JP2020110092A - Gene detection system, gene detection chip, gene detection apparatus, gene amplification method, and gene amplification apparatus - Google Patents

Abstract

To enable the detection of genes by simple operation at a sample collection site.SOLUTION: The disclosed gene detection chip 101 has a core body 1 that detachably holds a puncture needle in a cavity 5, and a tube 3 spirally wound around the core body 1. By suctioning the upper end of a tube 3 with an intake tube 11, the sample collected by the puncture needle is sucked to the lower end of the tube 3 through a sample receiving port opened on the inner peripheral surface of the cavity 5. At the same time, a solution for performing PCR, which is held in a liquid reservoir 7, is sucked into the lower end of the tube 3. The sucked sample is sucked by the tube 3 while being mixed with the solution, and spirally moves in the tube 3. By heaters 15, 17, and 19 arranged around the spiral tube 3, the solution is heated to the thermal denaturation temperature in PCR, the primer binding temperature, and the extension temperature, in each round. The presence or absence of the amplification of a particular gene is detected by detecting the fluorescence from a fluorescent dye in the solution through a through hole 21.SELECTED DRAWING: Figure 1

Description

TECHNICAL FIELD The present invention relates to a gene detection system for detecting a specific gene from a sample collected from a subject, a gene detection chip, a gene detection device, a gene amplification method, and a gene amplification device suitable for use in the system.

From the body of a subject such as a patient, a tissue or cell in which a gene mutation to malignant tumor is expected is collected as a sample, and the DNA (deoxyribonucleic acid) possessed by the collected cell is derived from a malignant tumor mutant cell. By detecting whether or not it contains a specific gene such as the above gene, for example, a malignant tumor gene such as EGFR gene or K-ras gene, diagnosis of malignant tumor, selection of an appropriate treatment method, etc. are made. .. For example, a method has been adopted in which a sample is collected using a puncture needle (sensitizer), and the DNA of the collected sample is amplified by PCR (polymerase chain reaction) to detect it. A puncture needle is an instrument that allows cells to be directly collected from a cell mass suspected of having a cancer mutation or the like. The cells collected by the puncture needle are evaluated as to whether or not the EGFR gene and the K-ras gene, which are malignant tumor markers, can be detected. In the example of the EGFR gene, PCR is performed using two PCR primers (a forward primer and a reverse primer) designed to amplify a gene sequence that specifies the EGFR gene. The presence or absence of the EGFR gene is detected by. For the detection of DNA, fluorescence detection is performed using a fluorescent dye, which is called an intercalator and is incorporated into a double helix of DNA. The detection of DNA is not limited to fluorescence detection, but electrochemical detection, detection by light absorption, and the like are also possible. Patent Document 1 discloses an example of gene amplification and detection technology.

However, the conventional gene detection technique requires a complicated procedure to perform PCR, and therefore, it takes about 5 to 8 days before obtaining the result of the genetic test after collecting the tissue. Was needed. Therefore, while waiting for the result of the genetic test, the cells in the cell mass suspected of being a malignant tumor may be delayed in administration due to further amplification, or may be mutated to a cell different from the cell when the sample was collected. In some cases, the effect of medication treatment may not be obtained due to the above. In particular, young cancer patients have a rapidly progressing disease state, and the stage of malignant tumor may shift to the next stage by the time when the result of the genetic test is known after waiting time. As a result, the treatment based on the diagnosis result may be delayed.

Normal PCR is completed within 3 hours in principle. Up until now, genetic testing has been time-consuming because the process (pretreatment) of extracting DNA from the tissue after collecting the tissue has been complicated, so it is necessary to use specific biochemical experimenters or skilled testers. Only the technician was able to operate the pretreatment. Furthermore, since only specific laboratory technicians can operate, the fact that the test subjects are concentrated in a certain institution has also been a factor that delays the genetic test.

Re-table WO 2006/137551 Publication

Martin U. Kopp, Andrew J. de Mello, Andreas Manz, “Chemical Amplification: Continuous-Flow PCR on a Chip” SCIENCE VOL. 280, 15 MAY 1998

In order to alleviate the above-mentioned delay due to relying on a laboratory, PCR, which is a process of extracting a gene from a tissue collected as a sample and a process of amplifying the extracted gene, can be performed easily at the site of collection. The inventors of the present application have found that it must be effective to develop a method that can be executed by various operations. In developing this technique, the inventors of the present application proceeded with the study as follows.

As described above, gene detection is performed by extracting a specific gene from the collected cell mass and amplifying it by PCR. That is, extracting a gene from a cell mass is an important factor. In addition, since reagents and chemicals for extracting DNA from cell mass are expensive and many chemicals are harmful to the human body, it is desirable that the amount used is as small as possible. In addition, it is preferable that the sample collected as a sample be as small as possible in order to improve the reliability of the diagnosis by performing diagnosis by some methods. Further, it is necessary to prevent infection of a measurement person such as a laboratory technician at the time of measurement and also prevent contamination (contamination) between samples.

Through these studies, in order to carry out gene detection at an early stage, in addition to enabling gene detection on the spot where a sample is collected, (1) cell destruction and intracellular gene extraction are performed. , A sample pretreatment mechanism and a mechanism for amplifying extracted (that is, exposed) DNA by PCR are integrated by some method, and (2) an instrument in which these mechanisms are integrated The inventors of the present application have found that it is desirable to configure the chip as a size chip and (3) make the chip disposable (disposable). Further, the chip is desired to be capable of efficiently contacting a controlled heat source for melting (thermal denaturation) and extension of a gene in order to enable amplification of a specific gene by PCR. It was Note that in Non-Patent Document 1, like a normal microchemical chip, a meandering channel is provided in a small and flat chip, and PCR is performed in the process in which a solution containing DNA passes through this channel. The technology to do is disclosed.

The present invention has been made by the inventors of the present invention through such studies, and enables gene detection by a simple operation at the site of sample collection, thereby enabling gene detection in a short time. It is an object of the present invention to provide a gene detection system, a gene detection chip, a gene detection device, a gene amplification method, and a gene amplification device suitable for use in the system.

