JP2012037363A - Detection method, kit for detection, and detection system of biologically-relevant molecules - Google Patents

Abstract

PROBLEM TO BE SOLVED: To prevent drying of a carrier such as a DNA chip to raise detection accuracy when automatic detection of biologically-relevant molecules is performed using the carrier, etc.SOLUTION: At least a part to which a probe is fixed of the carrier is immersed in a detection vessel in which drying prevention liquid is stored, the carrier immersed in the drying prevention liquid is irradiated with excitation light via the detection vessel, light from the biologically-relevant molecules emitted by irradiation with the excitation light is detected via the detection vessel. At this time, a kit for detection with a support member which supports the carrier to which the probe for detecting the biologically-relevant molecules is fixed, and the detection vessel in which the drying prevention liquid for immersing the carrier is stored is used. When the support member by which the carrier is supported is engaged into the detection vessel in which the drying prevention liquid is stored, at least the part to which the probe is fixed of the carrier is immersed in the drying prevention liquid.

Description

  The present invention relates to a method for detecting a biological molecule using a carrier on which a probe is immobilized, and particularly to a detection method, a detection kit, and a detection system for improving the accuracy of automatic detection.

In recent years, awareness of food safety has improved, and the importance of food inspection has increased.
Currently, there are two types of food inspections: the official inspection based on the Food Sanitation Law and the self-inspection that each company voluntarily performs for quality control. In the official inspection, the inspection by the culture medium method is generally performed, and usually about 3 to 5 days are required until the determination of the inspection result. For this reason, in the case of food with a short period until consumption, there was a problem that determination before shipment could not be performed.

Therefore, food manufacturers conduct original food inspections that can be determined earlier by using various methods such as an enzyme substrate method and an immunochromatography method. In such self-inspection, shortening of inspection time, simple operation, accurate inspection result, and low inspection cost are required, but current inspection methods have their merits and demerits, and all these performances are provided. There is no inspection method.
In this situation, food inspection technology that excels in specificity and reproducibility by applying genetic testing using a carrier such as a DNA chip to food inspection, and can perform multiple inspections quickly and easily, has attracted attention. Has been.

For example, Patent Document 1 discloses an invention relating to a method for analyzing a sample containing a fluorescently labeled biologically relevant molecule using a carrier on which the biologically relevant molecule is immobilized.
According to such a method for detecting a biological molecule, a plurality of tests can be simultaneously performed quickly and easily.

However, genetic testing using such a carrier such as a DNA chip requires a lot of work by an inspector and requires manipulation of a very small amount of liquid at a micro level. For this reason, there has been a problem that the influence by the manual work of the inspector is large and the inspection results are likely to vary.
Therefore, automatic inspection apparatuses have been developed in order to eliminate the intervention of inspectors and improve the reliability of inspection results.

For example, Patent Document 2 discloses an invention relating to a support member for a carrier that can be suitably used in an automatic inspection apparatus.
In this way, genetic testing using a carrier such as a DNA chip can be performed with an automatic testing device, so that the reliability of test results can be improved, and multiple tests can be performed more quickly and easily. It has become.

JP 2009-229398 A JP 2010-14620 A

Here, in order to accurately read the detection result of the biologically relevant molecule bound to the probe from a carrier such as a DNA chip, it is necessary to prevent drying of the surface on the side where the probe of the carrier is immobilized. As a general method for detecting bio-related molecules, there is a scanner system that collects and detects excitation light, and there is a fixed camera system that irradiates the excitation light and detects it by image analysis. The latter fixed camera system can detect all of the readings at once, but it is used for the interaction process such as hybridization to bind the probe and the biological molecule, and for washing the molecule that did not bind to the probe. This is because a solution containing a salt is used in the washing step, and therefore, when the carrier surface is dried, the salt is precipitated to prevent accurate detection.
Therefore, in the carrier of Patent Document 1, drying is prevented by covering the carrier with a glass cover after the cleaning step. Further, in Patent Document 2, it is possible to use a cleaning liquid containing a substance having deliquescence in the cleaning process and use it for detection without drying the carrier.

  However, the carrier of Patent Document 1 has a relatively short time to dry, and its shape is not suitable for automation. In the method of Patent Document 2, the influence of humidity and temperature is large, and humidity control in the detection process is necessary. In addition, the shape of the support member of the carrier is suitable for automation, but if the amount of liquid on the surface of the carrier is too large, the solution becomes a lens and the image is distorted. There was a need to do.

