KR102596867B1 - Analysis Device for Sample Analysis Chip and Sample Analysis Method using the same - Google Patents

Abstract

The present invention relates to an analysis device for a sample analysis chip, and more specifically, to an analysis device in which the preprocessing, gene amplification, and analysis processes in the sample analysis chip are fully automated.

Description

Analysis device for sample analysis chip and sample analysis method using the analysis device {Analysis Device for Sample Analysis Chip and Sample Analysis Method using the same}

The present invention relates to an analysis device on a chip for sample analysis and a method of analyzing a sample using the analysis device.

Genetic diagnosis is essential for diagnosing bacterial or viral diseases.

Genetic diagnosis consists of a total of four steps: sample collection step, sample preprocessing step (DNA or RNA extraction), gene amplification step, and detection step.

However, in the past, each step is performed by separate equipment or devices, so expensive analysis equipment and a large amount of samples are required, analysis takes a lot of time, there is a high possibility that the sample will be contaminated during diagnosis, and there is a high possibility that the sample will be contaminated during diagnosis. There is a problem that rapid diagnosis is difficult.

To solve these problems, an integrated genetic analysis device using a microchip based on microfluidics has recently been developed. It is called a lab on a chip, meaning that the laboratory environment is implemented on a single chip.

However, conventional integrated genetic analysis devices also have complex chip structures, high manufacturing costs due to metal electrode patterning and silicon/glass substrate-based structures, operational complexity due to the need for external inlet pumps and multiple tube systems, and highly integrated chip operation. There are problems such as low reproducibility of the device, lack of automation in system operation, and difficulty in on-site diagnosis due to limitations in miniaturization, so improvements are required.

As a prior art, Korean Patent Publication No. 10-2020-0064466, invented and published by the present inventor, has been used. It relates to a chip for sample analysis, and uses the principle that when a sample is injected, it passes through a zig-zag shaped capture passage and the analyte contained in the sample is captured by beads provided in the capture passage. However, there was a problem in that the efficiency of extracting the analyte from the sample was greatly reduced due to the non-uniform space between the beads. In addition, a configuration is proposed in which an oil loading unit is provided and the liquid oil stored in the oil loading unit is injected into the front of the reaction chamber, thereby preventing evaporation of the mixture introduced into the reaction chamber. However, the oil stored in the oil loading unit is liquid. Therefore, even if the outlet of the oil loading part is sealed with a seal such as paraffin wax, there is a problem of oil leaking when the sample analysis chip is rotated, causing a problem of contamination by oil even before the sample is introduced into the reaction chamber. did.

Korean Patent Document No. 10-1965963 (2019.08.13) Korean Patent Document No. 10-1986464 (2019.06.05) Korean Patent Document No. 10-2076809 (2020.02.17) Korean Patent Publication No. 10-2020-0064466 (2020.06.08) Korean Patent Publication No. 10-2018-0128054 (2018.11.30) Korean Patent Document No. 10-1091906 (2011.12.02) Korean Patent Publication No. 10-2009-0112560 (2009.10.28)

The purpose of the present invention is to provide an integrated micro device that can implement all genetic diagnostics on one chip.

Additionally, the purpose of the present invention is to provide a chip with improved capture efficiency of analyte substances contained in a sample compared to a conventional sample analysis chip.

In addition, in the present invention, solid wax is stored in the wax storage unit and does not leak out until the wax storage unit is heated, so the wax flows out before the pretreated sample is introduced into the reaction chamber, thereby reducing the accuracy of analysis. The purpose is to provide a chip that solves the problems of conventional samples analysis chips.

In addition, the purpose of the present invention is to provide an analysis device in which preprocessing of a sample analysis chip, gene amplification, and analysis processes are fully automated within a single analysis device.

In one embodiment of the present invention to solve the above problems, the sample storage unit 110 is located radially outside the sample storage unit 110, and the sample introduced into the sample storage unit 110 undergoes pretreatment. A sample analysis chip 100 is installed, which includes a preprocessing unit and a reaction chamber 180 located radially outside the preprocessing unit and into which the sample pretreated by the preprocessing unit is introduced, forming the reaction chamber 180. A device for amplifying substances in the sample and analyzing the sample, comprising: a chip installation unit 320 on which the sample analysis chip 100 is seated and a motor 322 that rotates the sample analysis chip 100; A heating device configured to be able to move closer to or away from the reaction chamber 180 while being aligned vertically with respect to the reaction chamber 180, and including a first heater 331 that heats the reaction chamber 180. An apparatus is provided, including a sensor unit 350 that is aligned vertically with respect to a unit 330 and the reaction chamber 180 and is capable of detecting fluorescence generated in the reaction chamber 180.

In one embodiment, the sample analysis chip 100 communicates with the reaction chamber 180 so as to be located radially inside the reaction chamber 180, and has solid wax inside. It further includes a stored wax storage unit 190, wherein the heating unit 330 is installed to be spaced apart from the first heater 331 and is vertically aligned with the wax storage unit 190 to form the wax storage unit 190. It may further include a second heater 332 that heats.

In one embodiment, the heating unit 330 may further include a Peltier element 333 connected to the first heater 331.

In one embodiment, the heating unit 330 may be installed on the upper and lower sides of the reaction chamber 180, respectively.

In one embodiment, a plurality of reaction chambers 180 are provided, wherein the plurality of reaction chambers 180 are formed along the circumferential direction of the sample analysis chip 100, and the first heater 331 is provided with the plurality of reaction chambers 180. It may be formed to extend along the circumferential direction.

The area of the first heater 331 may be configured to cover at least all of the plurality of reaction chambers 180, and the area of the second heater 332 may be configured to cover at least the wax chamber 190. .

In one embodiment, the thickness of the portion of the sample analysis chip 100 where the reaction chamber 180 is formed may be thinner than other portions.

In one embodiment, based on the fluorescence intensity detected by the sensor unit 350, an operation processing unit ( 360) may be further included.

In addition, the present invention is an analysis method using the above device, including (a) introducing a sample into the sample storage unit 110, (b) rotating the sample analysis chip in a preset direction at a preset rotation speed. The sample introduced in step (a) is pretreated, and the pretreated sample is introduced into the reaction chamber 180, (c) the direction in which the heating unit 330 approaches the sample analysis chip 100. moving to, (d) the second heater 332 is heated to a preset temperature and the wax stored in the wax storage unit 190 is melted, (e) the sample analysis chip 100 is Rotating in a preset direction at a preset rotation speed, the wax melted in step (d) is injected and dispensed into the input channel 174 located radially inside the reaction chamber 180, (f) An analysis method comprising heating the first heater 331 to follow a preset reaction temperature cycle and (g) detecting fluorescence generated in the reaction chamber 180 by the sensor unit 350. provides.

In one embodiment, step (a) includes inserting the sample into the sample storage unit 110 by coupling the cartridge 200 storing the sample to the sample analysis chip 100, After step (b) and before step (c), the sample analysis chip 100 is rotated so that the second heater 332 is vertically aligned with the wax storage unit 190, and the ( In step f), the first heater 331 performs polymerase chain reaction (Polymerase Chain Reaction, PCR), Reverse Transcription Polymerase Chain Reaction (RT-PCT), and Isothermal Amplification Reaction. A step of amplifying the sample introduced into the reaction chamber 180 by heating to follow the temperature cycle of one or more reactions may be further included.