In order to achieve the above object, a first aspect of the present invention is a gene detection system for detecting a specific gene from a sample collected from a subject, which comprises a gene detection chip and the gene detection chip. And a chip accommodating portion that detachably accommodates.
The gene detection chip includes a core body having a cavity for inserting the sample, and a tube having a spiral portion spirally wound around the core body.
The core body is opened on the inner wall surface of the cavity, a sample receiving port for receiving the inserted sample, and a liquid reservoir for storing a solution for performing PCR for selectively amplifying the specific gene, The sample receiving port, the liquid reservoir, and a flow path that communicates the one end of the tube.
The chip accommodating part has a plurality of heaters arranged along the circumferential direction of the spiral part around the gene detection chip to be accommodated. The plurality of heaters heat the spiral portion to different temperatures for performing the PCR along the circumferential direction.
Then, in the gene detection system, the PCR is executed in the process of moving from the one end to the other end in the tube by being sucked by the suction pump sucking the other end of the tube and the suction pump. And a detector for detecting whether or not the solution containing the specimen contains the amplified specific gene.

The gene detection system with this configuration is used, for example, as follows. A solution for carrying out PCR that selectively amplifies a specific gene is stored in a liquid reservoir of the core of the gene detection chip. The solution is, for example, a surfactant for breaking out the cell membrane/nuclear membrane of the sample to expose DNA, two kinds of primers (forward primer and reverse primer) corresponding to a specific gene, DNA polymerase, and four kinds of DNA polymerases. When amplification is detected by detecting nucleotides and fluorescence, a fluorescent dye or the like having fluorescence by binding to DNA is included.

A specimen collected from a target site of the subject's body is inserted into the hollow portion of the core body. A suction pump is connected to the other end of the tube and the other end of the tube is aspirated. As a result, the inserted sample is sucked through the sample receiving port, and at the same time, the solution is sucked from the liquid reservoir, and the solution containing these enters the tube from one end of the tube. The solution entering the tube moves toward the other end of the tube. In the process of moving inside the tube, the solution moves spirally around the core body in the spiral part. In this process, the solution is repeatedly heated to a plurality of stages of temperature for performing PCR by a plurality of heaters arranged around the gene detection chip along the circumferential direction of the spiral portion. As a result, when the sample DNA contains a specific gene, the specific gene is amplified. The detector detects whether or not the solution in which the PCR has been performed contains the specific gene that has been amplified. If the solution contains the amplified specific gene, a gene detection result indicating that the sample DNA contains the specific gene can be obtained. As a result, it becomes possible to make a pathological diagnosis of the target site of the body of the subject, for example, whether or not the target cancer cells are present.

As described above, according to the gene detection system of the present configuration, it is possible to perform gene detection by a simple operation at the site of sample collection, thereby enabling gene detection in a short time. In particular, since the solution containing the sample moves three-dimensionally along the spiral part of the tube, unlike the conventional technique disclosed in Non-Patent Document 1, a kind of stirring effect is generated in the process of movement, and different components are mixed. Mixing is promoted. Thereby, amplification of a specific gene is efficiently performed. This also contributes to shortening the time required for gene detection. Since the gene detection chip can be attached to and detached from the chip housing portion, only the gene detection chip together with the puncture needle can be disposable (disposable). The gene detection system according to this configuration can also be used for the purpose of detecting whether or not a specific gene is contained in RNA (ribonucleic acid; ribonucleic acid) instead of DNA contained in a sample. For example, when RNA is transcribed into DNA by adding reverse transcriptase to the solution stored in the liquid reservoir, and the RNA contains a specific gene, the transcribed DNA can generate a specific gene. It is amplified by PCR.

A second aspect of the present invention is the gene detection system according to the first aspect, wherein the core body is obtained by inserting the puncture needle from which the specimen has been collected into the hollow portion to obtain the specimen. It can be inserted into the cavity.
According to this configuration, by inserting the puncture needle into the hollow portion of the core body, the sample collected by the puncture needle can be directly inserted into the hollow portion without transferring to another device.

A third aspect of the present invention is the gene detection system according to the first or second aspect, wherein the plurality of heaters are three-step reaction temperatures of thermal denaturation in PCR, primer binding, and extension. And at least three heaters for heating the tube.

According to this configuration, the three-step reaction of thermal denaturation, primer binding (that is, annealing), and extension proceeds efficiently.

A fourth aspect of the present invention is the gene detection system according to any one of the first to third aspects, wherein the liquid reservoir is divided into a first liquid reservoir and a second liquid reservoir. The passage connects the sample receiving port, the liquid reservoir, and the one end of the tube in the order from the sample receiving port, the first liquid reservoir, the second liquid reservoir to the one end of the pipe. ..

According to this configuration, a solution containing a surfactant is stored in the first liquid reservoir, and a solution containing, for example, two kinds of primers, DNA polymerase, and nucleotides is stored in the second liquid reservoir, thereby receiving the sample. The sample aspirated through the mouth is mixed with the surfactant in the first liquid reservoir, the cell membrane/nuclear membrane of the sample is efficiently destroyed, and the DNA contained in the cells of the sample is efficiently naked, and then the second sample is collected. In the liquid reservoir of, the mixing with other solution is efficiently performed. After that, the solution is aspirated into the tube. Therefore, the well mixed solution moves in the tube, so that the amplification of the specific gene is performed more efficiently.

A fifth aspect of the present invention is the gene detection system according to any one of the first to fourth aspects, wherein the core body includes the other end of the tube and the spiral portion. It further has a through hole which a portion between them crosses. Further, the detector includes an excitation light irradiation device that irradiates the excitation light of a fluorescent dye having fluorescence by binding to DNA toward the tube from one side of the through hole, and fluorescence emitted by the fluorescent dye. Is detected from the other side of the through hole.

According to this configuration, the solution stored in the liquid reservoir is made to contain a fluorescent dye, the solution in which PCR is performed is irradiated with excitation light, and the fluorescence generated by the fluorescent dye bound to the DNA is detected to amplify the solution. It is possible to detect whether or not the specific gene thus contained is contained in the solution.

According to a sixth aspect of the present invention, there is provided a gene detection chip, which is a tube having a core body having a cavity for inserting a sample and a spiral portion spirally wound around the core body. And are equipped with. Then, the core body is opened on the inner wall surface of the cavity, receives the inserted sample, a sample receiving port, and a liquid reservoir for storing a solution for performing PCR for selectively amplifying a specific gene. And a flow path that connects the sample receiving port, the liquid reservoir, and one end of the tube.

The gene detection chip according to this configuration is used in the gene detection system according to the first aspect of the present invention to perform gene detection by a simple operation at the site of sample collection as described in the first aspect of the present invention. It makes it possible to carry out, which enables gene detection in a short time.

A seventh aspect of the present invention is the gene detection chip according to the sixth aspect, wherein the core body is obtained by inserting the puncture needle from which the specimen is collected into the hollow portion. It can be inserted into the cavity.
According to this configuration, by inserting the puncture needle into the hollow portion of the core body, the sample collected by the puncture needle can be directly inserted into the hollow portion without transferring to another device.

