Fluid device, fluid delivery system, and treatment method of fluid containing metal nanoparticles

Abstract

To enable the separation of metal nanoparticles and the detection of a gene in a fluid where the metal nanoparticles are separated to be performed in different spaces.SOLUTION: A fluid device 101 includes: a gene amplification section 104 that stores a fluid containing metal nanoparticles and that heats the fluid using a photothermal effect of the metal nanoparticles to amplify a gene in the fluid; a metal nanoparticles separating section 105 that separates the metal nanoparticles from the fluid; and a gene detection section 106 that is a space different from the metal nanoparticles separating section 105 and that detects the gene in the fluid where the metal nanoparticles are separated in the metal nanoparticles separating section 105.SELECTED DRAWING: Figure 3A

Description

[Technical field]

[0001] The present disclosure relates to fluidic devices, fluid delivery systems, and methods for treating fluids containing metal nanoparticles. [Background technology]

[0002] In testing for infectious diseases such as respiratory infections and bloodstream infections, rapid detection of pathogens is required to speed up the initiation of appropriate treatment and prevent secondary infection. In addition, since there are a wide variety of pathogens that can cause infectious diseases, it is important to test for multiple pathogens simultaneously. Genetic testing based on the Polymerase Chain Reaction (PCR) method is quicker than conventional tests based on microbial culture, and has been actively researched in recent years.

[0003] In normal PCR, a microtube containing a reaction solution is placed in a heat block, and the temperature of the heat block is changed to repeat the steps of DNA denaturation, annealing, and extension (thermal cycle) to amplify the target DNA. At this time, the rate-limiting factor is not the heating and cooling time of the solution, but the heating and cooling time of the heat block, and the reaction usually takes about an hour.

[0004] A technology that utilizes the surface plasmon resonance of metal nanoparticles (plasmonic PCR) is being developed as a faster PCR method. When metal nanoparticles dispersed in a PCR solution are irradiated with light of a specific wavelength, each metal nanoparticle exhibits surface plasmon resonance, converting light energy into thermal energy and heating the solution around the metal nanoparticles. Because the metal nanoparticles, which are the heat source, are dispersed in the solution, the solution can be heated more uniformly and quickly than with conventional methods that use a heater to heat the solution from outside. The heating speed of metal nanoparticles is tens to hundreds of times faster than conventional methods, and gene amplification is completed in just a few minutes with plasmonic PCR.

[0005] One method for detecting multiple genes is to identify the type of gene by its position information. For example, in a microarray, probes are fixed at specific positions on a small substrate, and genes are detected by utilizing a hybridization reaction between the probes and the genes in the sample. By using the position information of the probes, it is possible to identify more than hundreds of thousands of genes.

[0006] Another example of multi-item gene detection using position information is real-time PCR. In real-time PCR, probes that bind to specific genes are typically modified with different fluorescent dyes, and the type of gene is identified from the wavelength of the detected fluorescent signal. The number of items that can be identified by fluorescence is limited by the resolution of the detector, and is currently limited to about four items.

[0007] Therefore, in order to simultaneously test more genes, the sample is divided into multiple wells and the genes are identified by the position information of the wells. In this case, the number of simultaneous test items is the product of the type of dye and the number of wells, making it possible to simultaneously test more genes.

[0008] Recently, technology has been developed to carry out chemical reactions such as nucleic acid amplification and gene detection in channels formed on small substrates (fluidic devices). Because chemical reactions take place in a tiny space, the amount of reagent used can be reduced. Other advantages include reduced risk of contamination because the reaction takes place in a closed channel, and the fact that fluidic devices can often be mass-produced, making it possible to carry out inexpensive testing. [Prior art documents] [Non-patent literature]

[0009] [Non-Patent Document 1] Nicole R., et al., Nat. Nanotechnol., 17, 984-992, 2022 [Non-Patent Document 2] Jiyong C., et al., Nat. Biomed. Eng., 4, 1159-1167, 2020 Summary of the Invention [Problem to be solved by the invention]

[0010] When performing plasmonic PCR and gene detection using metal nanoparticles in a fluid device, the inhibition of gene detection by the metal nanoparticles becomes a problem. For example, in the case of gene detection based on a fluorescent signal, it is known that the metal nanoparticles absorb the excitation light irradiated to the gene detection section for signal detection, and the resulting fluorescent signal decreases (Non-Patent Document 1). The decrease in the fluorescent signal can cause a decrease in the sensitivity of the genetic test, leading to false negatives. In addition, it is considered that the fine flow channels in the fluid device are clogged by the metal nanoparticles, which slows down the liquid delivery speed and causes an extension of the test time.

[0011] As a method for removing metal nanoparticles used in plasmonic PCR, Non-Patent Document 2 discloses a method using metal nanoparticles with a magnetic core. Plasmonic PCR is performed in a microtube using nanoparticles with a magnetic body covered with gold, and the nanoparticles are aggregated to the bottom of the microtube by a magnet. When excitation light is irradiated to the microtube, a fluorescent signal derived from genes is obtained from other than the bottom of the microtube. In this document, plasmonic PCR, nanoparticle removal, and gene detection are performed at the same location. With this method, gene identification information based on position information cannot be obtained, and the number of simultaneous tests is limited to about four items, which is the number that can be identified by fluorescence. On the other hand, it is necessary to simultaneously test about 30 items of genes for bloodstream infections and about five or more items of genes for respiratory infections.

[0012] Therefore, the present disclosure aims to provide a fluidic device, a liquid delivery system, and a method for processing a fluid containing metal nanoparticles that enable multi-item genetic detection by performing separation of metal nanoparticles and detection of genes in the fluid from which the metal nanoparticles have been separated in different spaces. [Means for solving the problem]

[0013] The fluidic device of the present disclosure comprises a gene amplification section that contains a fluid containing metal nanoparticles, heats the fluid using the photothermal effect of the metal nanoparticles, and amplifies genes in the fluid, a metal nanoparticle separation section that separates the metal nanoparticles from the fluid, and a gene detection section that is in a space separate from the metal nanoparticle separation section and detects genes in the fluid from which the metal nanoparticles have been separated in the metal nanoparticle separation section.

[0014] The liquid delivery system of the present disclosure also comprises a fluidic device including a gene amplification section that stores a fluid containing metal nanoparticles and heats the fluid by the photothermal effect of the metal nanoparticles to amplify genes in the fluid, a metal nanoparticle separation section that separates the metal nanoparticles from the fluid, and a gene detection section that is in a space separate from the metal nanoparticle separation section and detects genes in the fluid from which the metal nanoparticles have been separated in the metal nanoparticle separation section, and a liquid delivery device into which the fluidic device can be installed and that delivers the fluid from which the metal nanoparticles have been separated in the metal nanoparticle separation section from the metal nanoparticle separation section to the gene detection section.