In one embodiment, step (f) controls the intensity and direction of the current flowing through the Peltier element 333, so that the first heater 331 connected to the Peltier element 333 is set to a preset reaction temperature. It may further include heating to follow the cycle.

In one embodiment, after step (f) and before step (g), the sample analysis chip 100 is rotated so that the sensor unit 350 and the reaction chamber 180 are aligned vertically. More may be included.

In one embodiment, after step (g), (h) the operation processing unit 360 determines the presence or absence of the analyte in each reaction chamber 180 based on the fluorescence intensity detected in step (g). A step of determining whether the sample is infected with a disease may be further included.

In one embodiment, in step (h), the calculation processing unit 360 determines the types of primers provided in the plurality of reaction chambers 180 and each reaction chamber 180 detected by the sensor unit 350. ) may further include determining whether a substance to be analyzed is present in the reaction chamber 180 or whether the sample is infected with a disease, using the number of combinations of fluorescence intensities.

In one embodiment, a material having a reference fluorescence intensity is stored in advance in at least one reaction chamber among the plurality of reaction chambers 180, and in step (h), the operation processing unit 360 is configured to use the sensor. Among the fluorescence intensities of each reaction chamber 180 detected in the unit 350, the reaction chamber 180 matching the reference fluorescence intensity is determined as the reference reaction chamber, and other reaction chambers 180 are selected based on the determined reference reaction chamber. The step of determining the location may further be included.

According to the present invention, it is possible to implement all genetic diagnosis on one chip.

Additionally, compared to a conventional sample analysis chip, the capture efficiency of the analyte contained in the sample is improved.

In addition, solid wax is stored in the wax storage unit and does not leak out until the wax storage unit is heated, so the wax leaks out before the pretreated sample is introduced into the reaction chamber, thereby reducing the accuracy of analysis in conventional sample analysis. It is possible to solve the problem of the dragon chip.

In addition, the preprocessing, gene amplification, and analysis processes of the sample analysis chip can be fully automated within one analysis device.

In addition, according to the present invention, it is possible to immediately diagnose on-site infection with pathogens that have a large social impact, such as influenza virus, avian influenza virus, MERS outbreak, Zika virus, and foot-and-mouth disease, which occur every year.

Through this, it is possible to prevent virus infection in advance and reduce human casualties and economic losses.

The advantage of genetic diagnosis, which can currently only be performed at large medical institutions such as university hospitals, can be quickly diagnosed even at small medical institutions and public health centers.

In addition, it is possible to diagnose on-site viruses/bacteria infected with food, such as food poisoning bacteria, and continuously monitor problems that occur during meal distribution, which are widespread in kindergartens and elementary/middle/high schools.

1 is a diagram for explaining a chip for sample analysis according to the present invention.
FIG. 2 is a diagram for explaining a unit process unit of the sample analysis chip of FIG. 1.
Figure 3 is a cross-sectional view for explaining the capture passage and the configuration connected to the capture passage.
FIG. 4 is a diagram for explaining a cartridge that is coupled to the sample analysis chip of FIG. 1 and injects a solution.
Figure 5 is a diagram of an analysis device in which a sample analysis chip according to the present invention is installed and can perform pretreatment, gene amplification, and analysis processes on the sample input to the sample analysis chip.
FIG. 6 is a diagram for explaining the internal configuration of the analysis device of FIG. 5.
Figure 7 is a diagram for explaining a heating unit that heats the reaction chamber and wax storage unit of the sample analysis chip.
FIG. 8 is a diagram for explaining the configuration of the second heater of the heating unit of FIG. 7.
Figure 9 is a diagram showing the heating unit, the reaction chamber of the sample analysis chip, and the wax storage unit when aligned vertically.
Figure 10 is a schematic diagram to explain how the reaction chamber and wax storage part of the sample analysis chip and the heating part of the analysis device are aligned vertically and heating is performed by a heater.
Figure 11 is a schematic diagram to explain how the reaction chamber of the sample analysis chip and the sensor unit of the analysis device are aligned vertically and detect optical signals from the sensor unit.
Figures 12A to 12L are diagrams for explaining the pretreatment and gene amplification process of samples input to the sample analysis chip using the sample analysis chip according to the present invention.
Figure 13 is a result diagram according to verification experiment 1.
Figures 14 and 15 are result diagrams according to verification experiment 2.
Figures 16 to 20 show results according to verification experiment 3.
Figures 21 and 22 are results according to verification experiment 4.
Figure 23 is a diagram of a conventional sample analysis chip.

Hereinafter, the term “gene amplification” means that the gene to be analyzed is amplified. Examples of gene amplification include PCR (Polymerase Chain Reaction), Real-time PCR, and Isothermal Amplification Reaction, and are not limited to and include any reaction that amplifies the gene to be analyzed. Explain in advance that this will happen.

Chip for sample analysis

With reference to the attached drawings, the sample analysis chip 100 according to the present invention will be described in detail.

Referring to FIG. 1, the sample analysis chip 100 may have a disk shape, and a first hole H is formed in the center. The first through hole (H) is coupled to the chip analysis device 300 for sample analysis, which will be described later, and corresponds to the rotation axis when rotating. A second through hole (h) is formed on the radial outer side of the first through hole (H), and a vibration prevention unit 400, which will be described later, is coupled to the second through hole (h) to rotate the sample analysis chip 100 at high speed. This solves the problem of the sample analysis chip 100 vibrating or the solution leaking out of the cartridge 200. A detailed explanation of this will be provided later.

The sample analysis chip 100 is largely composed of three layers. That is, the upper and lower surfaces of disc-shaped PMMA with a preset thickness, for example, 3 mm, are processed using a CNC milling machine to form a groove-shaped pattern for each configuration, and then a PSA film (PSA) is applied to the upper and lower surfaces. It can be manufactured by adhering a film).

That is, the sample analysis chip 100 includes a disk 10, a first film layer 20 adhered to the upper surface of the disk 10, and a third film layer 30 adhered to the lower surface of the disk 10. In addition, the components described later are patterned in the form of grooves on the upper or lower surface of the disk 10. The first film layer 20 covers the top of each patterned component, and the second film layer 30 covers the bottom. I do it.

The sample analysis chip 100 according to the present invention includes a plurality of unit processing units 100a sequentially arranged along the circumferential direction. FIG. 1 shows an embodiment in which two unit process units are formed on a chip for sample analysis, and FIG. 2 is a diagram for explaining one unit process unit in detail.

The unit process unit 100a is formed in a pattern on the disk 10 and includes a sample storage unit 110, a capture passage 120, a cleaning liquid storage unit 130, a delay unit 140, a cocktail storage unit 150, It includes an eluent storage unit 160, a connection chamber 170, a waste liquid chamber 171, a collection chamber 172, a reaction chamber 180, and a wax chamber 190.

Injection ports 111, 131, 151, and 161 are formed in the sample storage unit 110, the cleaning liquid storage unit 130, the cocktail storage unit 150, and the eluent storage unit 160, respectively, and the sample according to the present invention is formed. The cartridge 200 is mounted on the analysis chip 100 so that each solution can be injected.