An eighth aspect of the present invention is the gene detection chip according to the sixth or seventh aspect, wherein the liquid reservoir is divided into a first liquid reservoir and a second liquid reservoir. Connects the sample receiving port, the liquid reservoir, and the one end of the pipe in the order from the sample receiving port to the one end of the pipe through the first liquid reservoir, the second liquid reservoir.

By using the gene detection chip having this configuration in the gene detection system according to the fourth aspect of the present invention, as described in the fourth aspect of the present invention, it is possible to more efficiently amplify a specific gene. It will be possible.

A ninth aspect of the present invention is the gene detection chip according to any of the sixth to eighth aspects, wherein the core body is between the other end of the tube and the spiral portion. Further has a through hole traversed by.

By applying this configuration to the gene detection system according to the fifth aspect of the present invention, the gene detection result can be obtained by using fluorescence detection, as described in the fifth aspect of the present invention.

A tenth aspect of the present invention is a gene detection device for detecting a specific gene from a sample collected from a subject, the gene detection chip according to any one of the sixth to ninth aspects of the present invention A chip accommodating portion that removably accommodates is provided. The chip accommodating part has a plurality of heaters arranged along the circumferential direction of the spiral part around the gene detection chip to be accommodated. The plurality of heaters heat the spiral portion to different temperatures for performing the PCR along the circumferential direction. Then, the gene detection device executes the PCR in the process of moving from the one end to the other end in the tube by being sucked by the suction pump that sucks the other end of the tube and the suction pump. And a detector for detecting whether or not the solution containing the specimen contains the amplified specific gene.

In the gene detection device according to this configuration, the gene detection chip according to any one of the sixth to ninth aspects of the present invention is housed in the chip housing part to describe the first to fourth aspects of the invention. Works the same as.

An eleventh aspect of the present invention is the gene detection apparatus according to the tenth aspect, wherein the plurality of heaters are provided at the reaction temperatures of three stages of thermal denaturation in PCR, primer binding, and extension. It includes at least three heaters for heating the.

According to this configuration, the three-step reaction of thermal denaturation, primer binding (that is, annealing), and extension proceeds efficiently.

A twelfth aspect of the present invention is the gene detection apparatus according to the tenth or eleventh aspect, wherein the detector accommodates excitation light of a fluorescent dye having fluorescence by binding to DNA. Excitation light irradiation device that irradiates the tube from one side of the through hole of the gene detection chip according to the ninth aspect of the present invention, and fluorescence emitted by the fluorescent dye is detected from the other side of the through hole. And a fluorescence detection device that does.

The gene detection device according to this configuration functions in the same manner as described in the fifth aspect of the present invention by accommodating the gene detection chip according to the ninth aspect of the present invention in the chip accommodating portion.

A thirteenth aspect of the present invention is a gene amplification method for selectively amplifying a specific gene from DNA or RNA contained in a sample. In this gene amplification method, by preparing a tube having a spiral portion having a spiral shape and disposing a plurality of heaters along the circumferential direction of the spiral portion, the identification is performed along the circumferential direction. Heating the helix to different temperatures to perform PCR that selectively amplifies the gene for, and supplying one end of the tube with the sample and a solution for performing the PCR. And performing the PCR by moving the supplied solution containing the sample along the tube from the one end to the other end.

According to this configuration, the sample and the solution are spirally moved in the spiral portion in the process of moving toward the other end of the tube after being supplied to the one end of the tube. In this process, the solution containing the sample is repeatedly heated by the plurality of heaters arranged along the circumferential direction of the spiral portion to a plurality of stages of temperatures for performing PCR. As a result, when the sample DNA contains a specific gene, the specific gene is amplified by PCR. Since the solution moves three-dimensionally along the spiral part of the tube, unlike the prior art disclosed in Non-Patent Document 1, the solution moves three-dimensionally in the process of moving, which causes a stirring effect, Mixing of the ingredients is facilitated. Thereby, amplification of a specific gene is efficiently performed. When the gene amplification method according to this configuration is used in the gene detection system according to one embodiment of the present invention, it becomes possible to perform gene detection by a simple operation at the site of sample collection, which enables gene detection in a short time. It will be possible. When a specific gene is contained in RNA instead of DNA contained in a sample, RNA is transcribed into DNA by adding reverse transcriptase to a solution for performing PCR, and the transcribed DNA Specific gene is amplified by PCR.

A fourteenth aspect of the present invention is the method for amplifying a gene according to the thirteenth aspect, wherein the plurality of heaters are provided at the reaction temperatures of three stages of thermal denaturation in PCR, primer binding, and extension. It includes at least three heaters for heating the.

According to this configuration, the three-step reaction of thermal denaturation, primer binding (that is, annealing), and extension proceeds efficiently.

A fifteenth aspect of the present invention is a gene amplification device that selectively amplifies a specific gene from DNA or RNA contained in a sample. This gene amplification device is provided with a tube having a spiral part having a spiral shape and a PCR that is arranged along the circumferential direction of the spiral part and selectively amplifies the specific gene along the circumferential direction. A plurality of heaters for heating the spiral portion to different temperatures for performing, and a solution for performing the PCR containing the sample, which is supplied to one end of the tube, and the one end along the tube. To the other end of the solution moving means.

According to this configuration, the gene amplification method according to the thirteenth aspect of the present invention is performed by supplying a sample and a solution for performing PCR that selectively amplifies a specific gene to one end of the tube. can do. Since the solution moves three-dimensionally along the spiral part of the tube, unlike the prior art disclosed in Non-Patent Document 1, the solution moves three-dimensionally in the process of moving, which causes a stirring effect, Mixing of the ingredients is facilitated. Thereby, amplification of a specific gene is efficiently performed. By using the gene amplification device according to this configuration for the gene detection system according to one embodiment of the present invention, it becomes possible to perform gene detection by a simple operation at the site of sample collection, thereby enabling gene detection in a short time. It will be possible.

A sixteenth aspect of the present invention is the gene amplifying apparatus according to the fifteenth aspect, wherein the plurality of heaters have the temperature at three reaction temperatures of thermal denaturation, primer binding, and extension in the PCR. It includes at least three heaters for heating the.

According to this configuration, the three-step reaction of thermal denaturation, primer binding (that is, annealing), and extension proceeds efficiently.