[0015] In addition, the method of treating a fluid containing metal nanoparticles disclosed herein includes heating the fluid using the photothermal effect of the metal nanoparticles contained in the fluid to amplify genes in the fluid, separating the metal nanoparticles from the fluid, transporting the fluid from which the metal nanoparticles have been separated from a first space in which the metal nanoparticles are separated to a second space separate from the first space, and detecting the genes in the fluid in the second space. Effect of the Invention

[0016] According to the present disclosure, the separation of metal nanoparticles and the detection of genes in the fluid from which the metal nanoparticles have been separated are carried out in different spaces, thereby enabling multi-item gene detection. Problems, configurations and effects other than those described above will become apparent from the following description of the embodiments. [Brief description of the drawings]

[0017] [Figure 1] Configuration diagram of the fluidic device of Example 1 [Diagram 2] 1 is a cross-sectional view of a fluidic device according to a first embodiment. [Figure 3A] Device configuration diagram of the liquid delivery system of Example 1 [Figure 3B] Hardware block diagram of the control unit of the first embodiment [Figure 4] Operation flow of Example 1 [Diagram 5] Schematic diagram of the fluidic device in step (c) of FIG. 4. [Figure 6] Schematic diagram of the fluidic device in step (e) of FIG. 4. [Figure 7] Schematic diagram of a fluidic device in step (f) of FIG. 4. [Figure 8] FIG. 1 is a configuration diagram of a fluid device according to a first modified example of the first embodiment. [Figure 9] 1 is a cross-sectional view of a fluidic device according to a first modified example of the first embodiment. [Figure 10] FIG. 1 is a diagram showing the configuration of a liquid delivery system according to a first modified example of the first embodiment. [Figure 11] FIG. 1 is a configuration diagram of a fluid device according to a second modified example of the first embodiment. [Figure 12] 1 is a cross-sectional view of a fluidic device according to a second modified example of the first embodiment. [Figure 13] FIG. 1 is a diagram showing the configuration of a fluid system according to a second modified example of the first embodiment. [Figure 14] Configuration diagram of the fluidic device of Example 2 [Figure 15] 1 is a cross-sectional view of a fluidic device according to a second embodiment of the present invention; [Figure 16] Device configuration diagram of the liquid delivery system of Example 2 [Figure 17] Operation flow of Example 2 [Figure 18] Schematic diagram of a fluidic device in step (d) of FIG. 17. [Figure 19] Schematic diagram of a fluidic device in step (e) of FIG. 17. [Figure 20] Configuration diagram of the fluidic device of Example 3 [Figure 21] 3 is a cross-sectional view of the fluidic device of Example 3. [Figure 22]Device configuration diagram of the liquid delivery system of Example 3 [Figure 23] Operation flow of Example 3 [Figure 24] Schematic diagram of a fluidic device in step (d) of FIG. 23. [Diagram 25] Schematic diagram of a fluidic device in step (e) of FIG. 23. [Figure 26] Configuration diagram of the fluidic device of Example 4 [Figure 27] 4 is a cross-sectional view of the fluidic device of Example 4. [Figure 28] Device configuration diagram of the liquid delivery system of Example 4 [Figure 29] Operation flow of Example 4 [Diagram 30] Schematic diagram of a fluidic device in step (d) of FIG. 29. [Diagram 31] Schematic diagram of a fluidic device in step (e) of FIG. 29. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0018] The embodiments of the present invention will be described in detail with reference to the drawings. In the following embodiments, the components (including element steps, etc.) are not necessarily essential unless otherwise specified or considered to be obviously essential in principle.

[0019] Example 1 Hereinafter, a description will be given of Example 1. This example relates to a fluid device and a liquid delivery apparatus for carrying out gene amplification using metal nanoparticles, removal of metal nanoparticles using magnetic force, and multi-item gene detection based on a chromatography method.

[0020] Fig. 1 shows a plan view of a fluidic device 101, and Fig. 2 shows a cross-sectional view taken along line AA in Fig. 1. The fluidic device 101 includes a substrate 102, a sealant 109, and a solid phase carrier portion 110.

[0021] The substrate 102 is preferably made of a material that is stable to temperature and pressure changes, is not easily affected by the solution used in the chemical reaction, and has good moldability. Suitable materials for this purpose include, for example, cycloolefin polymer (COP), polycarbonate (PC), and acrylic resin (PMMA).

[0022] The substrate 102 includes a solution introduction section 103, a gene amplification section 104, a metal nanoparticle separation section 105, a gene detection section 106, a valve connection section 107, and a flow path 108. The gene amplification section 104 is capable of containing a fluid (solution) containing metal nanoparticles. The gene amplification section 104 heats the fluid by a photothermal effect using surface plasmon resonance of the metal nanoparticles, and amplifies genes in the fluid. The metal nanoparticle separation section 105 separates the metal nanoparticles from the fluid. The gene detection section 106 is a space different from the metal nanoparticle separation section 105, and detects genes in the fluid from which the metal nanoparticles have been separated in the metal nanoparticle separation section 105.

[0023] The metal nanoparticle separation section 105 and the gene detection section 106 are connected by a flow channel 108 .

[0024] The solution introduction section 103 and the valve connection section 107 are processed as through holes on the substrate 102, and open on the surface opposite to the surface to which the sealant 109 is attached. The gene amplification section 104, the metal nanoparticle separation section 105, the gene detection section 106, and the flow channel 108 are formed by cutting the substrate 102 and sealing it with the sealant 109.

[0025] 1 and 2, in this embodiment, the gene amplification unit 104 and the metal nanoparticle separation unit 105 are located in the same space on the substrate 102. However, the gene amplification unit 104 and the metal nanoparticle separation unit 105 may be located in different spaces on the substrate 102. In this case, the gene amplification unit 104 and the metal nanoparticle separation unit 105 need to be connected by a flow path.

[0026] The gene amplification section 104 and the metal nanoparticle separation section 105 are desirably shaped without corners in order to minimize residual liquid of the sample solution. An example of a desirable shape of the gene amplification section 104 and the metal nanoparticle separation section 105 is an ellipse as shown in Figures 1 and 2, but the shape is not limited thereto and may be, for example, a circle. Furthermore, the gene amplification section 104 and the metal nanoparticle separation section 105 may be in the form of a flow channel like the flow channel 108 shown in Figures 1 and 2.

[0027] The genetic detection unit 106 does not have to be a rectangular parallelepiped as shown in Figures 1 and 2, and may be, for example, a cylindrical shape. The genetic detection unit 106 may also be in the form of a flow channel like the flow channel 108 in Figures 1 and 2.

[0028] The flow channel 108 is not limited to a straight shape as shown in Figures 1 and 2, but may be, for example, serpentine. In the example of Figure 1, the flow channel 108 is a single flow channel with no branches, but it may be a flow channel branched into multiple branches. The cross-sectional shape of the flow channel 108 can be designed arbitrarily, and may be, for example, a square, rectangular, or circular shape.

[0029] The sealant 109 has a function of adhering to the substrate 102. The surface of the sealant 109 that adheres to the substrate 102 may have adhesive properties or may be pressure sensitive. The sealant 109 is preferably made of polyolefin (PO) or polypropylene (PP). Moreover, the sealant 109 is preferably a resin film having a thickness of about 0.1 mm.

[0030] The solid phase carrier part 110 is fixed to the fluidic device 101 in order to perform multi-item gene detection based on the chromatography method. A plurality of specific molecules that chemically react with the genes to be tested are fixed to the solid phase carrier part 110 based on position information. Examples of the specific molecules include nucleic acids, antigens, antibodies, and peptides.