That is, a sample inlet 211, a cleaning solution inlet 231, a cocktail inlet 251, and an eluent inlet 261 are formed in the cartridge 200, and a Spaces 210, 230, 250, and 260 are formed inside, and each solution is stored in this space. When the sample analysis chip 100 is rotated, the solutions stored in the cartridge 200 are stored in the sample storage unit 110, It may be put into the washing liquid storage unit 130, the cocktail storage unit 150, and the eluent storage unit 160.

The sample storage unit 110 is located radially inside the capture passage 120 and loads the sample introduced from the cartridge 200 into the capture passage 120.

The loaded sample flows into the waste liquid chamber 171 through the capture passage 120 and the connection chamber 170 by centrifugal force when the sample analysis chip 100 rotates. More specifically, the sample introduced from the cartridge 200 into the sample inlet 111 sequentially passes through the inlet 112 and the sample inlet channel 113 and is injected into the capture passage 120. At the connection point between the input unit 112 and the sample input channel 113, and at the connection point between the sample input channel 113 and the capture passage 120, there are portions 114, 121 that are not covered by the first film layer 20. ) exists, in other words, external air flows in through the parts 114 and 121, allowing the sample introduced into the sample storage unit 110 to move more quickly to the capture passage 120. Therefore, before the sample reaches the capture passage 120, the washing liquid introduced into the washing liquid storage unit 130 reaches the capturing passage 120, or before all samples pass through the capturing passage 120, the washing liquid storage unit ( The problem of the washing liquid introduced into 130) reaching the capturing passage 120 can be prevented.

The capture passage 120 captures the analyte target from the sample loaded through the sample storage unit 110. An inlet 122 and an outlet 123 are provided at both ends of the capture passage 120, respectively. The inlet 122 of the capture passage 120 communicates with the sample storage unit 110, the washing liquid storage unit 130, and the eluent storage unit 160, which are located radially inside the capture passage 120. The outlet 123 of 120 communicates with the connection chamber 170 located radially outward from the capture passage 120. Therefore, as the sample analysis chip 100 rotates, the sample, cleaning solution, and eluent may flow into the connection chamber 170.

Meanwhile, the components of the sample storage unit 110, the cleaning liquid storage unit 130, and the eluent storage unit 160, which communicate with the inlet 122 of the capture passage 120, are provided in the form of a groove on the upper surface of the disk 10. and a connection chamber 170 in communication with the outlet 123 of the capture passage 120, a waste liquid chamber 171, a collection chamber 172, an input channel 174, a reaction chamber 180, and a wax storage portion ( The components 190) are provided in the form of grooves on the lower surface of the disk 10. In addition, the capture passage 120 is provided in the vertical direction of the disk 10 for communication between the components provided in the form of grooves on the upper surface of the disk 10 and the components provided in the form of grooves on the lower surface of the disk 10. It is provided in the form of a groove extending to (see Figure 3).

A membrane (f) for capturing the analyte target is provided on the capture passage 120. The membrane (f) according to the present invention may be a glass filter, and the glass filter may be provided with a plurality of silica beads in the form of a film. Specifically, one or more glass filters may be installed on the capture passage 120. More preferably, a glass filter made of a triple layer of a plurality of silica beads is installed on the capture passage 120, so that the analyte can be captured from the sample with high efficiency (see FIGS. 14 and 15). The silica beads of the glass filter have a negative charge, and if the glass filter is configured to include a positively charged salt, such as a guanidine salt, the silica bead - guanidine salt - the negatively charged analyte (DNA, etc.) Through the combination, analyte substances such as DNA can be captured in a glass filter.

The washing liquid storage unit 130 is in the form of a chamber and stores washing liquid for washing (removing) substances other than the analyte target captured in the capture passage 120. The cleaning liquid storage unit 130 is located radially inside the capturing passage 120 and is connected to the inlet 122 of the capturing passage 120. Therefore, as the sample analysis chip 100 rotates, the cleaning solution stored in the cleaning solution storage unit 130 flows into the capturing passage 120 to wash (remove) the remaining substances except for the analyte captured in the capturing passage 120. )can do.

A delay unit 140 may be provided between the cleaning liquid storage unit 130 and the capture passage 120. The delay unit 140 is configured to delay passage of the cleaning solution from the cleaning solution storage unit 130 so that it reaches the capture passage 120 later than the sample.

The delay unit 140 includes an inlet channel 141, a first delay chamber 142, and a delay channel 143.

The inflow channel 141 connects the cleaning liquid storage unit 130 and the first delay chamber 142, and allows the cleaning liquid introduced into the cleaning liquid storage unit 130 to flow into the first delay chamber 142. The inlet channel 141 includes a first passage 141a extending away from the first delay chamber 142 in a first circumferential direction from the outlet 132 of the cleaning liquid storage unit 130 and a first passage 141a extending from the outlet 132 of the cleaning liquid storage unit 130. It includes a second passage 141b extending close to the first delay chamber 142 in a second circumferential direction opposite to the first circumferential direction.

The first delay chamber 142 stores the cleaning solution heading from the cleaning solution storage unit 130 to the capture passage 120.

The delay channel 143 is provided between the first delay chamber 142 and the capture passage 120 to delay the passage of the cleaning liquid. To this end, the delay channel 143 includes a first delay passage 143a extending radially inward and a second delay passage 143b connected to the first delay passage 143a and extending radially outward. . Therefore, since the cleaning solution is not introduced into the capture passage 120 through the delay channel 143 until the cleaning solution stored in the first delay chamber 142 reaches a preset amount, the cleaning solution is released into the capture passage 120 more than the sample. ) is reached with a delay. Therefore, it is possible to prevent the problem of the washing liquid flowing into the capturing passage 120 before the material to be analyzed is captured.

A cocktail is introduced into the cocktail storage unit 150. Here, the cocktail is a gene amplification enzyme (e.g., DNA polymerase) or MgCl ₂ salt. May contain materials required for PCR or RT-PCR.

The cocktail storage unit 150 is connected to the connection chamber 170 by a cocktail introduction channel 153.

The cocktail introduction channel 153 includes a third passage 153a extending radially inward and a fourth passage 153b connected to the third passage 153a and extending radially outward. Therefore, the cocktail may not flow into the connection chamber 170 when the sample analysis chip 100 rotates, but may flow into the connection chamber 170 when the sample analysis chip 100 stops. Therefore, after the sample and the cleaning solution are all introduced into the capture passage 120, the cocktail may be sequentially introduced into the connection chamber 170. The cocktail introduction channel 153 described above may have a hydrophilic coating structure.

The eluent storage unit 160 is injected with an eluent for separating the analyte captured in the capture passage 120. The eluent storage unit 160 is located radially inside the capture passage 120 and communicates with the capture passage 120. Therefore, as the sample analysis chip 100 rotates, the eluent stored in the eluent storage unit 160 flows into the capture passage 120 to separate the analyte captured in the capture passage 120 from the membrane f. can do.

The eluent storage unit 160 is connected to the capture passage 120 and the eluent introduction channel 163.