INDUSTRIAL APPLICABILITY According to the present invention as described above, a gene detection system that enables gene detection by a simple operation at the site of sample collection, thereby enabling gene detection in a short time, and use in the system A gene detection chip, a gene detection device, a gene amplification method, and a gene amplification device suitable for the above are realized.

It is a perspective view which illustrates the composition of the gene detection chip by one embodiment of the present invention. It is a perspective sectional view of the lower part of the gene detection chip of FIG. It is a plane sectional view of the lower part of the gene detection chip of FIG. It is a figure which shows the structure of the lower part of the gene detection chip of FIG. 1, (a) is a front view, (b) is sectional drawing which expanded the cross-section structure along the flow path. FIG. 2 is a front cross-sectional view showing the gene detection chip of FIG. 1 together with a puncture needle held in its cavity. It is a perspective view which illustrates the chip|tip accommodation part which accommodates the gene detection chip of FIG. It is a figure which shows the structure of the chip accommodation part of FIG. 6, (a) is a front view, (b) is a BB sectional view, (c) is an AA sectional view. It is a perspective view which illustrates the gene detection apparatus with which the chip|tip accommodation part of FIG. 6 was mounted. It is a top view of the gene detection apparatus of FIG. 9 is a graph exemplifying an outline of temperature change of a solution in a tube in the gene detection device of FIG. 8. It is a sectional view which expands along a channel and illustrates a section structure of a lower part of a gene detection chip by another embodiment of the present invention. It is sectional drawing which expand|deploys and illustrates along the flow path the sectional structure of the lower part of the gene detection chip by another embodiment of this invention. It is a perspective sectional view of the lower part of the gene detection chip according to still another embodiment of the present invention.

FIG. 1 is a perspective view illustrating the configuration of a gene detection chip according to an embodiment of the present invention. This gene detection chip 101 has a core body 1 and a tube 3. The core body 1 is in the shape of a cylinder having a hollow portion 5 along the central axis 4, and has a diameter-expanded portion 6 having a larger diameter than other portions near the head. A puncture needle (not shown) can be inserted into the hollow portion 5. A tube 3 is spirally wound around the outer peripheral surface of the core body 1. A liquid reservoir 7 is formed in the lower part of the core 1 near the lower end of the tube 3. The liquid pool 7 is for pooling a solution for performing PCR that selectively amplifies a specific gene to be detected, for example, an EGFR gene or a K-ras gene that is a malignant tumor marker.

The solution to be retained in the liquid reservoir 7 is, for example, a surfactant for destroying the cell membrane/nuclear membrane of the sample to make DNA naked, and two kinds of primers (forward) corresponding to a specific gene to be detected. Primer and reverse primer), a DNA polymerase, four kinds of nucleotides (dNTP), and a fluorescent dye having fluorescence by binding to DNA. Base selectivity is not required for fluorescent dyes. Since the components of these solutions are well known in the art, detailed description thereof will be omitted.

The cavity 5, the liquid reservoir 7, and the lower end of the tube 3 are in communication with each other through a flow path (not shown) formed inside the core body 1. A hollow cylindrical intake pipe connection port 9 is fixed to the upper surface of the expanded diameter portion 6 of the core 1. The upper end of the pipe 3 communicates with the hollow portion 10 of the intake pipe connection port 9. By connecting the intake pipe 11 to the intake pipe connection port 9, the air in the pipe 3 can be sucked toward the upper end.

When the suction tube 11 sucks the air in the tube 3 while the puncture needle (not shown) that has collected the sample is held in the cavity 5, the sample is sucked from the puncture needle and at the same time, the solution is collected from the liquid reservoir 7. While being sucked and mixed, they are sucked from the lower end of the tube 3 into the inside of the tube 3 and move toward the upper end. The DNA contained in the sample is exposed due to the surfactant contained in the solution. The solution containing the sample moves spirally along the tube 3 in the process of moving. Heaters 15, 17, and 19 are arranged along the circumferential direction around the spiral portion 13 of the tube 3, which is a spirally wound portion. The heaters 15, 17, and 19 are set to reaction temperatures corresponding to the three-step reaction of thermal denaturation in PCR (ie, thermal denaturation of double strand into two single strands), primer binding (ie, annealing), and extension. , Heating the spiral part 13 of the tube 3. As a result, these reactions are performed on the solution moving in the spiral portion 13 every one revolution. As a result, if the specific gene is present in the DNA of the sample, the specific gene is amplified up to twice each time the solution makes one turn around the spiral portion 13.

A through hole 21 is formed in the expanded diameter portion 6 of the core body 1. A portion of the pipe 3 above the spiral portion 13 penetrates the enlarged diameter portion 6 so as to cross the through hole 21. Excitation light that excites the fluorescent dye in the solution is applied to the tube 3 through the through hole 21, and the fluorescence emitted by the fluorescent dye is detected through the through hole 21. As the number of amplified genes increases, the amount of fluorescent dye bound to the genes increases, and thus the intensity of emitted fluorescence increases. Therefore, it is possible to detect whether or not a specific gene is amplified, depending on whether or not fluorescence emission is detected from the solution that passes through the through hole 21 via the spiral portion 13. If the sample does not contain the specific gene, amplification does not occur, and if it does, the gene is amplified. Therefore, it is possible to detect whether or not the sample contains a specific gene by detecting the emission of fluorescence. In this way, the gene detection chip 101 makes it possible to detect on the spot whether or not a specific gene is contained in the sample collected by the puncture needle.

The spiral part 13 of the tube 3 is wound 35 times as an example. Thus, if it contains a particular gene in the sample, while the solution passes through the helical portion 13, a particular gene is amplified up to 2 ³⁵ times. When the pipe 3 exceeds the spiral portion 13, the pipe 3 gently rises, goes upward, and vertically penetrates the enlarged diameter portion 6. The through hole 21 formed in the enlarged diameter portion 6 intersects with the central axis 4 in a three-dimensional manner at a portion distant from the central axis 4 of the core body 1 in the radial direction. In the illustrated example, the central axis 23 of the through hole 21 is perpendicular to the central axis 4 of the core body 1.

In the core body 1, for example, the diameter of the main portion 25, which is a portion extending downward of the expanded diameter portion 21, is about 5 mm, the diameter of the expanded diameter portion 6 is about 10 mm, and the core body 1 protrudes above the expanded diameter portion 6. The diameter of the head portion 27, which is the portion to be formed, is about 5 mm, which is the same as the main portion 25. The core body 1 is made of plastic as an example. The tube 3 is, for example, a glass tube having an outer diameter of about 0.1 mm and an inner diameter of about 0.05 mm, a plastic tube, or a glass tube having an outer surface coated with plastic. As an example, a quartz glass tube whose outer surface is coated with polyimide can be used as the tube 3.

