet made of a ferromagnetic material, and strongly adheres to the preparation chamber 131 even by a weak magnetic force by the electrode pattern 960, thereby generating a strong magnetic force in the preparation chamber. In addition, when the electrode pattern 960 is turned off or after generating a magnetic force corresponding to a temporary repulsive force, the magnet 970 is separated from the preparation chamber. Therefore, no magnetic force is present in the preparation chamber.

The temperature sensor 520 in the above embodiment was used only for a PCR process requiring precise temperature control, but may be used for all processes requiring temperature sensing.

The analysis starts automatically when the PCR disc is loaded into the PCR disc drive.

However, when the sample is loaded without being injected into the sample inlet, eject and / or a warning message is sent back to the user.

The sample injection is possible by further comprising an impedance measuring device in the preparation chamber (131). That is, when the sample is injected or not, the impedance characteristics are different, so whether the sample is injected can be known.

3E illustrates a sample injection based on an impedance change by an interdigitated array.

One embodiment of an impedance measuring device implemented by interdigitated array electrodes 702 and 703 is shown. As the number of digits of interdigitated array electrodes 702 and 703 increases, the sensitivity of the detector increases.

An AC signal having a certain bandwidth from the electronic controller 63 is applied to the two terminals 704 and 705 of the interdigitated array, thereby measuring the frequency response characteristic, thereby making it possible to measure impedance. Or simply measure the resistance.

If during the diagnosis or analysis of the sample by the PCR disc, when the user requests eject (un-loading) or stop of the PCR disc from the PCR disc driver, the PCR disc driver ignores the PCR disc driver and continues the analysis and diagnosis. At this time, a warning message is notified to the user or a password is required.

If the password is correct, the user accepts the eject (un-loading) or stop request.

Upon completion of diagnostics and analysis, it accepts the user's eject request, allowing the PCR disk to be ejected from the PCR disk driver.

In addition, the memory 188 stores information on how many times the disk has been used, information on expiration date, and the type of disease to be diagnosed.

That is, when the disposable PCR disc is used or ejected upon completion, a history of it is recorded in the memory 188, and the user is informed of the possibility of use when reloading later.

It also informs the user that the PCR discs that have expired cannot be amplified.

4A and 4B illustrate an appearance of a PCR disc driver device, where 751 is a case of the PCR disc driver device and 750 is a tray for loading the PCR discs 100 and 700. Further, the PCR disc driver device of the present invention has a play and seek button 745 and a stop button 746 for reproducing a conventional optical disc.

4A illustrates an embodiment in which a progress state of a PCR disc driver device is displayed using a light emitting diode. The PCR disk detecting means includes a PCR disk light emitting diode 741 which determines whether the PCR disk is currently loaded, and indicates that the currently loaded disk is a PCR disk, and a progress display light emitting diode 742 which displays the current analysis progress. Reference numeral 751 denotes a light emitting diode for indicating that a conventional optical disk has been loaded.

Therefore, the PCR disc driver device of the present invention is compatible with optical disc devices such as CD-ROM and DVD readers as well as driving the above-described PCR discs. It also has the ability to read and play discs, providing not only price competition but also convenience in use. In addition, the PCR, disk driver device is connected to a computer to facilitate remote diagnosis using the existing Internet network.

Figure 4b shows an embodiment of displaying the progress of the PCR disk driver apparatus using the liquid crystal display device.

Progress according to the main process (prep process, PCR process) can be displayed in percent (%) or bar graph (bar graph) format.

Alternatively, you can use a computer monitor to display the progress of the PCR disk driver device. Progress according to the main process (prep process, PCR process, electrophoresis process) can be displayed in the form of percent (%), bar graph (pie graph), pie graph (pie graph). In addition, the result of PCR measurement by the measuring device 103 can be displayed on the monitor.

FIG. 4C shows a PCR disc for detecting whether the currently loaded disc is an optical disc such as a music CD, a CD-R, a game CD, a DVD, or a PCR disc. Disc Detection) is an embodiment of the method.