The eluent introduction channel 163 includes a fifth passage 163a extending radially inward and a sixth passage 163b connected to the fifth passage 163a and extending radially outward. Therefore, when the sample analysis chip 100 rotates, the eluent does not flow into the capture passage 120, but when the sample analysis chip 100 stops, the eluent may flow into the capture passage 120. Therefore, after the sample and the cleaning solution have all flowed into the capture passage 120, the eluent may sequentially flow into the capture passage 120. The eluent introduction channel 163 described above may have a hydrophilic coated structure.

The connection chamber 170 is located radially outside the capturing passage 120 and the cocktail storage unit 150, and is connected to the capturing passage 120 and the cocktail storage unit 150.

A waste liquid chamber 171 and a collection chamber 172 are located radially outside the connection chamber 170.

The sample and washing liquid that passed through the capture passage 120 flow into the waste liquid chamber 171, and the eluent that passed through the capture passage 120 and the cocktail introduced into the connection chamber 170 flow into the collection chamber 172.

The connection chamber 170 is connected to the waste liquid chamber 171 by a connection channel. The connection channel may be provided with one or more capillary valves 170a, and the capillary valve 170a prevents the solution stored in the waste liquid chamber 171 from being introduced back into the connection chamber 170 through the connection channel. The connection chamber 170 and the capillary valve 170a described above may have a hydrophobic coating structure.

The waste liquid chamber 171 stores the sample and cleaning liquid that passed through the capture passage 120. Unnecessary substances other than the substance to be analyzed can be stored, and a super absorbent polymer that absorbs the sample that has passed through the capture passage 120 can be attached thereto.

The collection chamber 172 stores and mixes an eluent and a cocktail containing the analyte.

A distribution portion is formed radially outside the collection chamber 172. A mixed solution of the eluent and the master mix may be introduced into the distribution unit.

A mixture introduction channel 173 is formed between the collection chamber 172 and the distribution unit, and the mixture introduction channel 173 is connected to a seventh passage 173a extending radially inward and the seventh passage 173a. and includes an eighth passage 173b extending outward in the radial direction. Therefore, when the sample analysis chip 100 rotates, the mixture may not flow into the distribution unit, but may flow into the distribution unit when the sample analysis chip 100 stops. The above-described mixture introduction channel 173 may have a hydrophilic coated structure.

The distribution unit includes an input channel 174, a connection channel 175, and a reaction chamber 180.

The input channel 174 is connected to the collection chamber 172 by the mixture introduction channel 173, and distributes the mixture passing through the collection chamber 172 to at least one reaction chamber 180. To this end, the input channel 174 extends a predetermined length along the circumferential direction and has an aliquoting structure.

The input outlet formed in the input channel 174 is connected to the reaction chamber 180 and the connection channel 175, respectively. The connection channel 175 extends from each input outlet formed in the input channel 174 to each reaction chamber 180, and is narrower than the width of the input outlet formed in the input channel 174. Additionally, the length of each connection channel 175 may be made shorter as it is positioned further away from the collection chamber 172 in the circumferential direction.

The reaction chamber 180 is located radially outside the connection channel 175 and corresponds to the input outlet formed in the input channel 174. The reaction chamber 180 receives the distributed mixture through the input channel 174, and PCR or RT-PCR is performed on the distributed mixture depending on the analyte. Different primers are stored in the reaction chamber 180 to detect the analyte contained in the distributed mixture. In addition, a material having a reference fluorescence intensity may be stored in one or more reaction chambers among the plurality of reaction chambers 180, and the information on the reference fluorescence intensity is pre-stored in the analysis device 300 to be described later, and is later stored in the calculation processing unit ( 360), by determining the reaction chamber showing the reference fluorescence intensity as the reference reaction chamber, the location of the other reaction chamber can be specified.

The wax chamber 190 is located radially inside the distribution unit and is connected to the input channel 175 through the wax introduction channel 191. Solid wax is stored in the wax chamber 190, and after the mixture is distributed to the reaction chamber 180, it is added to prevent evaporation of the mixture. After the mixture is introduced into the reaction chamber 180, when heat exceeding a preset temperature is applied from the outside, the melted wax (oil) is introduced into the input channel 174 and the connection channel 175 to the reaction chamber 180. Evaporation of the added mixture can be prevented.

Meanwhile, since solid wax is stored in the wax chamber 190, unless the wax chamber 190 is heated, it is not input into the input channel 175 even if the sample analysis chip 100 rotates or stops after rotation. Therefore, by storing oil in a liquid state in the conventional sample analysis chip, the problem of oil flowing out and mixing with the mixture when the sample analysis chip is rotated is prevented.

As described above, the vibration prevention unit 400 is configured to be coupled to the second hole h of the sample analysis chip 100. For this purpose, a coupling protrusion 401 that engages the second through hole (h) is formed to protrude from the lower part of the vibration prevention unit 400.

The vibration prevention unit 400 is coupled to the sample analysis chip 100 after the cartridge 200 is coupled to the sample analysis chip 100. The vibration prevention unit 400 has an area large enough to cover the entire cartridge 200 coupled to the sample analysis chip 100, and through this, when the sample analysis chip 100 rotates, the cartridge 200 The problem of internal solution leaking to the outside due to vibration is prevented.

Sample analysis method

With reference to FIGS. 12A to 12L, the sample analysis method according to the present invention will be described in detail.

First, for sample analysis, each injection port of the cartridge 200 is connected to the sample storage unit 110, the cleaning liquid storage unit 130, the cocktail storage unit 150, and the eluent storage unit of the sample analysis chip 100 ( 160) (FIG. 12b). Accordingly, the solution stored in each solution storage unit of the cartridge 200 is injected into the storage unit of the sample analysis chip 100, and the sample is injected into the sample storage unit 110.

Next, the sample analysis chip 100 rotates in the first direction at a first rotation speed W ₁ (eg, 5000 rpm) for a preset time (eg, 90 seconds). As it rotates, the sample injected into the sample storage unit 110 passes through the capture passage 120, the analyte contained in the sample is captured by the membrane (f), and the remaining materials not captured are connected to the connection chamber ( 170) and is introduced into the waste liquid chamber 171.

As the sample analysis chip 100 continues to rotate, the washing liquid flowing into the first delay chamber 142 and stored therein passes through the capture passage 220 and is captured by the membrane f, but is not an analyte material. After cleaning, the washed material and cleaning liquid pass through the connection chamber 170 and are introduced into the waste liquid chamber 171 (FIG. 12c).

Next, the sample analysis chip 100 is stopped for a preset time (e.g., 10 seconds), and when the sample analysis chip 100 is stopped, the centrifugal force due to rotation disappears and the cocktail storage unit is moved by capillary force. The cocktail stored in 150 and the eluent stored in the eluent storage unit 160 pass through the cocktail introduction channel 153 and the eluent introduction channel 163, respectively (FIG. 12d).

Next, the sample analysis chip 100 is rotated around the first hole H in a second direction opposite to the first direction at a first rotation speed W ₁ for a preset time (for example, 30 seconds). It rotates. As the rotation progresses, the cocktail that has passed through the cocktail introduction channel 153 is injected into the connection chamber 170, and the eluent that has passed through the eluent introduction channel 163 passes through the capture passage 120 and is captured in the membrane (f). The target material is separated from the membrane (f), and the eluent containing the analyte material passes through the connection chamber 170 and is introduced into the collection chamber 172 (FIG. 12e).