FIG. 2 is a perspective sectional view of the lower portion of the gene detection chip 101. Further, FIG. 3 is a plan sectional view of the lower portion of the gene detection chip 101. 2 and 3, the puncture needle 201 held in the cavity 5 of the core body 1 is also shown. Further, FIG. 4 is a diagram showing a configuration of a lower portion of the gene detection chip 101, (a) is a front view, and (b) is a sectional view in which a sectional structure is developed along a flow path. The tube 3 is arranged along a spiral groove 29 formed on the outer surface of the main portion 25 of the core body 1. In order to maintain good thermal contact between the heaters 15, 17, 19 and the tube 3, the groove 29 is provided outside the tube 3 so that the tube 3 protrudes somewhat outward from the outer surface of the main portion 25 of the core 1. It is formed shallower than the diameter. The lower end of the groove 29 communicates with a hole 31 formed near the outer surface of the core 1. The lower end of the tube 3 is inserted into this hole 31 and is fixed to the inner peripheral surface of the hole 31 with, for example, an adhesive. The outer surface of the core body 1 may be covered with the tube 3 by an adhesive tape having excellent heat resistance and thermal conductivity, for example, a polyimide adhesive tape so that the tube 3 does not bulge outward from the groove 29.

A liquid reservoir 7 is provided near the lower end of the pipe 3. In the illustrated example, the liquid reservoir 7 is formed as a recess opening on the outer surface of the core 1. A sample receiving port 33 for receiving the sample collected by the puncture needle 201 is opened on the inner peripheral surface of the hollow portion 5. A channel 35 extends radially outward from the sample receiving port 33. The liquid reservoir 7 communicates with the flow path 35 through the flow path 37. The hollow portion of the pipe 3 inserted into the hole 31 is further communicated with the flow passage 35 through the flow passage 39. In this way, the sample receiving port 33, the liquid reservoir 7, and the hollow portion of the tube 3 are in communication with each other through the flow paths 35, 37, 39.

A solution for amplifying a specific gene by PCR is stored in the liquid reservoir 7. Since the liquid reservoir 7 has a small size, the solution stored in the liquid reservoir 7 is retained as it is without flowing out. The size of the liquid reservoir 7 is, for example, about 0.3 mm in diameter. It is also possible to sell the gene detection chip 101 holding the solution in the liquid reservoir 7 to a medical institution or the like in a state of being sealed in a polyethylene bag or the like. After holding the solution in the liquid reservoir 7, the opening of the liquid reservoir 7 may be covered with a water-impermeable adhesive tape.

As illustrated in FIG. 2, the puncture needle 201 has a needle body (referred to as “stylet”) 203 and a tubular sheath (referred to as “cannula”) 205 that accommodates the needle body 203. doing. A holding portion 207 that holds the collected tissue is formed at the tip of the needle body 203. The needle body 203 can be advanced and retracted along the sheath 205, and accordingly, the holding portion 207 can be projected or retracted from the tip of the sheath 205. As shown, the cavity 5 is closed at the lower end. With the needle body 203 retracted into the sheath 205, the puncture needle 201 is inserted into the cavity 5 until the tip of the sheath 205 contacts the bottom surface of the cavity. Thereafter, the puncture needle 201 is operated so that the needle body 203 is exposed from the sheath 205, and if the needle body 203 is exposed, the tip of the needle body 203 is kept in contact with the bottom surface of the hollow portion 5, The sheath 205 is pulled up. As a result, the sample 41 held by the holder 207 can be exposed from the sheath 205 and face the sample receiving port 33. The position of the bottom surface of the hollow portion 5 is determined so that the sample 41 faces the sample receiving port 33.

In this state, when the upper end of the pipe 3 is sucked by the suction pipe 11 (see FIG. 1), the sample 41 is sucked into the flow path 35 from the sample receiving port 33, and at the same time, the solution is sucked from the liquid reservoir 7 into the flow path 35. These are mixed with each other while moving through the flow paths 35 and 39, or further when moving into the tube 3. The mixed solution or the solution being mixed moves in the tube 3 in the form of a lump (plug) having a certain width. The length of the plug is, for example, about 2 millimeters. Since the solution moves three-dimensionally along the spiral portion 13 of the pipe 3, mixing of different components is promoted by a kind of stirring effect in the process of movement.

FIG. 5 is a front sectional view showing the gene detection chip 101 together with the puncture needle 201 held in the cavity 5. Illustration of the detailed structure inside the gene detection chip 101 is omitted. The puncture needle 201 has a manual operation unit 245 attached to its head. The needle 203 can be exposed from the tip of the sheath 205 (see FIG. 2) by placing a finger on the finger rest 247 of the manual operation unit 245 and pushing the plunger 249 with the thumb. By further pressing the plunger 249, the needle body 203 can be instantly pulled back into the sheath 205 by the restoring force of the built-in spring. In this way, it is possible to collect a part of the tissue inside the body in the state of being housed in the sheath 205. The puncture needle 201 from which the specimen 41 has been collected in this manner is inserted into the cavity 5 as shown in the figure. By pressing the plunger 249 again, the needle body 203 can be exposed from the tip of the sheath 205. In order to pull up the sheath 205 while keeping the tip of the needle body 203 in contact with the bottom surface of the hollow portion 5 of the core body 1, it is preferable to push up the plunger 249 and pull up the manual operation portion 245.

FIG. 6 is a perspective view illustrating a chip housing portion that houses the gene detection chip 101. Further, FIG. 7 is a diagram showing a configuration of the same chip housing portion, (a) is a front view, (b) is a BB sectional view, and (c) is an AA sectional view. The gene detection chip 101 is attached to the center of the heaters 15, 17 and 19 arranged in a ring together with the puncture needle 201 (not shown). After that, the sample 41 is aligned with the sample receiving port 33 (see FIG. 2), and the suction pipe 11 sucks the upper end of the tube 3 (see FIG. 1) to start amplification of a specific gene by PCR. .. The chip housing 110 illustrated in FIGS. 6 and 7 holds the heaters 15, 17, and 19.