Therefore, the present inventors diligently researched a support member and a detection container having a shape that can sufficiently prevent the carrier from being dried and that are suitable for automation, and the carrier was attached to the detection container containing the drying prevention liquid. Fitting a support member, immersing at least the part of the carrier on which the probe is immobilized, and irradiating excitation light through the detection container to detect the light emitted from the biologically relevant molecule, a slight amount of dryness The present inventors have found that the biologically relevant molecules can be detected without drying the carrier with the prevention liquid, and the present invention has been completed.
That is, according to the present invention, at least a portion of a carrier on which a probe is immobilized is immersed in a detection container containing a drying prevention liquid, and the carrier is dried by irradiating the carrier with excitation light through the detection container. It is an object of the present invention to provide a detection method, a detection kit, and a detection system for a bio-related molecule that can detect the bio-related molecule without any problem.

  The method for detecting a bio-related molecule of the present invention is a method for detecting a bio-related molecule that binds to a probe immobilized on a carrier, wherein at least a portion of the carrier on which the probe is immobilized contains an anti-drying solution. Soaked in a detection container, irradiated the excitation light with a carrier immersed in the anti-drying liquid through the detection container, and the light from the biologically related molecules emitted by the excitation light irradiation through the detection container. As a method of detection.

  The biorelevant molecule detection kit of the present invention includes a support member that supports a carrier on which a probe for detecting a biorelevant molecule is immobilized, a detection container that contains an anti-drying solution that immerses the carrier, and It is set as the structure provided with.

  The biologically relevant molecule detection system of the present invention includes a support member that supports a carrier on which a probe that binds to the biologically relevant molecule is immobilized, and a solution containing the biologically relevant molecule is housed, and the carrier is immersed in this solution. A reaction container that performs a binding reaction between a biological molecule and a probe, a cleaning liquid, a cleaning container that immerses a carrier in the cleaning liquid to remove biological molecules that did not bind to the probe, and a drying prevention A detection container in which at least a portion of the carrier on which the probe is immobilized is immersed in the anti-drying liquid when the liquid is accommodated and the support member is supported, and the carrier is immersed in the anti-drying liquid An excitation light irradiator that irradiates excitation light via a detection container, a reader that detects light from biologically related molecules emitted by the irradiation of excitation light via a detection container, and excitation light or unnecessary Fluorescence filter that transmits fluorescence only prevents transmission of turbulent light, a support member, a reaction vessel, a washing container, the detection container, and the excitation light irradiator and reader, a configuration equipped with a transfer device for sequentially conveying.

  ADVANTAGE OF THE INVENTION According to this invention, when carrying out automatic detection of a bio-related molecule | numerator using carriers, such as a DNA chip, it becomes possible to prevent drying of a support | carrier and to improve detection accuracy.

It is a figure which shows the detection kit of embodiment of this invention. It is a perspective view of the detection kit of the embodiment of the present invention. It is a figure which shows the influence of drying of the support | carrier surface. It is a figure which shows the detection apparatus of embodiment of this invention. It is a figure which shows the autofluorescence test result (primary screening) for material selection of a detection container. It is a figure which shows the autofluorescence test result (secondary screening) for material selection of a detection container. It is a figure which shows the autofluorescence test result (thickness and the relationship of fluorescence intensity) for material selection of a detection container. It is a figure which shows the detection system of embodiment of this invention. It is a figure which shows the state of the coupling reaction process in the detection system of embodiment of this invention. It is a figure which shows the state of the washing | cleaning process in the detection system of embodiment of this invention. It is a figure which shows the state of the drying prevention liquid immersion process in the detection system of embodiment of this invention. It is a figure which shows the state of the detection process in the detection system of embodiment of this invention.

  Hereinafter, embodiments of a detection method, a detection kit, and a detection system for biologically relevant molecules of the present invention will be specifically described with reference to the drawings.

[Biologically relevant molecule detection kit]
The detection kit according to the present embodiment can be used for detection of biologically relevant molecules by the detection system according to the present embodiment, and prevents the carrier surface from drying to improve the detection accuracy of biologically relevant molecules bound to the probe. Is possible.
As shown in FIG. 1, the detection kit includes a support member 10 and a detection container 20. 2, (a) shows the support member 10 to which the carrier 30 is attached, (b) shows the detection container 20, and (c) shows a state in which these are fitted.