The control disk 200 uses the conductor arm 13 and the connecting groove 33 to determine a constant voltage or a divided voltage (referred to as a bio-disc detection signal) generated by resistors R1 and R2 disposed on a PCR disk. This is done by the control unit 63 of. Since the resistors R1 and R2 are not disposed in a normal optical disk, the above constant voltage does not occur. Thereafter, the control unit 63 transmits the PCR-disc detection signal to the external central control unit 101 by wireless transmission through the non-contact interfaces 106 and 107 so that the currently loaded disc is the PCR disc. I can inform you.

In the case of a normal optical disc, the controller operates in a normal manner, and in the case of a PCR disc, the controller 63 sends various control signals for the PCR process to the PCR disc 100.

Although the present invention has been described in terms of embodiments and examples, it is not intended to limit the invention thereto. In addition, those skilled in the art will recognize that various modifications and variations are possible in addition to those described herein. Such modifications also fall within the scope of the appended claims.

As described above, the present invention relates to an apparatus and method for a PCR disk and a PCR disk driver using a disk. A PCR disk in which the process is batch placed; Provided is an apparatus and method of a PCR disk driver having a control unit for controlling the PCR to drive control the PCR disk.

In particular, the PCR disc driver device and method are compatible with optical disc devices such as CD-ROM and DVD readers as well as driving the PCR disc, so that the conventional optical CD, CD-R, game CD, DVD, etc. It also has the ability to read and play discs, providing not only price competition but also convenience in use. In addition, the PCR disk driver device is connected to a computer to facilitate remote diagnosis using an existing Internet network.

In another aspect, the present invention provides an integrated PCR disk apparatus and method having a configuration in which the PCR disk and the control disk are combined into a single disk.

The present invention also provides a disk in which the PCR disk and the disk for electrophoresis are combined.

Claims (46)