The chip accommodating section 110 has a tubular main body 43 having an open upper end, and a lid 45 detachably covering the upper end opening of the main body 43. Heaters 15, 17, and 19, an excitation light irradiation device 47, and a fluorescence detection device 49 are installed in the main body 43. The heaters 15, 17, and 19 are sequentially arranged along the circumferential direction so as to form a hollow portion 51 along which the gene detection chip 101 is detachably inserted along the central axis 53. The heaters 15, 17, and 19 are circumferentially separated from each other by a sheet-shaped or plate-shaped thermal insulator 20. In the illustrated example, the heaters 15, 17, and 19 have a fan-shaped planar cross-sectional shape. As an example, in each of the heaters 15, 17, and 19, a rod-shaped electric heater 57 is installed at the center of a heat conductor 55 made of aluminum. The main body 43 is provided with a vertically long slit 59 for allowing heat from the heaters 15, 17, 19 to escape to the outside air.

The heaters 15, 17, 19 contact the spiral portion 13 of the tube 3 of the gene detection chip 101 inserted in the hollow portion 51, and heat the spiral portion 13 to three temperatures. As an example, the three stages of temperature are set to a thermal denaturation temperature (eg, about 98° C.), a primer binding temperature (eg, about 62° C.), and an extension temperature (eg, about 75° C.) in PCR. Therefore, in the solution that passes through the spiral portion 13, the double-chain thermal modification, the primer binding, and the extension are performed every time the solution makes one round. As a result, when the sample contains a specific gene, the specific gene is amplified up to twice each time the solution makes one round. In the illustrated example, for simplification, the heaters 15, 17, and 19 are arranged with a uniform width in the circumferential direction, but the heaters 15, 17, and 19 are arranged in the circumferential direction in accordance with the length of time for holding the solution at each reaction temperature. The width of can be freely distributed.

An excitation light irradiation device 47 and a fluorescence detection device 49 are installed on the upper side wall of the main body 43. The excitation light irradiation device 47 has a light emitting element that emits excitation light, for example, a light emitting diode. The fluorescence detection device 49 has a fluorescence light receiving element that receives fluorescence, for example, a phototransistor. The excitation light emitted from the excitation light irradiation device 47 passes through the through hole 61 provided in the main body 43 and the through hole 21 of the gene detection chip 101, and is applied to the tube 3. The fluorescence emitted from the solution in the tube 3 passes through the through hole 21 of the gene detection chip 101 and the through hole 63 provided in the main body 43, and is detected by the fluorescence detection device 49. Although not shown, it is desirable that a blue filter and a condenser lens through which the excitation light passes be arranged in the through hole 61. Similarly, it is desirable that an orange filter and a condenser lens through which fluorescence passes are arranged in the through hole 63.

The lid 45 has a through hole 65 formed at the center thereof through which the gene detection chip 101 can be inserted. Therefore, it is possible to attach the gene detection chip 101 to the chip housing portion 110 with the lid 45 attached to the main body 43, and remove the gene detection chip 101 from the gene detection chip 101.

FIG. 8 is a perspective view illustrating a gene detection device equipped with the chip housing 110. FIG. 9 is a plan view of the gene detection device. The gene detection device 120 includes, for example, three chip housing portions 110 and a main body portion 121 that detachably holds these chip housing portions 110. Terminals (not shown) to which the power lines of the heaters 15, 17, and 19 are connected are provided on the bottom surface of the chip housing portion 110. These terminals are connected to a connector (not shown) provided on the main body 121 when the chip housing 110 is attached to the main body 121 of the gene detection device 120. One end of the electric wire 66 is detachably connected to the excitation light irradiation device 47 (not shown) through another connector (not shown), and is attached to the fluorescence detection device 49 through yet another connector (not shown). One end of the electric wire 67 is connected to. The other ends of the electric wires 66 and 67 are connected to the main body 121. One end of the intake pipe 11 is detachably connected to the intake pipe connection port 9 (see FIG. 1) of the gene detection chip 101 housed in the chip housing portion 110. The other end of the intake pipe 11 is connected to the main body 121.

The electric wires 66 and 67 are arranged in the main body 43 of the chip housing portion 110, and other terminals (not shown) that are lined up with the terminals to which the power supply lines of the heaters 15, 17, and 19 are connected to the electric wires 66 and 67. The ends may be connected. In this case, by mounting the chip housing 110 on the main body 121 of the gene detection device 120, the excitation light irradiation device 47 and the fluorescence detection device 49 are connected to an electronic circuit (not shown) built in the main body 121. Connected.

The main body part 121 contains an electronic circuit including a pre-programmed computer circuit, a power supply circuit, and a suction pump (all are not shown). A display screen 69 is installed on the upper surface of the main body 121. When the switch 71 is turned on, the gene detection device 120 starts its operation, and the heaters 15, 17, and 19 held by the respective chip housings 110 are turned on. As a result, the heaters 15, 17, and 19 are maintained at the predetermined predetermined temperature. At the same time, an operation menu is displayed on the screen 69. The gene detection chip 101 for detecting a specific gene is housed in, for example, the central chip housing portion 110, and the intake pipe 11 is connected to the intake pipe connection port 9 (see FIG. 1) of the gene detection chip 101. After that, on the operation menu displayed on the display screen 69, when the switch for operating the gene detection chip 101 accommodated in the central chip accommodating portion 110 is touched, the suction pump is started, and the suction pipe 11 causes the pipe 3 to move. The upper end is aspirated. Thereby, amplification of a specific gene by PCR is performed.

When the plug of the solution crosses the through hole 21 (see FIG. 6) of the gene detection chip 101, the suction pump stops. The time required from the start of the suction pump until the plug of the solution crosses the through hole 21 is fixed. Therefore, the computer circuit of the gene detection device 120 can be programmed so that the suction pump is stopped after a lapse of a fixed time.

If fluorescence is emitted from the solution irradiated with the excitation light from the excitation light irradiation device 47 when the plug of the solution crosses the through hole 21 of the gene detection chip 101, the fluorescence detection device 49 is emitted. Detected fluorescence. Whether or not fluorescence is detected is displayed on the display screen 69. In this way, the gene detection device 120 automatically detects whether or not a specific gene is contained in the sample. The time required for detection is about 15 minutes, which is remarkably shorter than that of the conventional method, which requires about 5 to 8 days. When the plug of the solution crosses the through hole 21 of the gene detection chip 101, the suction pump automatically stops, so that the intake pipe 11 is not contaminated with the solution. The gene detection chip 101 and the gene detection device 120 collectively correspond to an embodiment of the gene detection system of the present invention.

FIG. 10 is a graph exemplifying the temperature change of the solution passing through the spiral portion 13 of the tube 3 in the gene detection device 120 equipped with the gene detection chip 101. The plug of solution moves through tube 3 sufficiently slowly so that it is maintained at an approximately constant temperature as it passes through the regions of heaters 15, 17, and 19 as shown.