[0031] The shape and material of the solid phase carrier part 110 are not particularly limited and any known material can be used as long as the solid phase carrier part 110 can diffuse the solution by capillary action. For example, the solid phase carrier part 110 can be a sheet or rod-shaped filter paper, silica gel, polyethersulfone, nitrocellulose, etc.

[0032] Next, a liquid delivery device capable of installing any of the fluidic devices shown in Figs. 1 and 2 or their modified examples will be described. As an example, Fig. 3A shows a liquid delivery device 201 (liquid delivery system 300) in which the fluidic device shown in Figs. 1 and 2 is installed. The liquid delivery system 300 includes a fluidic device 101 and a liquid delivery device 201. The liquid delivery device 201 can be used to deliver a solution within the fluidic device 101. In addition, the liquid delivery device 201 can supply light, air pressure, sensors, etc., required for carrying out gene amplification using metal nanoparticles, removal of metal nanoparticles using magnetic force, and multi-item gene detection within the fluidic device 101 to the fluidic device 101.

[0033] The liquid delivery device 201 includes a device holder 202 , a cover 203 , a pressure regulator 204 , a valve 205 , a temperature sensor 206 , a light source 207 , a control unit 208 , a sensor 209 , a signal detection unit 210 , and an external magnetic source 211 .

[0034] It is desirable to use a material having heat resistance and low thermal conductivity for the device holder 202. For example, PEEK (polyether ether ketone) or PC (polycarbonate) is preferable.

[0035] The cover 203 serves to fix the fluidic device 101 in a predetermined position, which is placed on the device holder 202. It is preferable that the material of the cover 203 is heat-resistant and has low thermal conductivity. For example, PC (polycarbonate) can be used.

[0036] The pressurizing / depressurizing device 204 is an example of a liquid delivery means, and is connected to the solution introduction section 103. The pressurizing / depressurizing device 204 delivers a solution by pressurizing or depressurizing the inside of the fluidic device 101 via the solution introduction section 103.

[0037] The valve 205 is, for example, an on-off valve. The valve 205 may be a directional valve, or a tube may be connected to the valve connection portion 107 and a directional valve may be installed on the tube. The valve 205 may also be a microvalve that can be installed in the fluidic device 101.

[0038] The temperature sensor 206 is used to measure the temperature of the solution in the gene amplification unit 104. The temperature sensor 206 is, for example, a non-contact infrared sensor. The temperature sensor 206 may be a thermocouple that contacts the solution in the gene amplification unit 104.

[0039] The light source 207 is used to irradiate the solution in the gene amplification unit 104 with light of a specific wavelength and induce plasmon resonance of the metal nanoparticles in the solution. The light source 207 is, for example, an LED or a laser. The specific wavelength is selected to be close to the wavelength of the absorption maximum (reflection minimum) associated with the surface plasmon resonance of the metal nanoparticles used. The wavelength of the absorption maximum (reflection minimum) associated with the surface plasmon resonance of the metal nanoparticles is, for example, between 500 nm and 1400 nm, and varies depending on the composition, shape, or size of the metal nanoparticles. In FIG. 3A, in order to improve the efficiency of irradiation by the light source 207, the cover 203 in the area located between the light source 207 and the gene amplification unit 104 is opened. However, the shape of the cover 203 is not limited to this, and may not be open, for example.

[0040] The control unit 208 is used to control the temperature sensor 206 and the light source 207. The control unit 208 controls the temperature sensor 206 and the light source 207, thereby making it possible to control the temperature of the solution in the gene amplification unit 104 to a desired temperature. An example of the control of the solution temperature in the gene amplification unit 104 by the control unit 208 is as follows.

[0041] When light source 207 irradiates light onto the solution in gene amplification unit 104 under the control of control unit 208, the light energy is converted into thermal energy by surface plasmon resonance of metal nanoparticles, and the solution is heated. Control unit 208 acquires the temperature of the solution measured by temperature sensor 206, and when it determines that the solution has risen to a target temperature, it stops the light irradiation by light source 207. When it determines that the temperature of the solution has fallen to the next target temperature, control unit 208 resumes the light irradiation by light source 207, and the solution is heated again. The above steps are repeated a predetermined number of times to achieve a thermal cycle of PCR (Polymerase Chain Reaction).

[0042] Here, the hardware configuration of the control unit 208 will be described with reference to FIG. 3B. The control unit 208 has a processor 281, a main memory unit 282, an auxiliary memory unit 283, and an interface 284. The processor 281 is a central processing unit (CPU), a graphics processing unit (GPU), a digital signal processor (DSP), an ASIC, or the like. The main memory unit 282 is a dynamic random access memory (DRAM) or the like, and is used as a working area for the processor 281. The auxiliary memory unit 283 is a hard disk drive (HDD), a solid state drive (SSD), or a combination thereof, or the like, and stores various programs and various data. The interface 284 is a device controller or the like that controls the operations of peripheral devices (the pressure increase / decrease device 204, the movement mechanism 212, the temperature sensor 206, and the light source 207) connected to the control unit 208.

[0043] For example, the auxiliary storage unit 283 stores a temperature control program for implementing the above-mentioned PCR thermal cycle. The processor 281 executes the temperature control program and controls the start and stop of light irradiation by the light source 207 while monitoring the temperature information output from the temperature sensor 206.

[0044] The auxiliary storage unit 283 also stores a process control program for processing the fluid in the fluid device 101. The processor 281 executes the process control program and controls the operation of the pressurizing / depressurizing device 204 to send the fluid containing metal nanoparticles from the metal nanoparticle separation unit 105 to the gene detection unit 106 via the flow path 108. The processor 281 also controls the operation of the moving mechanism 212, which will be described later, to bring the external magnetic source 211 close to the metal nanoparticle separation unit 105 when separating the metal nanoparticles. The processor 281 also controls the operation of the moving mechanism 212, which will be described later, to keep the external magnetic source 211 close to the metal nanoparticle separation unit 105 when sending the fluid.

[0045] When the external magnetic source 211 is an electromagnet, the processor 281 (energization control unit) controls the energization of the external magnetic source 211 (electromagnet).

[0046] The sensor 209 acquires information to observe the gene detection reaction in the gene detection unit 106. The signal obtained by the sensor 209 is sent to the signal detection unit 210. At least one of a liquid level detection sensor, a fluorescence detection sensor, and an image recognition sensor can be used as the sensor 209. For example, when a liquid level detection sensor is used, the position of the solution in the solid phase carrier unit 110 can be more appropriately controlled. Furthermore, when an image recognition sensor is used, position information of the gene detected in the solid phase carrier unit 110 can be acquired and the type of gene can be identified. In order to improve the efficiency of observation by the sensor 209, the cover 203 in the area located between the sensor 209 and the gene detection unit 106 may be opened.

[0047] The external magnetic source 211 is used to remove metal nanoparticles containing a magnetic material contained in the solution in the metal nanoparticle separation unit 105 by magnetic force. The external magnetic source 211 is, for example, a magnet such as a neodymium magnet or a ferrite magnet. When a magnet is used as the external magnetic source 211, it is desirable to provide a moving mechanism 212 that changes the distance between the gene amplification unit 104 or the metal nanoparticle separation unit 105 and the external magnetic source 211, as shown in FIG. 3A. This allows the external magnetic source 211 to be moved away from the gene amplification unit 104 during plasmonic PCR. By moving the external magnetic source 211 away from the gene amplification unit 104, the metal nanoparticles are dispersed in the PCR solution, and the solution can be uniformly heated by surface plasmon resonance of the metal nanoparticles.