Next, the sample analysis chip 100 alternates between the first direction and the second direction at a second rotation speed W ₂ (e.g., 2000 rpm) for a preset time (e.g., 10 seconds). Rotate repeatedly. As the rotation rotates, the eluent containing the analyte injected into the collection chamber 172 and the cocktail are mixed together to create a mixture (FIG. 12f).

Next, the sample analysis chip 100 is stopped for a preset time (for example, 35 seconds), and when the sample analysis chip 100 is stopped, the centrifugal force due to rotation disappears and the collection chamber 172 is formed by capillary force. ) passes through the mixture introduction channel 173 (FIG. 12g).

Next, the sample analysis chip 100 rotates in the second direction at a third rotation speed W ₃ (eg, 1000 rpm) for a preset time (eg, 35 seconds). As the mixture rotates, the mixture moves a predetermined length along the circumferential direction and is injected into the injection outlet formed on the outside of the injection channel 174 having a dispensing structure. That is, the mixture is sequentially injected into the input outlet of the input channel 174 along the circumferential direction from the point where the collection chamber 172 and the input channel 174 are connected. Additionally, the surplus mixture that is not injected through the input outlet of the input channel 174 is injected into the surplus mixture storage chamber 176 (FIG. 12h).

Next, the sample analysis chip 100 rotates in the second direction at a first rotation speed W ₁ (eg, 5000 rpm) for a preset time (eg, 10 seconds). According to rotation, the mixture injected into each input outlet of the input channel 174 is injected into each reaction chamber 180 through the connection channel 175 (FIG. 12i).

Next, the sample analysis chip 100 is stopped for a preset time (e.g., 180 seconds), and the wax storage unit 190 is heated to melt the wax stored in the wax storage unit 190 (FIG. 12J) .

Next, the sample analysis chip 100 rotates in the second direction at a second rotation speed (eg, 2000 rpm) for a preset time (eg, 30 seconds). As the rotation rotates, the melted wax is introduced into the inlet channel 174 through the wax inlet channel 191. As the melted wax is introduced into the input channel 174, the mixture introduced into the reaction chamber 180 does not evaporate (FIG. 12k).

Next, the reaction chamber 180 is heated or cooled so that the temperature of the reaction chamber 180 follows the temperature of the preset reaction cycle. For example, the reaction chamber 180 may be heated or cooled to a temperature suitable for the PCR cycle, and through this, the analyte introduced into the reaction chamber 180 may be amplified (FIG. 12L).

Analysis device on chip for sample analysis

5 to 11, the analysis device 300 of the sample analysis chip 100 according to the present invention will be described in more detail.

Referring to Figures 5 and 6, the analysis device 300 according to the present invention includes a housing 310, a chip installation unit 320, a heating unit 330, a driving unit 340, a sensor unit 350, and an arithmetic processing unit. It includes 360 and an output unit 370.

The housing 310 forms the exterior of the analysis device 310, and the chip installation unit 320, heating unit 330, driving unit 340, sensor unit 350, and calculation unit are located in the internal space of the housing 310. A processing unit 360 and an output unit 370 are installed.

The chip installation unit 320 is configured to seat the sample analysis chip 100. Specifically, the chip installation unit 320 may be accommodated in the housing 310, and may also be exposed to the outside for installation of the sample analysis chip 100. That is, the chip installation unit 320 can be moved in a direction away from or closer to the housing 310 (movable horizontally or vertically with respect to the ground), and the chip installation unit 320 is located inside the housing 310. ) A chip installation unit driver (not shown) that moves the chip is installed. The chip installation unit driver (not shown) may be, for example, a step motor, and the chip installation unit 320 can be moved between positions stored in the housing 310 or exposed to the outside through step angle control of the step motor. possible.

The chip installation unit 320 includes a seating tray 321 on which the sample analysis chip 100 is seated.

A motor 322 is installed on the seating tray 321 to rotate the sample analysis chip 100. The upper surface of the seating tray 321 has the first hole H and the second hole H of the sample analysis chip 100. Protrusions 321a and 321b coupled to the through hole h are formed, and a motor 322 for rotating the seating tray 321 is installed at the lower portion of the seating tray 321. Therefore, as the motor 322 rotates, the sample analysis chip 100 rotates together with the seating tray 321.

The heating unit 330 is configured to heat the sample analysis chip 100. Specifically, the heating unit 330 includes a first heater 331, a second heater 332, a Peltier element 333, a heat sink 334, a fan 335, a body 336, and a first installation pillar ( 337) and a second installation pillar 338.

The first heater 331 serves to heat the reaction chamber 180 of the sample analysis chip 100. A mixture of an eluent and a cocktail containing a substance to be analyzed is stored in the reaction chamber 180. Generally, the amount of sample introduced into the sample storage unit 110 is very small, so the amount of the substance to be analyzed contained in the sample is also very small. Accordingly, in order to detect the presence or absence of a substance to be analyzed in the reaction chamber 180, it is necessary to amplify it. Different primers for amplification of the analyte target are already introduced into the reaction chamber 180, and it is necessary to perform a so-called gene amplification process using the primers and the analyte target.

The gene amplification process may be, for example, polymerase chain reaction (PCR), and generally involves a denaturation step in which DNA consisting of a double helix structure is separated into single strands by heating to a temperature of about 95°C, followed by about It can be comprised of an annealing step in which the primer and a single strand of DNA are combined by heating to a temperature of 56°C, and an extension step in which the polymerase extends the strand from the primer by heating to a temperature of about 72°C. there is. In addition, the gene amplification process may be reverse transcription-polymerase chain reaction (RT-PCR) or isothermal amplification reaction, and is not particularly limited and includes any reaction that amplifies the gene to be analyzed.

That is, the first heater 331 serves to amplify the analyte material in the reaction chamber 180 while sequentially heating to the temperature required for the polymerase chain reaction.

The first heater 331 according to the present invention may apply a method in which the first heater 331 itself is directly heated, but as shown in FIG. 7, a Peltier element ( It is also possible to apply an indirect heating method in which heat is conducted from the Peltier element 333 and the first heater 331 is heated/cooled by heating/cooling of 333). The Peltier device 333 is a device that uses the Peltier effect, and is a device in which a surface in contact with a semiconductor can be heated or cooled depending on the intensity and direction of the current flowing through the Peltier device 333. Since the Peltier device is already widely known, a detailed description will be omitted. Depending on the strength and direction of the current flowing through the Peltier element 333, the surface of the Peltier element 333 in contact with the first heater 331 may be heated or cooled, thereby heating/cooling the first heater 331. Cooling is possible. That is, the temperature of the first heater 331 can be controlled by varying the intensity and direction of the current flowing through the first wire 333a and the second wire 333b connected to the Peltier element 333.

The first heater 331 may have a curved shape so as to heat all of the plurality of reaction chambers 180 extending along the circumferential direction of the sample analysis chip 100. That is, the first heater 331 may also have a shape that extends along the circumferential direction of the sample analysis chip 100 and, as shown in FIG. 7, has an arc shape and only uses the reaction chamber 180. Can be heated/cooled selectively.