A prep chamber for performing a preparation process for preparing a DNA sample from a sample, one or more PCR chambers for performing a polymerase chain reaction (PCR) process on the DNA sample, a trech chamber for collecting debris from the prep chamber or a PCR chamber A rotatable solid substrate comprising a buffer chamber for storing a buffer solution which may be provided to the preparation chamber or the PCR chamber, a flow path formed between the chambers, and a flow path formed between the chambers and a hole formed in the flow path; And PCR disk device including a valve for opening and closing the hole. A prep chamber for performing a preparation process for preparing a DNA sample from a sample, one or more PCR chambers for performing a polymerase chain reaction (PCR) process on the DNA sample, a trech chamber for collecting debris from the prep chamber or a PCR chamber A rotatable solid substrate comprising a buffer chamber for storing a buffer solution which may be provided to the preparation chamber or the PCR chamber, a flow path formed between the chambers, and a flow path formed between the chambers and a hole formed in the flow path; And PCR disk including a valve for opening and closing the hole; A control disc including a control unit for supplying a power or control signal necessary for the PCR disc; And PCR disk apparatus including an interface unit for connecting the PCR disk and the control disk. The method according to claim 1 or 2, Wherein said solid substrate is selected from a material selected from plastic and silicon wafers. The method according to claim 1 or 2, The valve is cylindrical magnet particles; And an electromagnet which induces a repulsive force or a suction force on the cylindrical magnet particles. The method of claim 2, The interface unit includes a plurality of control signal connection units for supplying a control signal to the PCR disk; And a power connection part for supplying power to the PCR disc. The method of claim 2, The control disk further comprises a power supply connecting portion for receiving power from the external power supply by a brush or a non-contact interface. The method of claim 2, The control disk is a PCR disk device, characterized in that configured by a circuit integrated on a PCB (Printed Circuit Board) or a silicon wafer. The method according to claim 1 or 2, Fluid flow is a PCR disk device characterized in that the centrifugal force induced in the PCR disk device and the valve opening and closing. The method according to claim 1 or 2, The PCR disk device further includes a contactless interface consisting of one of an infrared interface, an optical interface, and a wireless interface, PCR disc device, characterized in that for transmitting the process progress and analysis results of the PCR disc device to the external central control device or storage device through the contactless interface. The method according to claim 1 or 2, The PCR disc can be remotely connected to a specialist doctor and hospital according to the protocol, analysis algorithms, standard control values for reading, bioinformatics information, device driver software, patient education information for clinical analysis, diagnosis results and diagnosis results. PCR disk device, characterized in that it further comprises a memory for storing a web site or various links, or personal encrypted information. The method of claim 2, The PCR disc device, characterized in that the control disc and the PCR disc are combined into one integrated PCR disc. The method of claim 11, The integrated PCR disk is made of a silicon wafer material, characterized in that the control unit, the memory, the non-contact interface, the electrode plate or electromagnet for controlling the valve, the electronic circuit including the circuit pattern, the chamber and the channel are integrated on the silicon wafer. PCR disk unit. A prep chamber for performing a preparation process for preparing a DNA sample from a sample, one or more PCR chambers for performing a polymerase chain reaction (PCR) process on the DNA sample, and a buffer solution that can be provided to the PCR chamber. A rotatable PCR disc including a buffer chamber, a flow path formed between the chambers through which fluid can flow, a hole formed in the flow path, and a valve for opening and closing the hole; A tray allowing loading of the PCR disc; A control unit for supplying a control signal necessary for the PCR disk; A motor for rotating the PCR disc; A circuit board having a central control device and a storage device; And A body supporting the PCR disc driver, And the circuit board is connected to the PCR disc driver body, and the central control unit controls the motor to rotate or stop the PCR disc. The method of claim 13, The PCR disc driver apparatus further comprises a non-contact interface means for sending the PCR process progress and analysis results to the central control unit or storage device or to receive a command from the central control unit. The method of claim 13, A PCR disc driver device further comprising an optical pickup device for reading a normal optical disc. The method of claim 14, And a PCR disc detecting means for determining whether the disc loaded in the tray of the PCR disc driver device is a normal optical disc or a PCR disc. The method of claim 16, And the PCR disc detection means notifies the central controller that the currently loaded disc is a PCR disc by wirelessly transmitting a PCR disc detection signal to the central control unit through the contactless interface. The method of claim 17, The central controller determines whether a normal optical disk or a PCR disk, In the case of a conventional optical disk, the contents read from the disk are transferred from the optical pickup device to the storage device, or the contents are written to the optical pickup device, and various control signals necessary for read / write are provided to the optical pickup device. Performing a normal operation for the optical disc, including the process of In the case of a PCR disk, the PCR disk driver device characterized in that for transmitting a control command signal for controlling the PCR disk through the contactless interface. The method of claim 13, The PCR disk driver device includes a protocol, analysis algorithms, standard control values for reading, bioinformatics information, device driver software, patient education information for clinical analysis, diagnosis results, and a remote doctor and a hospital. PCR