[0048] The external magnetic source 211 can be, for example, an electromagnet. When an electromagnet is used as the external magnetic source 211, the metal nanoparticles can be dispersed during the plasmonic PCR by starting to energize the electromagnet after the plasmonic PCR is completed. This makes a magnet driving mechanism unnecessary, and the liquid delivery device 201 can be further simplified and made smaller.

[0049] In addition, a mirror finish that reflects light in the irradiation wavelength band may be applied to the surface of the external magnetic source 211 or between the gene amplifier 104 and the electromagnet. This allows light that is not absorbed by the nanoparticles and is transmitted back into the solution, where it can be irradiated onto the metal nanoparticles. As a result, it becomes possible to more efficiently use the energy of the irradiated light to heat the solution.

[0050] The liquid delivery device 201 is not limited to the form shown in Fig. 3A, but may be changed depending on the shape of the fluidic device 101, the gene detection method in the gene detection unit 106, and other circumstances. For example, in Fig. 3A, the temperature sensor 206, the light source 207, and the sensor 209 are arranged on the sealing material 109 side of the fluidic device 101, but they may be arranged on the opposite side of the sealing material 109. In this case, it is desirable to cut out any region of the device holding unit 202 so as not to interfere with the operation of the temperature sensor 206, the light source 207, and the sensor 209.

[0051] 3A. Furthermore, a filter for preventing genetic contamination may be provided between the pressure-compression device 204 or the valve 205 and the fluidic device 101. The filter may be provided inside the fluidic device 101.

[0052] Furthermore, the liquid delivery device 201 may include a cooling mechanism for cooling the gene amplification unit 104. The cooling mechanism of the gene amplification unit 104 can speed up the cooling of the solution in gene amplification, thereby speeding up plasmonic PCR. The cooling mechanism may be air-cooling using a fan, for example. Alternatively, the cooling mechanism may be a method of bringing an external heat source close to the gene amplification unit 104.

[0053] Fig. 4 shows an example of an operation flow of a genetic test using the liquid delivery device 201. This operation flow is for carrying out plasmonic PCR using metal nanoparticles, removal of the metal nanoparticles, and multi-item gene detection. Each operation step will be described below with reference to Fig. 4.

[0054] First, a solution to be used in plasmonic PCR using metal nanoparticles is introduced from solution introduction section 103, and the solution is placed in gene amplification section 104 (FIG. 4(a)). The method of introducing the solution into fluidic device 101 is not limited to this, and for example, when liquid delivery device 201 is a syringe pump, the solution may be introduced into a syringe and then delivered from the syringe to the fluidic device.

[0055] The solution used in this embodiment is composed of, for example, a mixture of a sample liquid to be tested, one or more pairs of primers, a heat-stable enzyme, four types of deoxyribonucleoside triphosphates (dATP, dCTP, dGTP, dTTP), and metal nanoparticles.

[0056] The metal nanoparticles used in this embodiment contain a magnetic material in order to be removed by magnetic force after plasmonic PCR. Metal nanoparticles containing a magnetic material may be obtained, for example, by covering a core of a magnetic material with a metal layer. Alternatively, the metal nanoparticles may be obtained by forming a material in which a metal and a magnetic material are mixed into nanoparticles.

[0057] The size and shape of the metal nanoparticles can be determined arbitrarily, but the long side or diameter is from 1 nm to 1000 nm, preferably from 2 nm to 500 nm.

[0058] The metal nanoparticles used in this embodiment preferably contain gold. The metal nanoparticles may also contain an 11th group element such as silver or copper. Examples of materials for metal nanoparticles that exhibit surface plasmon resonance include magnesium, aluminum, gallium, indium, ruthenium, and nickel. When used in plasmonic PCR, it is necessary to select the metals with care, taking into account the inhibition of gene amplification by these metals and the chemical resistance of the metals.

[0059] In addition, since the metal nanoparticles used in this embodiment are dispersed in a solution and the solution is uniformly heated, it is desirable to perform surface modification to prevent the particles from agglomerating together. Surface modification can be performed by, for example, silica coating or phosphine sulfonic acid modification, which imparts an electric charge to the surface of the metal nanoparticles so that the particles repel each other electrostatically.

[0060] Of the solutions used in this embodiment, some or all of the components other than the sample liquid to be tested may be held in the fluidic device in a liquid or solid state in advance. For example, all components other than the sample liquid may be held in the gene amplification unit 104 in advance as a freeze-dried reagent, and the freeze-dried reagent may be dissolved by the sample liquid.

[0061] Next, the fluidic device 101 is fixed to the liquid delivery device 201 (FIG. 4(b)). In FIG. 4, a PCR solution is introduced into the fluidic device 101 (FIG. 4(a)), and then the fluidic device 101 with the PCR solution introduced therein is fixed to the liquid delivery device 201 (FIG. 4(b)). However, after the fluidic device 101 is fixed (FIG. 4(b)), the PCR solution may be introduced into the fixed fluidic device 101 (FIG. 4(a)).

[0062] Next, plasmonic PCR using metal nanoparticles is carried out in gene amplification unit 104 (FIG. 4(c)). FIG. 5 shows a schematic diagram of the position of solution 301 (a solution used for PCR other than metal nanoparticles) and the state of metal nanoparticles 302 at this point. At the time of FIG. 4(c), no magnetic force from external magnetic source 211 is acting on metal nanoparticles 302, and metal nanoparticles 302 are dispersed in solution 301.

[0063] Plasmonic PCR can employ a method called two-step PCR, in which denaturation, annealing, and extension are repeated in two temperature ranges to perform PCR. The desired thermal cycle can be achieved by alternately changing the temperature of the solution between the denaturation temperature (e.g., 95°C) and the annealing and extension temperature (e.g., 60°C) using the control unit 208.

[0064] Next, the external magnetic source 211 (here, a magnet) is brought close to the fluidic device (FIG. 4(d)). This causes a magnetic force to be applied from the external magnetic source 211 to the metal nanoparticles 302, causing the metal nanoparticles 302 to aggregate on the bottom surface of the metal nanoparticle separation section 105 on the external magnetic source 211 side (FIG. 4(e)). FIG. 6 shows a schematic diagram of the position of the solution 301 and the state of the metal nanoparticles 302 at this point.

[0065] Next, the solution 301 is moved to the genetic detection unit 106 by the pressure increase / decrease device 204 (FIG. 4(f)). FIG. 7 shows a schematic diagram of the position of the solution 301 and the state of the metal nanoparticles 302 at this point. In the case of FIG. 4(f), the external magnetic source 211 is kept close to the fluidic device 101. This allows the solution 301 to be sent to the genetic detection unit 106 with the metal nanoparticles 302 captured in the metal nanoparticle separation unit 105.

[0066] Finally, multi-item gene detection (here, nucleic acid chromatography) is carried out in the gene detection unit 106 (FIG. 4(g)).