The second heater 332 serves to heat the wax chamber 190 of the sample analysis chip 100. Since the wax stored in the wax chamber 190 is in a solid state, a process of melting it into a liquid state is required. As the second heater 332 is heated, the wax in the wax chamber 190 is melted and injected into the input channel 174, and is dispensed into the input outlet of the input channel 174 as the sample analysis chip 100 rotates. do. When the wax melts, it changes into liquid oil, preventing evaporation of the mixture introduced into the reaction chamber 180.

The first heater 331 and the second heater 332 are installed to be spaced apart from each other within the heating unit 300. This is to prevent components other than those to be heated from being heated by heating each of the first heater 331 and the second heater 332. The first heater 331 is configured to heat the reaction chamber 180, and the second heater 332 is configured to heat the wax chamber 190. If one heater is heated and affects the other heaters, Since this leads to inaccurate detection results, the first heater 331 and the second heater 332 are installed spaced apart from each other. That is, the first heater 331 can be formed to cover all of the reaction chambers 180 provided in the sample analysis chip 100, and the second heater 332 is provided in the sample analysis chip 100. It is formed to cover the wax chamber 190, and the separation distance is preferably at least longer than the length of the connection channel 175. As shown in FIG. 7, the second heater 332 may have a square plate shape. Unlike the first heater 331, the second heater 332 does not require precise temperature control, so the Peltier element 333 ), and the second heater 332 itself is heated by heating of a separate heating element 332a, thereby heating the wax chamber 190.

A temperature sensor (t) may be installed in both the first heater 331 and the second heater 332, and it is possible to monitor the heating temperature of each heater in real time using the information output from the temperature sensor (t). do.

The heating unit 330 includes a heat sink 334 and a fan 335 to dissipate heat from the first heater 331, the second heater 332, and the Peltier element 333. As shown in FIG. 7, the heat sink 334 may be installed below the first heater 331, the second heater 332, and the Peltier element 333, and the fan 335 may be installed below the heat sink 334. ) is installed to be located on one side of the heat sink 334, making it possible to quickly discharge the heat of the heat sink 334 to the outside.

The body 336 is configured to have the above-described first heater 331, second heater 332, and Peltier element 333 installed, and forms the exterior of the heating unit 330. The body 336 is disposed to face the upper body 336a where the first heater 331, the second heater 332, and the Peltier element 333 are installed, and the driving unit (described later) It includes a lower body 336b connected to 340).

The upper body 336a and the lower body 336b may be connected to each other by a first installation pillar 337 and a second installation pillar 338, and on the outside of each installation pillar 337 and 338, a spring 337a, 338a) is installed. When the driving unit 340 operates, the heating unit 330 approaches the sample analysis chip 100, and the first heater 331 or the second heater 332 comes into contact with the sample analysis chip 100. In this state, when the driving unit 340 applies more force to the heating unit 330 in the direction of the sample analysis chip 100, the springs 337a and 338a contract (that is, the upper body and the lower body become closer). The first heater 331 and the second heater 332 may be completely adhered to the sample analysis chip 100. Accordingly, the heat generated from each heater can be completely applied to the sample analysis chip 100.

The driving unit 340 is connected to the heating unit 330 and moves the heating unit 330. As shown in FIG. 1, one sample analysis chip 100 includes two unit process units 100a, and as shown in FIG. 6, one analysis device 300 includes four heating units ( 330) may be included.

Two heating units 330 per unit process unit 100a may be aligned vertically with each other. That is, with the unit process unit 100a as the center, each heating unit 330 faces each other.

After the sample is introduced into the sample storage unit 110, heating by the heating unit 330 is not required until the mixture is introduced into the reaction chamber 180 according to the sample analysis method described above. When the mixture is introduced into the reaction chamber 180, a melting process of the wax stored in the wax storage unit 190 and a gene amplification process in the reaction chamber 180 are required. At this time, the driving unit 340 is connected to the heating unit 330. The heating unit 330 is moved to contact the sample analysis chip 100.

That is, the driving unit 340 moves in a downward or upward direction where the heating unit 330 approaches the sample analysis chip 100, and in a downward or upward direction where the heating unit 330 moves away from the sample analysis chip 100. The heating unit 330 is moved to do so. In the analysis device 300 according to the present invention, two driving units 340 are provided, one driving unit 340 moves the heating unit 330 located on the upper side, and the other driving unit 340 moves the heating unit 330 located on the lower side. It can be used to move the located heating unit 330.

The sensor unit 350 may be installed on the upper or lower side of the sample analysis chip 100 and is configured to detect fluorescence generated in the reaction chamber 180. As the analyte provided in the reaction chamber 180 undergoes an amplification process, its fluorescence intensity becomes stronger. When light of a specific wavelength is irradiated to each reaction chamber 180, the fluorescent substance in the reaction chamber 180 It absorbs light of a specific wavelength and enters an excited state, returning to the ground state and emitting light of a different wavelength. That is, the sensor unit 350 includes an irradiator that irradiates light of a specific wavelength to the reaction chamber 180 and a detector that detects light of a specific wavelength generated in the reaction chamber 180, and the specific wavelength detected by the detector. Based on the presence and intensity of light, it is possible to determine whether the sample contains the substance to be analyzed and whether the sample is infected with a disease.

As a measurement method of the sensor unit 350, the sensor unit 350 is aligned up and down in each reaction chamber 180, and each time it is aligned, light of a specific wavelength is irradiated and detected in the reaction chamber 180. A moving scanning method may be used in which the sensor unit 350 continuously irradiates and detects light of a specific wavelength while the sample analysis chip 100 rotates.

In addition, as an example, a material that generates strong fluorescence is manufactured to be accommodated in the reaction chamber 180 closest to the collection chamber 172 among the reaction chambers 180, and then detected through the sensor unit 350 to create a corresponding reaction chamber. It is possible to determine the location of the other reaction chamber 180 based on 180. In other words, since the reaction chamber 180 in which strong fluorescence intensity is detected is predetermined, it is possible to determine the location of the other reaction chamber 180 based on the corresponding reaction chamber 180. Through the above-described method, the calculation processing unit 360, which will be described later, can calculate in which reaction chamber 180 fluorescence was detected and at what intensity the fluorescence was detected.

The calculation processing unit 360 is configured to determine whether the sample contains a substance to be analyzed based on information detected by the detector of the sensor unit 350 and whether the sample is infected with a disease.

For example, when fluorescence is detected in any reaction chamber 180, the primers accommodated in the reaction chamber 180 (primers bind specifically according to the type of the analyte to perform a gene amplification reaction) It can be determined that the corresponding analyte substance (playing a role) is included in the sample. This is because the detection of fluorescence of a predetermined intensity means that a gene amplification reaction was performed by reacting with the primers accommodated in the reaction chamber 180.

Additionally, if fluorescence of a predetermined intensity or higher is detected in any reaction chamber 180, it may be determined that the disease is infected by the analyte corresponding to the primer contained in the corresponding reaction chamber 180. Since there is a group of diseases in which a sample cannot be considered infected simply because it contains the analyte, the corresponding analysis is performed when fluorescence of a predetermined intensity or higher (i.e., meaning that there is a lot of the analyte contained in the sample) is detected. It can be determined that there is disease infection caused by the target substance.