disk driver device characterized in that it further comprises a memory or storage means for storing a web site and various links, or personal encryption information that can be connected to. The method according to claim 1 or 2, The prep chamber includes a lysis buffer solution for breaking up cells and extracting DNA, and a ferromagnetic bead having an affinity with the extracted DNA. The method according to claim 1 or 2, And a plurality of PCR chambers, wherein each of the plurality of PCR chambers includes primers of the same or different types or comprise a plurality of primers. A rotatable solid substrate comprising a flow path through which a fluid can flow, a chamber for storing a buffer solution or performing various chemical processes, and a hole connecting the flow path; And It includes a valve for opening and closing the hole, The chamber includes one or more preparation chambers for performing a preparation process for preparing a DNA sample from the sample; One or more PCR chambers for carrying out a PCR process or storing enzymes and buffer solutions including polymerases and primers required for the PCR process; And a plurality of out chambers for collecting the PCR products obtained by the PCR process, wherein the chambers are connected to the solid substrate to perform a series of PCR processes. The method of claim 22, And the output chamber includes an outlet for enabling the collection of PCR products by pipettes or syringes. The method of claim 22, The solid substrate may comprise a ladder chamber for storing a ladder marker; A control chamber for storing positive control and negative control samples; And a stain chamber. The method of claim 22, The PCR disk is a PCR disk device, characterized in that integrated with the disk for electrophoresis (Electrophoresis). The method of claim 25, And the output chambers are connected in a one-to-one correspondence with an input chamber of an electrophoresis disk. The method of claim 26, The electrophoresis disk is a PCR disk device comprising an electrode for applying a voltage to a gel plate (gel plate) consisting of agarosde gel (agarosde gel) or polyacrylamide gel (polyacrylamide gel) . The method of claim 27, The PCR disk device further comprises an optical measuring device including a UV (Ultra Violet) device and a camera to measure, store, display and transmit the band (band) formed on the gel plate. A sample analysis method using the PCR disk device of claim 28, A preparation step for preparing a DNA sample from the sample; A PCR step for amplifying DNA; Collecting the PCR product obtained by the PCR step in an out chamber; Transferring the ladder marker and positive control or negative control to a corresponding output chamber; Moving contents of the output chamber to an input chamber of the electrophoretic disk; Incubating with applying voltage to the electrode for electrophoresis; And Measuring, storing, displaying, or transmitting a band formed on the gel plate by the UV device and the camera. A blood analysis method using the PCR disk device of claim 20, Injecting blood through a sample inlet installed in the prep chamber; An incubation step for depositing DNA extracted from blood onto the ferromagnetic beads; Allowing the ferromagnetic beads to settle and then opening the valve and slowly rotating the PCR disc apparatus to flush out the debris from the cell destruction into the trash chamber; Injecting a washing buffer into the preparation chamber, allowing the ferromagnetic beads to settle, and then slowly rotating the PCR disc device while slowly opening the valve to wash away the debris generated during cell destruction into the trech chamber; And Introducing distilled water or resuspension buffer into the preparation chamber and heating or heating the preparation chamber by a heater means to separate or resuspend DNA attached to the ferromagnetic beads. A DNA analysis method using the PCR disk device of claim 20, Moving the DNA obtained by the preparation process into a PCR chamber; And And amplifying the DNA by repeating the PCR cycle by using a heater means for heating the PCR chamber and cooling by the rotation of the temperature sensor and the disk. The method of claim 31, wherein If the sample is not injected into the preparation chamber to start the PCR process, the method further comprises the step of ejecting (eject) or notifying the user of the warning message. The method according to claim 1 or 2, PCR disk device further comprises an impedance measuring device for checking whether the sample is injected into the preparation chamber. The method of claim 33, The impedance measuring device is a PCR disk device, characterized in that consisting of an interdigitated array. The method of claim 13, If the user receives an eject or stop signal from the PCR disk during the PCR process, the user can ignore the user's signal and proceed with the PCR process. And a password request unit for receiving a user's eject or stop signal and performing a corresponding process if the password is correct. The method of claim 13, PCR disc driver device further comprises a usability determination unit that can determine whether the availability of the PCR disk each time the loading of the PCR disk by the information and the expiration date information on how many times the disk used the PCR disk; . The method of claim 13, And a light emitting diode for indicating that the loaded disk is a PCR disk. The method of claim 13, And a liquid crystal display for displaying the progress of the PCR disk driver apparatus according to the preparation process, the PCR process, and the electrophoresis process in a percent or bar graph or a pie graph. The method of claim 30, And said heater means is a heater composed of an electrode pattern or an air core coil winding embedded in a preparation chamber. The method of claim 31, wherein And a remote diagnosis step of transmitting the PCR analysis result using the existing internet network. The method of claim 31, wherein And notifying the user that the PCR disk device that has passed the expiration date is unusable. The method of claim 31, wherein The heater means and the temperature sensor is an analysis method, characterized in that embedded in the PCR chamber. The method according to claim 1 or 2, And a memory storing information on how many times the PCR disk has been used, expiration date information, and disease information to be diagnosed. delete delete delete

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