  The specific shape of the support member 10 is not particularly limited. For example, as shown in FIG. 2A, a main body part 11, a columnar carrier support part 12 protruding from the center of the lower surface of the main body part 11, and In addition, a configuration having the fitting portion 13 formed on the upper peripheral edge of the carrier support portion 12 can be adopted. The carrier support portion 12 of the support member 10 supports the carrier 30, and the fitting portion 13 is fitted with the detection container 20. Moreover, it can be set as the structure provided with the carrier attachment part in the lower end of the carrier support part 12. FIG.

  The method of attaching the carrier 30 to the carrier support portion 12 of the support member 10 is not particularly limited. For example, as shown in FIG. 2 (a), the carrier 30 is fitted into the center of the protrusion formed at the lower end of the carrier support portion 12. Examples thereof include a method or a method of attaching the carrier 30 to the lower end of the carrier support 12 with an adhesive.

  The material of the support member 10 is not particularly limited as long as it can support the carrier 30 and can be fitted to the detection container 20 and can immerse the lower end of the carrier support 12 to which the carrier 30 is attached in the drying preventing liquid 40. In order to suppress generation of unnecessary light (excitation light scattering or the like) in the detection device 50, the material of the support member 10 is preferably a material having as low autofluorescence as possible. Examples of materials having low autofluorescence include carbon, graphite, titanium black, aniline black, Ru, Mn, Ni, Cr, Fe, Co and Cu oxides, and carbides of Si, Ti, Ta, Zr and Cr. be able to. Synthetic resins such as polyethylene, polypropylene, polystyrene, phenol, melamine, urea, unsaturated polyester, epoxy, polyurethane, diallyl phthalate, silicon, polyimide, vinyl ester, vinyl chloride, ABS, fluorine, polyamide, polyacetal, polycarbonate, polyester, polyamide , May be mixed with at least one selected from the group consisting of synthetic rubber, elastomer, polylactic acid, polycaprolactone and polyglycol.

Although the specific shape of the detection container 20 is not particularly limited, for example, as shown in FIG. 2B, the detection container 20 includes a cylindrical body portion 22 having a bottom surface portion 21 and an open top portion. It can be set as the structure which has the fitting part 22a in the inner periphery of this opening part, and has a substantially quadrilateral flange in the outer periphery of an opening part. Further, it is preferable that the bottom surface has a uniform thickness and is a flat surface.
When the detection container 20 contains a predetermined amount of the drying prevention liquid 40 and is fitted to the support member 10 at the fitting portion 22a, the detection container 20 removes the carrier 30 attached to the lower end of the carrier support portion 12 of the support member 10 from the drying prevention liquid. 40 can be immersed.

  As a material for at least the bottom surface portion 21 of the detection container 20, a material that transmits excitation light and light from a biological molecule emitted by irradiation with the excitation light is used. Therefore, since the container material itself shines due to the excitation light, the material of the bottom surface portion 21 is preferably a material having low autofluorescence. In addition, since an image is taken through a container, it is preferable to have high transparency (haze) and low birefringence so that the image is not distorted. Moreover, it is preferable that the detection container 20 is a flexible material so that the fitting part 22a can be fitted with the support member 10 suitably. Examples of materials that satisfy these conditions include cycloolefin polymer (COP), polystyrene (PS), polypropylene (PP), polycarbonate (PC), polyethylene (PE), polyethylene terephthalate (PET), and polymethyl methacrylate resin ( PMMA), cyclic olefin copolymer (COC), polylactic acid (PLA) and the like.

  From the viewpoint of transmitting light, the thickness of the bottom surface portion 21 of the detection container 20 is preferably thinner as long as it is theoretically thin, and is preferably flat. Specifically, it is preferably 0.1 mm to 1.5 mm. When the detection container 20 is molded by injection molding, a container having a thickness of 0.3 mm to 0.7 mm can be used, and a container having a thickness of 0.3 to 0.5 mm is more preferable.

The carrier 30 is obtained by immobilizing a probe that binds to a biological molecule to be detected. For example, a DNA chip or a microarray is used. As the carrier 30, a commonly used carrier can be used.
As the probe, for example, a nucleic acid, a peptide, a protein, or the like can be used, and a probe that specifically binds to a biological molecule to be detected is used.
The biologically relevant molecule to be detected is not particularly limited as long as it binds to the probe. For example, nucleic acids such as DNA and RNA, peptides, proteins, sugar chains, cells, complexes thereof, and other molecules And the like.