[0067] In this embodiment, the double-stranded DNA amplified by PCR may be decomposed into single-stranded DNA before being used for gene detection. For example, when the chromatography performed in the gene detection unit 106 is based on a hybridization reaction between DNAs, the efficiency of the hybridization reaction is expected to be improved by decomposing the double-stranded DNA into single-stranded DNA. Methods for decomposing the double-stranded DNA into single-stranded DNA include, for example, thermal denaturation. Other examples include a method using a nucleic acid denaturant such as urea or formamide, and dissociation using a sodium hydroxide solution. Also, it is possible to obtain single-stranded DNA by obtaining double-stranded DNA in which the end of one strand is modified with biotin by PCR, and contacting a double-stranded DNA solution with beads or columns having an avidin modification on the surface.

[0068] Furthermore, enzymatic digestion with a nuclease such as lambda exonuclease can be used, in which case single-stranded DNA can be obtained by modifying the primers used in PCR with a recognition site for the nuclease.

[0069] Another method for improving the efficiency of gene detection based on hybridization reaction is to use a stop primer during PCR. PCR is performed using a primer that has a sequence (tag sequence) complementary to the probe used for gene detection and a modification that stops DNA elongation, to obtain double-stranded DNA with a single-stranded tag sequence at its end. Since the hybridization reaction occurs between the probe and the single-stranded tag sequence, the efficiency of gene detection can be improved.

[0070] So far, the configuration, liquid delivery device, and operation flow of a fluid device that employs chromatography as an example of a multi-item gene detection method have been described.

[0071] (Modification 1 of the first embodiment) Next, as a modification of Example 1, the configuration of a fluid device and a liquid delivery device for performing multi-item gene detection using a method other than chromatography will be described. Note that the operation flow is not shown because only the content of FIG. 4(g) is different. Also, explanations of parts common to FIG. 1 to FIG. 3B may be omitted.

[0072] Another example of a method for detecting multiple genes is, for example, a method of performing real-time PCR in multiple wells having position information. A plan view of a fluidic device 101 in which the gene detection unit 106 performs real-time PCR is shown in FIG. 8, and a cross-sectional view taken along line A-A in FIG. 8 is shown in FIG.

[0073] 8 and 9 includes a plurality of wells 401 to which position information is assigned in the gene detection unit 106. Each well 401 holds in advance, in a solid or liquid state, a PCR primer for amplifying a specific gene corresponding to the position information. Each well 401 can also hold in advance, in a solid or liquid state, reagents other than the primers used in PCR.

[0074] FIG. 10 shows a liquid delivery device 201 in which the fluid device shown in FIG. 8 and FIG. 9 is installed. The liquid delivery device 201 shown in FIG. 10 includes an external heat source 402 and a temperature control unit 403 in addition to the liquid delivery device 201 shown in FIG. 3A. The external heat source 402 has a built-in thermocouple and heater, and is kept at a desired temperature by the temperature control unit 403. The gene detection unit 106 is heated by the external heat source 402. A thermal cycle is achieved in the well 401 of the gene detection unit 106 by the control of the temperature control unit 403, and multiple gene detection based on real-time PCR is performed. The hardware configuration of the temperature control unit 403 is the same as that of the control unit 208 shown in FIG. 3B, so a description thereof will be omitted.

[0075] The liquid delivery device 201 is not limited to the form shown in Fig. 10. For example, the temperature sensor 206, the light source 207, and the sensor 209 are arranged on the sealing material 109 side of the fluidic device 101 in Fig. 10, but they may be arranged on the opposite side of the sealing material 109. Furthermore, the external heat source 402 is arranged on the opposite side of the sealing material 109 of the fluidic device 101 in Fig. 10, but they may be arranged on the sealing material 109 side.

[0076] 10. Furthermore, a filter for preventing contamination of genes may be provided between the pressure-pressurizing device 204 and the valve 205 and the fluidic device 101. Furthermore, the liquid delivery device 201 may include a cooling mechanism for the gene amplification unit 104.

[0077] (Modification 2 of Example 1) Another example of a multi-item gene detection method is a method based on a microarray method. Fig. 11 shows a plan view of a fluidic device 101 in which a gene detection unit 106 is configured to perform gene detection based on a microarray method, and Fig. 12 shows a cross-sectional view taken along line A-A in Fig. 11.

[0078] The fluidic device 101 shown in Figs. 11 and 12 includes a plurality of probe spots 404 to which position information is assigned in the gene detection unit 106. Probes corresponding to genes to be detected are fixed to the probe spots 404. The gene detection unit 106 is not limited to a single straight channel without branches as shown in Figs. 11 and 12, but may be, for example, meandering or branched into multiple channels. The gene detection unit 106 does not have to be in the channel shape shown in Figs. 11 and 12, but may be, for example, a rectangular parallelepiped shape as shown in Figs. 1 and 2, and the probe spots 404 may be two-dimensionally arranged in the gene detection unit 106 having such a shape.

[0079] Fig. 13 shows a liquid delivery device 201 in which the fluidic device 101 shown in Fig. 11 and Fig. 12 is installed. The liquid delivery device 201 shown in Fig. 13 includes an external heat source 402, a temperature control unit 403, a cleaning liquid storage 405, and a directional switching valve 406 in addition to the components of the liquid delivery device 201 shown in Fig. 1.

[0080] The external heat source 402 has a built-in thermocouple and heater, and is maintained at a predetermined temperature by a temperature control unit 403. The gene detection unit 106 is heated by the external heat source 402. Under the control of the temperature control unit 403, the gene detection unit 106 is heated to a temperature suitable for a hybridization reaction (for example, 45° C., 50° C., 55° C., or 60° C.), and multiple gene detection based on the microarray method is performed.

[0081] The cleaning solution storage 405 and the directional valve 406 are provided for use in a cleaning step to suppress background light during multiple gene detection. In the case of a gene detection method based on a fluorescent signal, such as the microarray method, background light can be reduced by performing a cleaning step after the gene detection reaction. In this embodiment, the cleaning step is achieved by the following operations.

[0082] After the hybridization reaction, the solution used in the hybridization reaction is discharged from the gene detection unit 106. The solution may be discharged to the outside of the liquid delivery device 201 through the valve 205. Alternatively, the solution may be discharged to the outside of the liquid delivery device 201 through the pressurizing and depressurizing device 204. Alternatively, a waste liquid chamber may be provided in the fluidic device 101, and the solution may be discharged into this. In this case, the gene amplification unit 104 or the metal nanoparticle separation unit 105 may also serve as the waste liquid chamber.

[0083] Next, the directional switching valve 406 is switched to connect the pressurizing and depressurizing device 204 to the cleaning solution storage 405. The pressurizing and depressurizing device 204 is operated to send the cleaning solution in the cleaning solution storage 405 to the gene detection unit 106, and the cleaning process is performed. The cleaning solution storage 405 is not limited to the position shown in FIG. 13, and may be located inside the fluidic device 101, for example.

[0084] The liquid delivery device 201 is not limited to the form shown in Fig. 13. For example, the temperature sensor 206, the light source 207, and the sensor 209 are arranged on the sealing material 109 side of the fluidic device 101 in Fig. 13, but they may be arranged on the opposite side of the sealing material 109. Furthermore, the external heat source 402 is arranged on the opposite side of the sealing material 109 of the fluidic device 101 in Fig. 13, but they may be arranged on the sealing material 109 side.

[0085] 13. Furthermore, a filter for preventing contamination of genes may be provided between the pressure-pressurizing device 204 and the valve 205 and the fluidic device 101. Furthermore, the liquid delivery device 201 may include a cooling mechanism for the gene amplification unit 104.