(Other embodiments)
FIG. 11 is a cross-sectional view illustrating a channel of a gene detection chip according to another embodiment of the present invention. Similar to FIG. 4B, FIG. 11 shows the sectional structure developed along the flow path. As shown, the liquid reservoir 7 (see FIG. 2) may be divided into two liquid reservoirs 81 and 83. For example, among the solutions for amplifying a specific gene by PCR, a solution containing a surfactant for destroying the cell membrane/nuclear membrane of the sample is held in the liquid reservoir 81 near the sample receiving port 33, and 2 A solution containing seed primers, DNA polymerase, 4 nucleotides, and a fluorescent dye can be held in a reservoir 83 near the tube 3. Thereby, from the sample taken in from the sample receiving port 33, the DNA is first made naked by the surfactant, and the solution containing the naked DNA is mixed with the solution containing two kinds of primers and the like. In this way, the process of taking out the DNA contained in the sample from the cells or cell nuclei and exposing it is efficiently performed by the solution of the surfactant before being diluted by being mixed with another solution.

FIG. 12 is a cross-sectional view illustrating a channel of a gene detection chip according to still another embodiment of the present invention. Similar to FIG. 11, FIG. 12 shows the sectional structure developed along the flow path. As illustrated, the divided liquid reservoirs 81 and 83 may be arranged in series in the flow path from the sample receiving port 33 to the tube 3. As a result, the liquid reservoirs 81 and 83 have not only the function of holding the solution but also the function of efficiently mixing the solution and the DNA of the sample. By holding the solution containing the surfactant for destroying the cell membrane/nuclear membrane of the sample in the liquid reservoir 81, the sample taken in from the sample receiving port 33 is first efficiently mixed with the surfactant by the liquid reservoir 81. Then, the DNA contained in the sample is stripped. By holding a solution containing two kinds of primers, a DNA polymerase, four kinds of nucleotides, and a fluorescent dye in the liquid reservoir 83, the naked DNA and these components in the solution are efficiently mixed. .. The mixed solution is sucked into the tube 3.

Further, as illustrated in FIG. 12, the diameter of the flow path 35 is enlarged toward the sample receiving opening 33 in order to easily take the sample 41 (see FIG. 2) into the flow path 35 through the sample receiving opening 33. Alternatively, the flow path 35 may be formed.

In the gene detection device 120, the fluorescence emitted by the fluorescent dye was used to detect the amplified specific gene. Other means can be used to detect the specific gene that has been amplified. For example, it is possible to detect whether or not the gene has been amplified by measuring the electric resistance value of the solution. If the method does not use fluorescence, it is not necessary to mix a fluorescent dye with a solution for amplifying a specific gene by PCR.

Although the example in which the three heaters 15, 17 and 19 are arranged in the chip accommodating portion 110 is shown, it is also possible to arrange two heaters. For example, in order to bring the temperature of a part of the spiral portion 13 of the tube 3 to the primer coupling temperature, natural cooling may be adopted without disposing a heater. It is also possible to arrange four or more heaters. For example, in order to accelerate the rise and fall of the temperature of the solution moving in the spiral portion 13, a heater for heating the spiral portion 13 to a temperature higher or lower than the target temperature is added to the region where the temperature transitions. May be. Alternatively, a large number of subdivided heaters are arranged along the circumferential direction of the spiral portion 13 so that the temperature curve can be freely input on the screen 69, and the spiral portion 13 can be input according to the input temperature curve. The current supplied to these heaters may be individually controlled by a computer circuit so as to heat the heaters.

In the gene detection chip 101, the hollow portion 5 of the core body 1 was formed as a hole into which the puncture needle 201 can be inserted. On the other hand, it is possible to form the cavity 5 so that any other sample collecting tool such as an endoscope capable of collecting the sample 41 can be inserted. Further, the cavity portion 5 may be formed in a form in which the sample 41 can be inserted by another device that transfers the sample 41 from the sample collecting tool, instead of directly inserting the sample collecting tool. Further, for example, the sample 41 may be directly inserted into the cavity 5 by inserting the sample 41 into the cavity 5 from a sample collection tool or another device. In the gene detection chip 102 illustrated in the perspective sectional view of FIG. 13, it is assumed that the sample 41 is directly put into the cavity 5. Therefore, the bottom surface of the cavity 5 is set at the same height as the bottom surface of the sample receiving port 33 so that the sample 41 can be easily guided to the tube 3 through the sample receiving port 33. The cavity 5 may be formed in a shape different from the shape along the central axis 4 (see FIG. 1). For example, the hollow portion 5 may be formed as a hole that is orthogonal to the central axis 4, or may be formed as a concave portion that opens to the side wall surface of the core body 1 similarly to the liquid reservoir 7. As described above, the cavity 5 is sufficient as long as the sample 41 can be inserted, whether the sample 41 is held by the sample collecting tool or other device or directly without being held by them.

In the gene detection chips 101 and 102, the core body 1 has a cylindrical shape, and the spiral portion 13 of the tube 3 has a vertical center axis and is formed in a spiral shape with a constant diameter. On the other hand, the tube 3 may be set in a spiral shape having different diameters depending on the height, or may be an elliptical shape instead of a circular shape, or a spiral shape tracing a polygonal contour, and the central axis of the spiral should be the vertical direction. Instead, it may be in another direction, for example, the horizontal direction.

In the gene detection chips 101 and 102, the spiral portion 13 of the tube 3 is wound around the core body 1. However, the core body 1 may be omitted to form the spiral portion 13. In this case, the sample 4l and the solution for executing PCR may be supplied to one end of the tube 3 by another means without the interposition of the core body 1. If the core body 1 is not provided, the heaters 15, 17, and 19 can be arranged inside the spiral portion 13. Further, the spiral diameter of the spiral portion 13 of the tube 3 and the inner diameter/outer diameter of the tube 3 itself may be configured to be considerably larger than those illustrated in FIG. 1 and the like, and in that case, PCR is also executed. It is possible. Further, the gene detection device 120 can be used not only for the specimen 41 but also for the purpose of simply amplifying a specific gene contained in some sample. In this case, means for detecting the amplification of a specific gene, such as the excitation light irradiation device 47 and the fluorescence detection device 49, may be omitted. Further, as a solution moving means for moving the solution from one end of the pipe 3 to the other end, instead of a suction pump that sucks the other end of the pipe 3, an air supply pump that supplies gas such as air from one end of the pipe 3 May be adopted.