Sample analysis method by analysis device

First, the chip installation unit 320 protrudes out of the housing 310 by driving the chip installation unit driver (not shown), and the first through hole (H) and the second through hole (h) of the sample analysis chip 100 ) is coupled to the protrusions 321a and 321b of the seating tray 321, so that the sample analysis chip 100 is fixed to the seating tray 32.

Next, the chip installation unit 320 is stored inside the housing 310 by driving the chip installation unit driver (not shown).

Next, in accordance with the above-described “sample analysis method,” the motor 322 of the seating unit 321 rotates, and the mixture of the eluent containing the analyte target and the cocktail is introduced into the reaction chamber 180.

Next, the sample analysis chip 100 rotates so that the second heater 332 is vertically aligned with the wax chamber 190.

Next, the heating unit 330 is moved by the driving unit 340 in a direction closer to the sample analysis chip 100, and finally the second heater 332 comes into contact with the portion where the wax chamber 190 is formed. .

Next, the second heater 332 is heated, and the solid wax stored in the wax chamber 190 is melted.

Next, the heating unit 330 moves in a direction away from the sample analysis chip 100 by the driving unit 340.

Next, the sample analysis chip 100 rotates and the melted wax (oil) is dispensed into the input channel 174.

Next, the heating unit 330 is moved by the driving unit 340 in a direction closer to the sample analysis chip 100, and finally the first heater 331 comes into contact with the portion where the reaction chamber 180 is formed. .

Next, the first heater 331 is heated to a temperature of 95°C -> 56°C -> 72°C, which is the PCR temperature cycle, according to a preset temperature cycle, and the analysis target in the reaction chamber 180 Amplifies matter. Here, temperature control of the first heater 331 may be performed by varying the direction and intensity of the current supplied to the Peltier element 333 connected to the first heater 331.

Next, the heating unit 330 moves in a direction away from the sample analysis chip 100 by the driving unit 340, and the sample analysis chip 100 rotates so that the reaction chamber 180 and the sensor unit 350 Sorted top and bottom.

Next, the sensor unit 350 radiates light of a specific wavelength toward the reaction chamber 180 and detects light (fluorescence) of a specific wavelength emitted from the reaction chamber 180. Here, the light detection method by the sensor unit 350 may be the point-by-point or moving scanning method described above.

Next, the calculation processing unit 360 uses the fluorescence intensity detected by the sensor unit 350 to determine whether a substance to be analyzed is present in each reaction chamber 180 or whether the sample is infected with a disease. Information on different primers stored in each reaction chamber 180 is stored in advance in the calculation processing unit 360, and the calculation processing unit 360 determines, for example, the reaction chamber 180 with the reference fluorescence intensity as the reference reaction chamber. And, by determining other reaction chambers based on the determined reference reaction chamber, it is possible to determine whether a substance to be analyzed is present in each reaction chamber or whether the sample is infected with a disease.

The heating process of the reaction chamber 180 by the first heater 331 and the light detection process by the sensor unit 350 may be performed alternately. For example, the first heater 331 may be used in one PCR temperature cycle. After heating the reaction chamber 180 to follow, a light detection process by the sensor unit 350 may be performed, followed by a heating process by the first heater 331, and then light detection by the sensor unit 350. The processes can be performed alternately. Through this, it is possible to measure how the fluorescence intensity of each reaction chamber 180 changes as the PCR cycle progresses. Through this, it is possible for the calculation processing unit 360 to easily determine whether the sample is infected with a disease. For each disease, a standard infection fluorescence intensity determined according to the number of standard PCR cycles is set, so it is possible to determine whether a sample is infected with a disease using the fluorescence intensity information detected in the reaction chamber 180 each time a PCR cycle is performed.

Verification experiment 1

An experiment was conducted to prove the superiority of the sample analysis chip 100 according to the present invention.

A yellow-colored solution is injected into the sample storage unit 110 and the cleaning liquid storage unit 130, and a purple-colored solution is injected into the cocktail storage unit 150 and the eluent storage unit 160, the process according to FIG. 10 An experiment was performed by repeatedly rotating and stopping the sample analysis chip 100.

As a result of the experiment, it was confirmed that only the purple-colored solution was input into the reaction chamber 180, and only the yellow-colored solution was input into the waste liquid chamber 171.

In other words, the sample analysis chip 100 according to the present invention confirmed that each solution was introduced only into the target configuration (waste liquid chamber or reaction chamber), and through this, the mixture to be analyzed (eluent + analyte target + cocktail) was confirmed to be introduced into the reaction chamber 180 (see FIG. 13).

Verification experiment 2

An experiment was performed to compare the capture efficiency of analyte substances with a conventionally used sample analysis chip.

The same amount of sample was input into the sample analysis chip shown in FIG. 23 and the sample analysis chip 100 according to the present invention, and PCR of the mixture introduced into the reaction chamber 180 through the process according to FIGS. 12A to 12L. was carried out. Then, the same amount of sample was added to the Qiagen kit, and PCR was performed on the extraction solution.

In addition, the capture efficiency of each silica bead was confirmed by varying the layers (double layer, triple layer, and quintuple layer) of the silica beads forming the glass filter installed in the capture passage 120 of the sample analysis chip 100 according to the present invention. .

As a result of the experiment, the conventional sample analysis chip shown in FIG. 23 showed a capture efficiency of about 82.01%, and the Kaizen kit showed a capture efficiency of 94.47%, while the sample analysis chip 100 according to the present invention showed a capture efficiency of about 82.01%. It was confirmed that a capture efficiency exceeding 99.94% was achieved, especially when a glass filter made of triple-layer silica beads was applied. Considering that in the case of the Kaizen kit, all processes such as sample input and washing solution input must be performed manually, it can be confirmed that the sample analysis chip 100 according to the present invention is not only convenient, but also achieves excellent capture efficiency of the analyte material. (see Figures 14 and 15).

Verification experiment 3

A verification experiment was conducted to verify the excellence of the analysis device 300 of the sample analysis chip 100 according to the present invention.

First, by comparing an embodiment (Thick disc) in which the thickness of the sample analysis chip 100 is constant throughout the radial direction and an embodiment (Thin disc) in which the thickness of the portion where the reaction chamber 180 is located is thinner than other portions, , a verification experiment was performed to determine whether the actual temperature control of the reaction chamber 180 could be performed according to the target temperature cycle.

As shown in FIG. 16, the verification experiment was performed by inserting a temperature sensor into the reaction chamber 180 to directly measure the temperature of the mixture introduced into the reaction chamber 180.

In addition, as shown in FIG. 17, the driving unit 340 is controlled differently for each of the thick disc and the thin disc, so that both the upper and lower parts of the sample analysis chip 100 are in contact with the first heater 331 and heated. Example (Top & Bottom), in which only the bottom of the sample analysis chip 100 is heated in contact with the first heater 331 (Bottom), only the top of the sample analysis chip 100 is heated by the first heater 331. The temperature of the reaction chamber 180 was measured by comparing the example (Top) heated in contact with the heater 331.

As a result of the experiment, as shown in FIGS. 18 to 20, the goal was to heat both the top and bottom (Top & Bottom) rather than heating only the top or bottom of the sample analysis chip 100. It was confirmed that the reaction chamber 180 was heated to a temperature similar to the temperature cycle (PCR temperature cycle). And, although there was no significant difference, it was confirmed that the reaction chamber 180 was heated to a temperature similar to the target temperature cycle for the thin disc rather than the thick disc.