[0086] The double-stranded DNA amplified by PCR may be decomposed into single-stranded DNA before being subjected to the hybridization reaction. The decomposition into single-stranded DNA may be performed before or after the removal of the metal nanoparticles.

[0087] (Modification 3 of Example 1) Yet another example of the technique for multi-item gene detection is a technique based on a bead array method. In this case, the gene detection unit 106 is provided with a plurality of types of beads having probes fixed on the surface. The beads may be arranged two-dimensionally on the substrate 102 of the gene detection unit 106, which is a rectangular parallelepiped or cylindrical shape. The beads may also be arranged linearly on the gene detection unit 106, which has one or more flow paths. In multi-item gene detection based on the bead array method, genes may be identified by the order in which the beads are arranged. Genes may also be identified by labeling the beads with different fluorescent or other light.

[0088] (Variation 4 of the First Embodiment) Yet another example of the method for detecting multiple genes is a method based on electrophoresis. In this case, the gene detection unit 106 is provided with an electrophoretic carrier and an electrolyte. The electrophoretic carrier may be a capillary having an ionic functional group such as silica on the inner wall, or may be a material having a molecular sieve effect such as polyacrylamide gel or agarose gel. The electrolyte is not particularly limited as long as it is an ionic solution having electrical conductivity, and any known electrolyte may be used. In detecting multiple genes based on electrophoresis, genes may be identified by the migration distance of electrophoresis. Genes may also be identified by the migration time.

[0089] (Method for treating fluid containing metal nanoparticles) Here, a method for treating a solution (a fluid containing metal nanoparticles) will be described. The method for treating a solution is as follows: Heating the solution using the photothermal effect of metal nanoparticles contained in the solution (thermal cycling the solution) and amplifying the genes in the solution. Separating metal nanoparticles from a solution using magnetic force (magnet 211 or electromagnet), filter 501, buffer 601, gold thin film 701, etc. Sending a solution in which the metal nanoparticles have been separated from a first space (metal nanoparticle separation section 105) in which the metal nanoparticles are separated to a second space (gene detection section 106) different from the first space; and detecting the gene in the fluid in the second space. For example, the above-mentioned gene amplification, separation of metal nanoparticles, and delivery of a solution are performed under the control of the control unit 208. Furthermore, gene detection is performed by the signal detection unit 210.

[0090] The method of isolating the metal nanoparticles includes the steps of: A magnet (external magnetic source 211) is brought close to the first space (metal nanoparticle separation section 105) to separate the magnetic material-containing metal nanoparticles from the solution; or and separating the metal nanoparticles containing the magnetic material from the solution by energizing an electromagnet (external magnetic source 211) provided in the vicinity of the first space.

[0091] The solution from which the metal nanoparticles have been separated is sent to the second space. This includes sending the solution from which the metal nanoparticles have been separated to the second space (gene detection section 106) while a magnet (external magnetic source 211) is brought into close proximity to the first space (metal nanoparticle separation section 105) or while an electromagnet (external magnetic source 211) is energized.

[0092] (Effects of Example 1) According to the fluidic device 101 of the first embodiment, plasmonic PCR using metal nanoparticles can be performed in the fluidic device 101. Then, in the metal nanoparticle separation section 105, the amplified genes and the metal nanoparticles can be separated, and the solution in which the metal nanoparticles have been separated can be sent to the gene detection section 106. In the first embodiment, in the gene detection section 106 in a separate space in which the metal nanoparticles have been separated, it becomes possible to perform multi-item gene detection based on position information. Therefore, rapid and multi-item gene testing can be performed without the gene detection being hindered by the metal nanoparticles.

[0093] Furthermore, by connecting the metal nanoparticle separation section 105 and the gene detection section 106 via a flow channel, separation of metal nanoparticles and gene detection can be carried out in separate spaces within the same device.

[0094] By providing the gene amplification section 104 and the metal nanoparticle separation section 105 in the same space, the fluidic device 101 can be made smaller.

[0095] Example 2 Example 2 will be described below. This example relates to a fluid device and a liquid delivery device for carrying out gene amplification using metal nanoparticles, removal of metal nanoparticles using a filter, and multi-item gene detection based on a chromatography method. Note that the description of parts common to Example 1 may be omitted.

[0096] Fig. 14 shows a plan view of the fluidic device 101, and Fig. 15 shows a cross-sectional view taken along line A-A in Fig. 14. This embodiment differs from Example 1 in that, with respect to the configuration shown in Fig. 1, the gene amplification section 104 and the metal nanoparticle separation section 105 are provided in different spaces on the fluidic device 101, and in that the metal nanoparticle separation section 105 is provided with a filter 501.

[0097] The filter 501 in the metal nanoparticle separation unit 105 is used to remove metal nanoparticles from a solution after plasmonic PCR using metal nanoparticles is performed in the gene amplification unit 104. The filter 501 is, for example, an ultrafiltration membrane. The pore size of the filter 501 is not particularly limited as long as it is smaller than the short side or diameter of the metal nanoparticles and is a size that allows DNA to pass through, and any known filter can be used. The solution from which the metal nanoparticles have been removed by passing through the filter 501 is sent to the gene detection unit 106, where multiple gene detection is performed.

[0098] The gene amplification unit 104, the metal nanoparticle separation unit 105, and the gene detection unit 106 may be arranged in any order and at any position on the fluidic device 101. As shown in Figures 14 and 15, they do not have to be arranged in this order: gene amplification unit 104, metal nanoparticle separation unit 105, and gene detection unit 106. For example, the gene amplification unit 104 may be located between the metal nanoparticle separation unit 105 and the gene detection unit 106.

[0099] Next, a liquid delivery device capable of installing any of the fluidic devices shown in Fig. 14 and Fig. 15 or their modified examples will be described. As an example, Fig. 16 shows a liquid delivery device 201 in which the fluidic device shown in Fig. 14 and Fig. 15 is installed. The configuration shown in Fig. 16 differs from Example 1 in that it does not include an external magnetic source 211.

[0100] The liquid delivery device 201 of the second embodiment can be used to deliver a solution within the fluidic device 101. The liquid delivery device 201 can also supply light, air pressure, sensors, and the like to the fluidic device 101, which are required to perform gene amplification using metal nanoparticles, removal of metal nanoparticles using a filter 501, and multi-item gene detection within the fluidic device 101.

[0101] Fig. 17 shows an example of an operational flow of genetic testing using the liquid delivery device 201. This operational flow is for carrying out plasmonic PCR using metal nanoparticles, removal of the metal nanoparticles, and multi-item gene detection. The process from introducing the solution into the fluidic device 101 to carrying out plasmonic PCR using metal nanoparticles (Figs. 17(a) to (c)) is the same as that of Example 1 (Figs. 4(a) to (c)).

[0102] After the plasmonic PCR is completed, the solution 301 is sent toward the metal nanoparticle separation section 105 by the pressure-pressurizing device 204 (FIG. 17(d)). FIG. 18 shows a schematic diagram of the position of the solution 301 and the state of the metal nanoparticles 302 at this point. At the point in time of FIG. 17(d), the metal nanoparticles 302 are dispersed in the solution 301.