The anti-drying liquid 40 is used to prevent the probe 30 surface of the carrier 30 from being dried. This is because when the surface of the carrier 30 is dried, salt is deposited on the surface, and as shown in FIG. 3, detection of fluorescence is hindered. A general buffer solution or the like can be used as the anti-drying solution 40. For example, a buffer solution such as phosphoric acid, acetic acid, citric acid, or boric acid can be used.
In addition, the thickness of the drying preventing liquid 40 between the carrier 30 and the bottom surface portion 21 in the detection container 20 may be 0.1 mm to 3 mm as long as the drying preventing liquid is in uniform contact with the chip surface. Is preferred.
It is preferable that the width of the carrier support portion 12 of the support member 10 be slightly smaller than the inner diameter of the detection container 20 because the amount of the anti-drying liquid can be reduced. Further, it is preferable to appropriately change the liquid thickness, the carrier support 12 and the inner diameter of the detection container so that the liquid is not sucked up due to the capillary phenomenon.

If the detection kit of the present embodiment is used, the carrier 30 is attached to the carrier support portion 12 of the support member 10, the drying prevention liquid 40 is accommodated in the detection container 20, and the support member 10 and the detection container 20 are fitted. Thus, the carrier 30 can be immersed in the drying preventing liquid 40. For this reason, it is possible to detect biologically relevant molecules without drying the carrier 30.
Further, according to the detection kit of the present embodiment, the carrier 30 can be prevented from being dried by a small amount of the anti-drying liquid 40. For example, when a 3 mm square DNA chip is immersed in the detection container 20 having a bottom area of 22.9 mm ² , the anti-drying solution 40 is about 10 μl and the liquid thickness is about 0.1 mm. Can be detected. This is because according to the detection kit of the present embodiment, the drying prevention liquid 40 is accommodated in the detection container 20 and the carrier 30 can be used in a form of being immersed in the drying prevention liquid 40 from above.

  According to the detection kit of the present embodiment, the surface of the carrier 30 can be suitably immersed by capillary action between the carrier 30 and the detection container 20 even if the amount of the drying preventing liquid 40 is smaller than 10 μl. .

[Detection device for bio-related molecules]
As shown in FIG. 4, the detection device 50 of the present embodiment includes an irradiator 51, a reader 52, and a control device 53. Excitation light 54 is irradiated from the irradiator 51 to the carrier 30 immersed in the anti-drying liquid 40 through the detection container 20. Thereby, the fluorescent substance bonded to the biologically relevant molecule emits light, and the fluorescence 55 from the biologically relevant molecule is input to the reader 52 via the detection container 20 and read.

The irradiator 51 is a device that irradiates the excitation light 54. For example, a laser transmitter using a filter for a UV lamp or a kinocene lamp can be used as the light source of the irradiator 51. When a UV lamp or a kinocene lamp is used, only light having a required wavelength is used as excitation light through a filter.
The reader 52 is a device that reads light from biologically related molecules bound to the probe of the carrier 30. When this biologically relevant molecule has a fluorescent label such as Cy5, the fluorescent label emits light when irradiated with the excitation light 54, and the fluorescence 55 is input to the reader 52 via the detection container 20.

The control device 53 inputs the reading result from the reader 52 to create image data, and / or recognizes the position of each spot from the reading result, removes the background, and outputs the fluorescence intensity of each spot.
Note that the image data reading method of the detection device 50 includes a camera fixing method that collectively reads image data of the entire surface on the side where the probe of the carrier 30 is fixed, and reading while scanning the surface of the carrier 30. And a scanner system that performs the above.

  Here, when the biologically relevant molecule is detected by fluorescence in this way, autofluorescence of the detection container 20 becomes a problem. The autofluorescence of the detection container 20 is fluorescence emitted from the material of the detection container 20 when the detection container 20 is irradiated with the excitation light 54. When autofluorescence increases, the fluorescence from biologically relevant molecules becomes relatively small, so the background increases and the intensity of the detected fluorescence 55 decreases. In addition, if there is a large autofluorescence due to the material on a spot that is not originally illuminated, there is a possibility that a false detection that the spot is illuminated as a result. That is, if the autofluorescence of the detection container 20 is large, the detection accuracy (reproducibility) decreases. Therefore, it is desirable to select a material with less autofluorescence as the material of the detection container 20.