DESCRIPTION OF SYMBOLS 1 core, 3 pipes, 4 central axis, 5 hollow part, 6 expanded diameter part, 7 liquid storage, 9 intake pipe connection port, 10 hollow part, 11 intake pipe, 13 spiral part, 15, 17, 19 heater, 20 thermal insulator, 21 through hole, 23 central axis, 25 main part, 27 head part, 29 groove, 31 hole, 33 sample receiving port, 35, 37, 39 channel, 41 sample, 43 body part, 45 lid, 47 Excitation Light Irradiation Device, 49 Fluorescence Detection Device, 51 Hollow Part, 53 Central Axis, 55 Heat Conductor, 57 Electric Heater, 59 Slit, 61, 63, 65 Through Hole, 66, 67 Electric Wire, 69 Display Screen, 71 Switch , 81, 83 liquid reservoir, 101, 102 gene detection chip, 110 chip accommodation part, 120 gene detection device, 121 main body part, 201 puncture needle, 203 needle body, 205 sheath, 207 holding part, 247 finger hook, 249 plunger.

Claims (16)

A gene detection system for detecting a specific gene from a sample collected from a subject,
A gene detection chip, and a chip accommodation portion that detachably accommodates the gene detection chip,
The gene detection chip,
A core body having a cavity for inserting the specimen,
A tube having a spiral part that is spirally wound around the core,
The core is
A sample receiving port that opens into the inner wall surface of the cavity and receives the inserted sample,
A reservoir for storing a solution for performing PCR for selectively amplifying the specific gene,
A flow path connecting the sample receiving port, the liquid reservoir, and one end of the tube,
The chip accommodating portion is
Around the gene detection chip to be housed, a plurality of heaters arranged along the circumferential direction of the spiral portion are provided, and the plurality of heaters are for performing the PCR along the circumferential direction. Heating the spiral to different temperatures,
The gene detection system,
A suction pump for sucking the other end of the tube,
The solution containing the sample, in which the PCR has been performed in the process of moving from the one end to the other end in the tube by being sucked by the suction pump, contains the amplified specific gene. A gene detection system further comprising a detector for detecting whether or not to perform. The gene detection system according to claim 1, wherein the core body is capable of inserting the specimen into the cavity by inserting a puncture needle that has collected the specimen into the cavity. The gene detection system according to claim 1 or 2, wherein the plurality of heaters includes at least three heaters that heat the tubes to three reaction temperatures of thermal denaturation, primer binding, and extension in the PCR. The liquid reservoir is divided into a first liquid reservoir and a second liquid reservoir,
The flow path connects the sample receiving port, the liquid reservoir, and the one end of the tube in the order from the sample receiving port to the one end of the pipe through the first liquid reservoir and the second liquid reservoir. The gene detection system according to any one of claims 1 to 3, which is communicated. The core further has a through hole that a portion of the tube between the other end and the spiral portion traverses,
The detector is
An excitation light irradiation device that irradiates excitation light of a fluorescent dye having fluorescence by binding to DNA toward the tube from one side of the through hole,
The fluorescence detection apparatus which detects the fluorescence which the said fluorescent dye emits from the other side of the said through-hole, The gene detection system in any one of Claim 1 to 4. A core body having a cavity for inserting a specimen,
A tube having a spiral portion that is spirally wound around the core,
The core is
A sample receiving port that opens into the inner wall surface of the cavity and receives the inserted sample,
A reservoir for storing a solution for performing PCR for selectively amplifying a specific gene,
A gene detection chip, comprising: a sample receiving port, the liquid reservoir, and a flow path communicating with one end of the tube. The gene detection chip according to claim 6, wherein the core body is capable of inserting the specimen into the cavity by inserting a puncture needle that has collected the specimen into the cavity. The liquid reservoir is divided into a first liquid reservoir and a second liquid reservoir,
The flow path connects the sample receiving port, the liquid reservoir, and the one end of the tube in the order from the sample receiving port to the one end of the pipe through the first liquid reservoir and the second liquid reservoir. The gene detection chip according to claim 6, which is communicated. The gene detection chip according to claim 6, wherein the core body further has a through hole that a portion of the tube between the other end and the spiral portion traverses. A gene detection device for detecting a specific gene from a sample collected from a subject,
A chip housing part detachably housing the gene detection chip according to any one of claims 6 to 9,
The chip accommodating portion is
Around the gene detection chip to be housed, a plurality of heaters arranged along the circumferential direction of the spiral portion are provided, and the plurality of heaters are for performing the PCR along the circumferential direction. Heating the spiral to different temperatures,
The gene detection device,
A suction pump for sucking the other end of the tube,
The solution containing the sample, in which the PCR was performed in the process of moving from the one end to the other end in the tube by being sucked by the suction pump, contains the amplified specific gene. A gene detection device further comprising: a detector for detecting whether or not to perform. The gene detection device according to claim 10, wherein the plurality of heaters include at least three heaters that heat the tubes to three reaction temperatures of thermal denaturation in PCR, primer binding, and extension. The detector is
An excitation light irradiating device for irradiating the excitation light of a fluorescent dye having fluorescence by binding to DNA toward the tube from one side of the through hole of the gene detection chip according to claim 9.
The gene detection device according to claim 10 or 11, further comprising: a fluorescence detection device that detects the fluorescence emitted by the fluorescent dye from the other side of the through hole. A gene amplification method for selectively amplifying a specific gene from DNA or RNA contained in a sample, comprising:
Preparing a tube having a spiral portion,
By arranging a plurality of heaters along the circumferential direction of the spiral portion, the spiral portion is moved to different temperatures along the circumferential direction for performing PCR that selectively amplifies the specific gene. Heating
Supplying one end of the tube with the sample and a solution for performing the PCR;
Performing the PCR by moving the supplied solution containing the sample along the tube from the one end to the other end. The gene amplification method according to claim 13, wherein the plurality of heaters include at least three heaters that heat the tubes to three reaction temperatures of thermal denaturation, primer binding, and extension in the PCR. A gene amplification device for selectively amplifying a specific gene from DNA or RNA contained in a sample, comprising:
A tube having a spiral part having a spiral shape,
A plurality of heaters arranged along the circumferential direction of the spiral portion and heating the spiral portion to different temperatures along the circumferential direction for performing PCR that selectively amplifies the specific gene. When,
A gene amplification device, comprising: a solution transfer unit that moves a solution containing the sample for performing the PCR, which is supplied to one end of the tube, from the one end to the other end along the tube. The gene amplification device according to claim 15, wherein the plurality of heaters include at least three heaters that heat the tubes to three reaction temperatures of thermal denaturation, primer binding, and extension in the PCR.