[0103] By continuing to send the solution further, the metal nanoparticles 302 are removed by the filter 501 (FIG. 17(e)). FIG. 19 shows a schematic diagram of the position of the solution 301 and the state of the metal nanoparticles 302 at this point. The metal nanoparticles 302 cannot pass through the filter 501 and are captured on the surface or inside of the filter 501. Meanwhile, the solution 301 containing DNA passes through the filter 501.

[0104] By continuing the liquid transfer by the pressure regulator 204, the solution 301 from which the metal nanoparticles 302 have been removed moves to the gene detection unit 106 (FIG. 17(f)). Finally, the gene detection unit 106 performs multiple gene detection (here, chromatography) (FIG. 17(g)).

[0105] As described in Example 1, the double-stranded DNA amplified by PCR may be decomposed into single-stranded DNA and then subjected to gene detection. The decomposition into single-stranded DNA may be performed before or after the removal of the metal nanoparticles.

[0106] Example 3 Example 3 will be described below. This example relates to a fluidic device and a liquid delivery device for carrying out gene amplification using metal nanoparticles, metal nanoparticle removal using a buffer, and multi-item gene detection based on a chromatography method. Note that the description of parts common to Example 1 or Example 2 may be omitted.

[0107] Fig. 20 shows a plan view of the fluidic device 101, and Fig. 21 shows a cross-sectional view taken along line A-A in Fig. 20. This embodiment differs from Example 2 in that a buffer 601 is provided in the metal nanoparticle separation unit 105 instead of a filter 501, as compared to the configurations shown in Figs. 14 and 15.

[0108] The buffer 601 in the metal nanoparticle separation unit 105 is used to remove metal nanoparticles from the solution after carrying out plasmonic PCR using the metal nanoparticles in the gene amplification unit 104. It is desirable that the metal nanoparticles used in the plasmonic PCR are dispersed in the PCR solution so that the solution is uniformly heated. Therefore, in order to prevent the aggregation of the metal nanoparticles, it is common to apply a surface modification having a charge to the surface of the metal nanoparticles so that the particles electrostatically repel each other.

[0109] In this embodiment, a buffer 601 is used that changes the salt concentration of the solution to neutralize the electric charge on the surface of the metal nanoparticles with salt, coagulate the metal nanoparticles, and remove them from the solution. The buffer 601 is, for example, a solution containing sodium chloride. There are no particular limitations on the buffer 601 as long as it has the effect of neutralizing the electric charge on the surface of the metal nanoparticles, and any known buffer can be used.

[0110] Buffer 601 may be held in advance in a liquid state within metal nanoparticle separation section 105. Alternatively, buffer 601 may be held in advance in a solid state within metal nanoparticle separation section 105, in which case solid buffer 601 may be dissolved by the solution after PCR.

[0111] Furthermore, the buffer 601 does not have to be held in advance in the metal nanoparticle separation section 105. For example, a chamber for storing the buffer 601 in a liquid state may be provided inside or outside the fluidic device 101, and the buffer 601 may be delivered from the chamber to the metal nanoparticle separation section 105.

[0112] Next, a liquid delivery device capable of mounting any of the fluidic devices shown in Figures 20 and 21 or their modified examples will be described. As an example, Figure 22 shows a liquid delivery device 201 in which the fluidic device shown in Figures 20 and 21 is mounted.

[0113] The liquid delivery device 201 of the third embodiment can be used to deliver a solution within the fluidic device 101. The liquid delivery device 201 can also supply light, air pressure, sensors, and the like to the fluidic device 101, which are required for carrying out gene amplification using metal nanoparticles within the fluidic device 101, removal of metal nanoparticles using a buffer 601, and multi-item gene detection.

[0114] Fig. 23 shows an example of an operational flow of genetic testing using the liquid delivery device 201. This operational flow is for carrying out plasmonic PCR using metal nanoparticles, removal of metal nanoparticles, and multi-item gene detection. The process from the introduction of the solution into the fluidic device 101 to the delivery of the solution 301 to the metal nanoparticle separation section 105 by the pressure-pressurizing device 204 (Figs. 23(a) to (d)) is the same as that of Example 2 (Figs. 17(a) to (d)).

[0115] 17 and 18, when the pressure-pressurizing device 204 sends the solution 301 toward the metal nanoparticle separation section 105, the metal nanoparticles 302 are dispersed in the solution 301. Figure 24 shows a schematic diagram of the position of the solution 301 and the state of the metal nanoparticles 302 at this point.

[0116] Furthermore, by continuing the liquid transfer, the buffer 601 in the metal nanoparticle separation section 105 is mixed with the solution 301 and the metal nanoparticles 302. This cancels out the electric charges on the surfaces of the metal nanoparticles 302, causing the metal nanoparticles 302 to aggregate and precipitate (FIG. 23(e)). FIG. 25 shows a schematic diagram of the position of the solution 301 and the state of the metal nanoparticles 302 at this point.

[0117] The supernatant not containing metal nanoparticles is moved from the metal nanoparticle separation section 105 to the gene detection section 106 by the pressure increase / decrease device 204 (FIG. 23(f)).

[0118] Finally, multiple gene detection (here, chromatography) is carried out in the gene detection unit 106 (FIG. 23(g)).

[0119] As described in Example 1, the double-stranded DNA amplified by PCR may be decomposed into single-stranded DNA and then subjected to gene detection. The decomposition into single-stranded DNA may be performed before or after the removal of the metal nanoparticles.

[0120] Example 4 Example 4 will be described below. This example relates to a fluid device and a liquid delivery device for carrying out gene amplification using metal nanoparticles, removal of metal nanoparticles using optical concentration, and multi-item gene detection based on a chromatography method. Note that the description of parts common to Example 1 or Example 2 may be omitted.

[0121] Fig. 26 shows a plan view of fluidic device 101, and Fig. 27 shows a cross-sectional view taken along line A-A in Fig. 26. This embodiment differs from Examples 2 and 3 in that, compared to the configurations shown in Figs. 14 and 15 or Figs. 20 and 21, a gold thin film 701 is provided in metal nanoparticle separation section 105 instead of filter 501 or buffer 601.

[0122] The thin gold film 701 in the metal nanoparticle separation section 105 is used to remove metal nanoparticles from the solution after plasmonic PCR using metal nanoparticles is performed in the gene amplification section 104, by using a phenomenon called optical concentration.

[0123] When a gold thin film is irradiated with a laser, the plasmon resonance of the gold converts the light energy into heat energy. This photothermal effect causes the temperature of the solution near the gold thin film to rise rapidly, generating microbubbles in contact with the gold thin film and generating convection around the microbubbles. It is known that the particles in the solution stirred by the convection are concentrated around the microbubbles, a phenomenon known as optical concentration.

[0124] In this embodiment, in order to utilize the above phenomenon to concentrate and remove gold nanoparticles in the solution, a gold thin film 701 is provided in the metal nanoparticle separation section 105.

[0125] Next, a liquid delivery device capable of installing any of the fluidic devices shown in Fig. 26 and Fig. 27 or their modified examples will be described. As an example, Fig. 28 shows a liquid delivery device 201 in which the fluidic device shown in Fig. 26 and Fig. 27 is installed. The configuration shown in Fig. 28 differs from Examples 2 and 3 in that a gold thin film 701 is provided in the metal nanoparticle separation section 105 instead of a filter 501 or a buffer 601, and that a light source 702 is provided.