Therefore, various materials were screened and suitable materials for the detection container 20 were selected. The results are shown in FIGS.
First, as shown in FIG. 5, polypropylene (PP), polyethylene (PE), polystyrene (PS), polyethylene terephthalate (PET), polymethyl methacrylate resin (PMMA), cyclic olefin copolymer (COC), cycloolefin polymer Each of (COP), polylactic acid (PLA), and glass was irradiated with excitation light with an exposure time of 5 seconds, and the detected fluorescence intensity was measured. The fluorescence intensity can be further reduced by reducing the thickness of the material. In the test of FIG. 5, materials of various thicknesses are used. The results shown in the figure indicate that polyethylene (PE), polystyrene (PS), and cycloolefin polymer (COP) exhibit a fluorescence intensity comparable to that of glass and are suitable.

  Next, polypropylene (PP), polyethylene (PE), polystyrene (PS), polycarbonate (PC), cycloolefin polymer (COP), and glass have the same thickness (about 1.6 mm) using the same molding method. A test plate was prepared, and the fluorescence intensity was measured with an exposure time of 5 seconds. As a result, as shown in FIG. 6, it was found that polystyrene (PS) and cycloolefin polymer (COP) showed the same fluorescence intensity as glass, and polypropylene (PP) also showed a relatively low fluorescence intensity.

  Next, for each of polypropylene (PP), polystyrene (PS), and cycloolefin polymer (COP), test plates having thicknesses of 0.3 mm, 0.5 mm, and 0.7 mm were prepared, and the exposure time was 5 times. The fluorescence intensity was measured as a second. As a result, as shown in FIG. 7, it was found that the fluorescence intensity of the cycloolefin polymer (COP) was the lowest, followed by polystyrene (PS) and polypropylene (PP) in this order. In particular, cycloolefin polymer (COP) and polystyrene (PS) exhibit autofluorescence intensity that is almost similar to glass or relatively low in the entire thickness range of 0.3 mm to 0.7 mm, and the detection container 20 It has been clarified that it can be suitably used as a material. In addition, when the thickness of polypropylene (PP) is 0.3 mm, the fluorescence intensity is relatively small, and it has been clarified that it can be suitably used as a material for the detection container 20.

[Biological molecule detection system]
As shown in FIG. 8, the detection system according to the present embodiment includes a detection device 50, a transport device 60, a reaction unit 70, a cleaning unit 80, an anti-drying liquid immersion unit 90, and a lifting device 100.
As the detection device 50 in the detection system, the detection device 50 of the present embodiment described above can be used. In the figure, the control device 53 is not shown.

  The transport device 60 is a device that grips the support member 10 with the chuck 61 and moves the support member 10 left and right along the transport path 62 with a motor (not shown). The transport device 60 sequentially transports the support member 10 to the reaction container 71 in the reaction unit 70, the cleaning container 81 in the cleaning unit 80, the detection container 20 in the drying preventing liquid immersion unit 90, and the detection device 50. The detection device 50 includes an excitation light irradiator 51 and a fluorescence reader 52, which may be integrated or separate.

  The reaction unit 70 is a device that performs a binding reaction between a probe immobilized on the carrier 30 and a biological molecule, and includes a reaction vessel 71 that contains a reaction solution 72 and a heater 73. The reaction solution 72 contains a sample such as a bio-related molecule to be detected. When the biological molecule to be detected is a nucleic acid, for example, a PCR (polymerase chain reaction) amplification product can be used as the reaction solution 72, and hybridization between the probe and the biological molecule in the reaction vessel 71 is possible. Is done. The heater 73 is a device for adjusting the temperature of the reaction solution 72 to an optimum temperature for bonding the probe and the biological molecule.

The washing unit 80 is a device for washing away a substance that has not been bound to the probe in the binding reaction between the probe and the biological molecule by the reaction unit 70 and removing it from the surface of the carrier 30. The cleaning unit 80 includes a plurality of cleaning containers 81 each containing a cleaning liquid 82, and the carrier 30 is sequentially immersed in the cleaning liquid 82 of each cleaning container to swing at least one of the transport device 60 and the cleaning container 81. As a result, the carrier 30 is washed.
Although the oscillating device is not shown in the figure, the conveying device 60 is provided with a drive unit that moves the support member 10 up and down slightly, moves it horizontally, or moves the cleaning container 81 up and down. It is possible to employ a device provided with a drive unit that can be moved in the lateral direction, a heater that is attached to adjust the temperature in the cleaning unit, and the like.