Verification experiment 4

Using the sample analysis chip 100 according to the present invention, a verification experiment was performed to determine whether each of the plurality of reaction chambers 180 could be distinguished using the fluorescence intensity detected by the sensor unit 350.

After materials with different fluorescence intensities are accommodated in advance in each reaction chamber (Aref, A1 to A11), the sample analysis chip 100 is rotated at different speeds while the sensor unit 350 emits light of a specific wavelength. The reaction chamber 180 was irradiated, and the fluorescence signal generated therefrom was measured.

As a result of the experiment, it was demonstrated that the sensor unit 350 can detect the fluorescence signal generated in each reaction chamber 180 at the expected signal size regardless of the rotation speed of the sample analysis chip 100. It was demonstrated that the fluorescence signals generated in the reaction chamber 180 could be distinguished (see FIGS. 21 and 22).

Above, the present invention has been described with reference to the embodiments shown in the drawings so that those skilled in the art can easily understand and reproduce the present invention, but these are merely illustrative examples, and various modifications and equivalent alternatives can be made by those skilled in the art from the embodiments of the present invention. It will be appreciated that embodiments are possible. Therefore, the scope of protection of the present invention should be determined by the claims.

100: Chip for sample analysis
110: sample storage unit
120: Capture Passage
130: Washing liquid storage unit
140: delay unit
150: Cocktail storage unit
160: Eluent storage unit
170: connection chamber
171: Waste liquid chamber
172: collection chamber
174: Input channel
180: reaction chamber
190: wax storage unit
200: Cartridge
300: analysis device
310: housing
320: Chip installation part
330: heating unit
340: driving unit
350: sensor unit
360: Operation processing unit
370: output unit
400: Vibration prevention unit

Claims (15)

Sample storage unit 110;
a preprocessing unit located radially outside the sample storage unit 110 and preprocessing the sample introduced into the sample storage unit 110;
A reaction chamber 180 located radially outside the pretreatment unit and into which the sample pretreated by the pretreatment unit is introduced; and
A wax storage unit 190 that communicates with the reaction chamber 180 to be located radially inside the reaction chamber 180 and stores wax in a solid state therein.
A device for amplifying substances in the reaction chamber 180 and analyzing the sample in which a sample analysis chip 100 including a is installed,
a chip installation unit 320 on which the sample analysis chip 100 is seated and which includes a motor 322 that rotates the sample analysis chip 100;
It is installed on the upper body 336a and can be moved closer to or away from the reaction chamber 180 and the wax storage unit 190 while being aligned vertically with respect to the reaction chamber 180 and the wax storage unit 190. It is configured to heat the reaction chamber 180, and includes a first heater 331 that heats the reaction chamber 180, and a second heater 332 that is installed spaced apart from the first heater 331 and heats the wax storage unit 190. Heating unit 330; and
A driving unit 340 connected to the upper body 336a and the lower body 336b connected by an installation pillar to move the heating unit 330;
It includes a sensor unit 350 that is aligned up and down with respect to the reaction chamber 180 and is capable of detecting fluorescence generated in the reaction chamber 180,
A spring is installed on the outside of the installation pillar, and the heating unit 330 is installed on the upper and lower sides of the reaction chamber 180, respectively.
Device.
delete According to paragraph 1,
The heating unit 330 further includes a Peltier element 333 connected to the first heater 331.
Device.
delete According to paragraph 1,
The plurality of reaction chambers 180 are provided, and the plurality of reaction chambers 180 are formed along the circumferential direction of the sample analysis chip 100,
The first heater 331 is formed to extend along the circumferential direction,
Device.
According to clause 5,
The area of the first heater 331 is configured to cover at least all of the plurality of reaction chambers 180,
The area of the second heater 332 is configured to cover at least the wax storage unit 190,
Device.
According to clause 5,
The thickness of the portion of the sample analysis chip 100 where the reaction chamber 180 is formed is thinner than other portions,
Device.
According to paragraph 1,
It further includes an arithmetic processing unit 360 that determines whether a substance to be analyzed is present in the sample introduced into the sample storage unit 110 or whether the sample is infected with a disease based on the fluorescence intensity detected by the sensor unit 350. doing,
Device.
As an analysis method using the device according to paragraph 3,
(a) Injecting a sample into the sample storage unit 110;
(b) rotating the sample analysis chip in a preset direction at a preset rotation speed to preprocess the sample introduced in step (a), and introducing the preprocessed sample into the reaction chamber 180;
(c) moving the heating unit 330 in a direction closer to the sample analysis chip 100;
(d) heating the second heater 332 to a preset temperature and melting the wax stored in the wax storage unit 190;
(e) The sample analysis chip 100 rotates in a preset direction at a preset rotation speed, and the wax melted in step (d) is located in the input channel 174 radially inside the reaction chamber 180. ) and being dispensed into;
(f) heating the first heater 331 to follow a preset reaction temperature cycle; and
(g) detecting fluorescence generated in the reaction chamber 180 by the sensor unit 350; including,
Analysis method.
According to clause 9,
Step (a) includes the step of injecting the sample into the sample storage unit 110 by coupling the cartridge 200 storing the sample to the sample analysis chip 100,
After step (b) and before step (c), the sample analysis chip 100 is rotated so that the second heater 332 is vertically aligned with the wax storage unit 190,
In step (f), the first heater 331 performs polymerase chain reaction (Polymerase Chain Reaction, PCR), Reverse Transcription Polymerase Chain Reaction (RT-PCR), and Isothermal Amplification Reaction. ) further comprising the step of amplifying the sample introduced into the reaction chamber 180 by heating to follow the temperature cycle of one or more reactions,
Analysis method.
According to clause 9,
In step (f),
Further comprising the step of heating the first heater 331 connected to the Peltier element 333 to follow a preset reaction temperature cycle by controlling the intensity and direction of the current flowing through the Peltier element 333,
Analysis method.
According to clause 9,
After step (f), before step (g),
Further comprising: rotating the sample analysis chip 100 so that the sensor unit 350 and the reaction chamber 180 are aligned vertically,
Analysis method.
According to clause 9,
After step (g) above,
(h) determining, by the calculation processing unit 360, whether a substance to be analyzed in each reaction chamber 180 or whether the sample is infected with a disease based on the fluorescence intensity detected in step (g); doing,
Analysis method.
According to clause 13,
In step (h),
The calculation processing unit 360 uses the number of combination cases of the types of primers provided in the plurality of reaction chambers 180 and the fluorescence intensity of each reaction chamber 180 detected by the sensor unit 350, Further comprising determining whether a substance to be analyzed is present in the reaction chamber 180 or whether the sample is infected with a disease.
Analysis method.
According to clause 13,
A material having a reference fluorescence intensity is previously stored in at least one reaction chamber among the plurality of reaction chambers 180,
In step (h),
The calculation processing unit 360 determines the reaction chamber 180 that matches the reference fluorescence intensity among the fluorescence intensities of each reaction chamber 180 detected by the sensor unit 350 as the reference reaction chamber, and determines the reference reaction. Further comprising determining the position of the other reaction chamber 180 relative to the chamber,
Analysis method.