[0126] The liquid delivery device 201 of the fourth embodiment can be used to deliver a solution within the fluidic device 101. The liquid delivery device 201 can also supply light, air pressure, sensors, and the like to the fluidic device 101, which are required to perform gene amplification using metal nanoparticles, removal of metal nanoparticles using optical concentration, and multi-item gene detection within the fluidic device 101.

[0127] Light source 702 is used to irradiate metal nanoparticle separation section 105 with light of a specific wavelength and induce plasmon resonance in thin gold film 701. Light source 702 is, for example, a laser. The specific wavelength is selected to be close to the wavelength of the absorption maximum (reflection minimum) associated with the surface plasmon resonance of the thin gold film used.

[0128] Fig. 28 shows an example of an operational flow of genetic testing using the liquid delivery device 201. This operational flow is for carrying out plasmonic PCR using metal nanoparticles, removal of metal nanoparticles, and multi-item gene detection. The process from introducing the solution into the fluidic device 101 to delivery of the solution 301 to the metal nanoparticle separation section 105 by the pressure-pressurizing device 204 (Figs. 29(a)-(d)) is similar to that of Example 2 (Figs. 17(a)-(d)) and Example 3 (Figs. 23(a)-(d)).

[0129] As in Examples 2 and 3, when solution 301 is sent toward metal nanoparticle separation section 105 by pressure-pressurizing device 204, metal nanoparticles 302 are dispersed in solution 301. Figure 30 shows a schematic diagram of the position of solution 301 and the state of metal nanoparticles 302 at this point.

[0130] By continuing the liquid transfer, the solution 301 is placed on the thin gold film 701 in the metal nanoparticle separation section 105. The solution 301 and the thin gold film 701 are irradiated with light using the light source 702, and the metal nanoparticles 302 are concentrated by optical concentration (FIG. 29(e)). FIG. 31 shows a schematic diagram of the position of the solution 301 and the state of the metal nanoparticles 302 at this point.

[0131] The supernatant not containing metal nanoparticles is moved from the metal nanoparticle separation section 105 to the gene detection section 106 by the pressure increase / decrease device 204 (FIG. 29(f)). Finally, multi-item gene detection (here, chromatography) is carried out in the gene detection section 106 (FIG. 29(g)).

[0132] (Modification) The present invention is not limited to the above-mentioned embodiment, but includes various modified examples. The above-mentioned embodiment has been described in detail to clearly explain the present invention, and is not necessarily limited to those having all of the configurations described. It is also possible to replace a part of the configuration of one embodiment with the configuration of another embodiment, and it is also possible to add the configuration of another embodiment to the configuration of one embodiment. It is also possible to add, delete, or replace a part of the configuration of each embodiment with another configuration. [Explanation of symbols]

[0133] 101 Fluid Devices 102 Substrate 103 Solution introduction section 104 Gene Amplification Unit 105 Metal nanoparticle separation section 106 Gene Detection Unit 107 Valve connection 108 Channel 109 Sealing material 110 Solid phase carrier part 201 Liquid delivery device 202 Device holder 203 Cover 204 Pressure increase / decrease device 205 Valve 206 Temperature Sensor 207 Light source 208 Control Unit 209 Sensor 210 Signal detection unit 211 External magnetic force source 281 processor 282 Main memory 283 Auxiliary storage 284 Interface 301 Solution 300 Liquid Delivery System 302 Metal Nanoparticles 401 Well 402 External heat source 403 Temperature control unit 404 Probe Spot 405 Cleaning Solution Storage 406 Directional valve 501 Filter 601 Buffer 701 Gold thin film 702 Light source

Claims

1. a gene amplification unit that contains a fluid containing metal nanoparticles, heats the fluid by the photothermal effect of the metal nanoparticles, and amplifies genes in the fluid; a metal nanoparticle separation unit that separates the metal nanoparticles from the fluid; a gene detection unit that detects genes in the fluid from which the metal nanoparticles have been separated in the metal nanoparticle separation unit, the gene detection unit being a space different from the metal nanoparticle separation unit. A fluidic device comprising:

2. The metal nanoparticle separation unit and the gene detection unit are connected by a flow path. The fluidic device according to claim 1 .

3. The gene amplification section and the metal nanoparticle separation section are in the same space. The fluidic device according to claim 1 .

4. The metal nanoparticle separation unit has a filter that separates the amplified gene and the metal nanoparticles. The fluidic device according to claim 1 .

5. The metal nanoparticle separation unit has a buffer that changes the salt concentration of the fluid to aggregate the metal nanoparticles in the fluid. The fluidic device according to claim 1 .

6. The metal nanoparticle separation unit has a gold thin film that generates convection in the fluid by irradiating the fluid with light to concentrate the metal nanoparticles. The fluidic device according to claim 1 .

7. The gene detection unit detects multiple items of the amplified genes based on position information. The fluidic device according to claim 1 .

8. The gene detection unit has a plurality of wells to which position information is assigned and in which real-time PCR is performed. The fluidic device according to claim 7 .

9. The gene detection unit has a plurality of probe spots each having a probe corresponding to a gene to be detected and each probe spot is assigned with position information. The fluidic device according to claim 7 .

10. The metal nanoparticles include gold. The fluidic device according to claim 1 .

11. a fluidic device including: a gene amplification section that contains a fluid containing metal nanoparticles, heats the fluid by the photothermal effect of the metal nanoparticles, and amplifies genes in the fluid; a metal nanoparticle separation section that separates the metal nanoparticles from the fluid; and a gene detection section that is in a space different from the metal nanoparticle separation section and detects genes in the fluid from which the metal nanoparticles have been separated in the metal nanoparticle separation section; a fluid delivery device capable of installing the fluid device and delivering the fluid from which the metal nanoparticles have been separated in the metal nanoparticle separation section from the metal nanoparticle separation section to the gene detection section. A liquid delivery system comprising:

12. The metal nanoparticles include a magnetic material, a magnet and a moving mechanism for bringing the magnet close to the metal nanoparticle separation unit; or an electromagnet provided in the vicinity of the metal nanoparticle separation unit and a current control unit that controls current flow to the electromagnet. The liquid delivery system according to claim 11 .

13. heating the fluid by the photothermal effect of metal nanoparticles contained in the fluid, thereby amplifying genes in the fluid; separating the metal nanoparticles from the fluid; Sending the fluid from which the metal nanoparticles have been separated from a first space in which the metal nanoparticles are separated to a second space different from the first space; and detecting genes in the fluid in the second space. A method for treating a fluid containing metal nanoparticles.

14. Isolating the metal nanoparticles comprises: Bringing a magnet into proximity with the first space to separate the magnetically-containing metal nanoparticles from the fluid; or and separating the metal nanoparticles containing a magnetic material from the fluid by energizing an electromagnet provided in the vicinity of the first space. The method for treating a fluid containing metal nanoparticles according to claim 13.

15. The sending of the fluid from which the metal nanoparticles have been separated to the second space includes: and sending the fluid from which the metal nanoparticles have been separated to the second space while the magnet is in close proximity to the first space or while the electromagnet is energized. The method for treating a fluid containing metal nanoparticles according to claim 14.

Images