The cleaning liquid 82 may be different for each cleaning container 81 or the same one,
For example, SSC / SDS, SSC, NaAC, MgCl ₂ or the like can be used.
By providing a plurality of washing containers 81 in this manner, components that have not been combined with the probe attached to the carrier 30 can be reliably removed.

  The material of the reaction vessel 71 and the washing vessel 81 is not particularly limited. For example, polyethylene, polypropylene, polystyrene, polyvinyl chloride, polyvinyl acrylate, ABS resin, acrylic resin, polymethyl methacrylate resin phenol, polyamide, polyacetate Synthetic resins such as polycarbonate, polybutylene terephthalate, polyethylene terephthalate, and cyclic polyolefin, synthetic rubbers and elastomers, polylactic acid, polycaprolactone, polyglycolic acid, and the like can be used. These may be used alone or in admixture of two or more.

  The anti-drying liquid immersion unit 90 removably supports the detection container 20 containing the anti-drying liquid 40, the support member 10 is fitted to the detection container 20, and the carrier 30 attached to the support member 10 is dried. It is an apparatus for immersing in the prevention liquid 40.

  The elevating device 100 includes an elevating platform 101 and elevates the elevating platform 101 by a cylinder (not shown). A reaction unit 70, a cleaning unit 80, and an anti-drying liquid immersion unit 90 are placed on the lifting platform 101. Here, the raising / lowering of the reaction part 70, the washing | cleaning part 80, and the drying prevention liquid immersion part 90 may provide a raising / lowering apparatus independent to each. It is only necessary to be able to control the relative heights of the support member 10 that supports the carrier 30, the reaction unit 70, the cleaning unit 80, and the drying prevention liquid immersion unit 90, and thus the reaction unit 70, the cleaning unit 80, and the drying unit It is also possible to raise and lower the transport device 60 that transports the support member 10 by fixing the prevention liquid immersion portion 90.

[Detection method of bio-related molecules]
The detection of the bio-related molecule of this embodiment can be performed by the procedure shown in FIGS.
First, as shown in FIG. 9, the support member 10 is gripped by the transport device 60, moved in the horizontal direction, and the elevator 101 is raised in a state where the support member 10 is positioned directly above the reaction unit 70, so that the carrier 30 is moved into the reaction vessel. It is immersed in the reaction liquid 72 in 71 for a fixed time. The temperature of the reaction solution 72 is adjusted by the heater 73 to an optimum temperature for the binding between the probe on the carrier 30 and the biological related molecule to be detected. As a result, a binding reaction between the probe and the biological molecule is performed.
When the binding reaction step is completed, the lifting platform 101 is lowered, and the support member 10 is moved directly above the first cleaning container 81 in the cleaning unit 80 by the transport device 60.

Next, as shown in FIG. 10, in the state where the support member 10 is positioned directly above the first cleaning container in the cleaning unit 80, the lifting platform 101 is raised to move the carrier 30 to the first cleaning container. It is immersed in the cleaning liquid 82 inside. And the support | carrier 30 is rock | fluctuated by the conveying apparatus 60, and the support | carrier 30 is wash | cleaned.
Similarly, the support member 10 is sequentially moved from the first cleaning container to the fifth cleaning container, and the carrier 30 is sufficiently cleaned.
When the cleaning process of the carrier 30 is completed, the elevator 101 is lowered and the support member 10 is moved directly above the drying preventing liquid immersion unit 90 by the transport device 60.

  Next, as shown in FIG. 11, with the support member 10 positioned right above the drying prevention liquid immersion unit 90, the lift 101 is moved to a position where the support member 10 and the detection container 20 are fitted. Raise. Then, the support member 10 and the detection container 20 are fitted, and the carrier 30 is immersed in the anti-drying liquid 40 in the detection container 20. Next, the lifting platform 101 is lowered, and the support member 10 fitted with the detection container 20 is moved directly above the reader 52 by the transport device 60.

Next, as shown in FIG. 12, in the state where the support member 10 is located directly above the reader 52, the lifting platform 101 is raised to a position where detection can be performed, and positioning is performed.
The lower surface of the carrier 30 is irradiated with the excitation light 54 from the irradiator 51 through the detection container 20, and the fluorescence 55 emitted from the lower surface of the carrier 30 is input to the reader 52 through the detection container 20.
The fluorescence 55 is emitted from a fluorescent substance contained in a biologically related molecule bound to the probe on the carrier 30.
The fluorescence 55 input to the reader 52 is processed by a general method by a control device 53 (not shown in the figure) and output as image data and / or the background is removed to calculate the fluorescence intensity. Is output.

As described above, according to the method for detecting a biorelevant molecule of the biorelevant molecule, the detection kit, and the detection system of the present embodiment, the support member 10 to which the carrier 30 is attached is placed in the detection container 20 containing the anti-drying liquid 40. By fitting, the carrier 30 can be immersed in the drying prevention liquid 40 in a sealed environment.
In this state, the carrier 30 can be irradiated with the excitation light 54 via the detection container 20, and the fluorescence 55 from the carrier 30 can be input to the reader 52 via the detection container 20.
For this reason, when the detection of biologically relevant molecules is performed fully automatically by the detection system, it is possible to suppress the drying of the surface of the carrier 30 and to appropriately prevent the fluorescence detection from being inhibited (false detection). It is possible. Further, according to the configuration of the detection kit of the present embodiment, it is possible to prevent the salt from being deposited on the surface of the carrier 30, thereby enabling highly reproducible detection.

  The present invention can be suitably used when a biologically relevant molecule is automatically detected using a carrier such as a DNA chip.

DESCRIPTION OF SYMBOLS 10 Support member 11 Main body part 12 Carrier support part 13 Fitting part 20 Detection container 21 Bottom face part 22 Body part 22a Fitting part 30 Carrier 40 Drying prevention liquid 50 Detection apparatus 51 Irradiator 52 Reader 53 Control apparatus 54 Excitation light 55 Fluorescence DESCRIPTION OF SYMBOLS 60 Conveyance apparatus 61 Chuck 62 Conveyance path 70 Reaction apparatus 71 Reaction container 72 Reaction liquid 73 Heater 80 Cleaning apparatus 81 Cleaning container 82 Cleaning liquid 90 Anti-drying liquid immersion part 100 Lifting apparatus 101 Lifting stand

Claims (7)

A method for detecting a bio-related molecule that binds to a probe immobilized on a carrier,
At least a portion of the carrier on which the probe is immobilized is immersed in a detection container containing a drying prevention liquid, and excitation light is irradiated to the carrier immersed in the drying prevention liquid through the detection container. Detecting light from the biologically relevant molecule emitted by irradiation of this excitation light through the detection container. The method for detecting a bio-related molecule according to claim 1, wherein a thickness of the anti-drying liquid between the carrier and a bottom surface in the detection container is 0.1 mm to 3 mm.   A biologically relevant molecule detection comprising: a supporting member that supports a carrier on which a probe for detecting a biologically relevant molecule is immobilized; and a detection container that contains an anti-drying solution for immersing the carrier. For kit.   When the support member on which the carrier is supported and the detection container in which the anti-drying liquid is accommodated, at least a portion of the carrier on which the probe is immobilized is immersed in the anti-drying liquid. The kit for detecting a biorelevant molecule according to claim 3.   The surface for transmitting light from the biological molecule in the detection container is flat and has a thickness of 0.1 mm to 1.5 mm, for detecting biological molecules according to claim 3 or 4. kit.   The biological detection molecule detection kit according to any one of claims 3 to 5, wherein the detection container is made of at least one of a cycloolefin polymer, polystyrene, or polypropylene. A support member that supports a carrier on which a probe that binds to a biological molecule is immobilized;
A reaction container that contains a solution containing a bio-related molecule, immerses the carrier in this solution, and performs a binding reaction between the bio-related molecule and the probe;
A cleaning container that contains a cleaning liquid, and immerses the carrier in the cleaning liquid to remove biologically related molecules that have not bound to the probe; and
A detection container in which an anti-drying liquid is accommodated and when at least a portion of the carrier on which the probe is fixed is immersed in the anti-drying liquid when the carrier is fitted with a supporting member that is supported;
An excitation light irradiator for irradiating the carrier immersed in the drying prevention liquid with excitation light through the detection container;
A reader for detecting light from the biologically relevant molecule emitted by irradiation of the excitation light through the detection container;
A biologically relevant molecule detection system comprising: the support member, the reaction container, the cleaning container, the detection container, and a transport device that sequentially transports the excitation light irradiator and the reader. .