WO2012124508A1 - Fluorescent light detection sheet and fluorescent light detection kit - Google Patents
Fluorescent light detection sheet and fluorescent light detection kit Download PDFInfo
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- WO2012124508A1 WO2012124508A1 PCT/JP2012/055347 JP2012055347W WO2012124508A1 WO 2012124508 A1 WO2012124508 A1 WO 2012124508A1 JP 2012055347 W JP2012055347 W JP 2012055347W WO 2012124508 A1 WO2012124508 A1 WO 2012124508A1
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- membrane
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- fluorescence detection
- autofluorescence
- detection kit
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
Definitions
- the present invention relates to a fluorescence detection sheet and a fluorescence detection kit, and more particularly to a fluorescence detection sheet and a fluorescence detection kit for detecting an analyte by fluorescence using an immunochromatography method.
- the immunochromatography method does not require heavy equipment / equipment for its determination and measurement, and is easy to operate. For example, about 5 to 10 minutes after dropping a sample solution that may contain an analyte. Measurement results can be obtained simply by standing still. Therefore, it is widely used as a simple, rapid, and highly specific determination / measurement method in many situations, for example, in clinical examinations in hospitals and in examinations in laboratories.
- the immunochromatography method is inferior to the gene amplification method (PCR method) known as a high sensitivity detection method in terms of detection sensitivity, and high sensitivity is desired.
- An immunochromatographic fluorescence detection system is an immunochromatography reagent in which an analyte and a phosphor substance are bound to a porous body (hereinafter referred to as “membrane”), and the dropped reagent is subjected to a membrane phenomenon by capillary action.
- membrane porous body
- the membrane is preferably housed in a protective case (hereinafter referred to as “housing”) made of a plastic material in order to prevent the influence of scattering when the reagent is dropped.
- housing a protective case
- a commercially available fluorescent immunochromatography kit uses a method of suppressing the autofluorescence from the housing by providing a sufficient space between the membrane and the housing by providing a column or the like. Yes.
- this method since the membrane cannot be fixed sufficiently, there is a possibility that the membrane may bend, and it is necessary to provide a sufficient space between the membrane and the housing. There are problems such as difficulty in making it easier.
- the main object of the present invention is to overcome the problems such as bending of the membrane and difficulty in miniaturization, and to suppress the generation of autofluorescence from the housing and improve the detection sensitivity.
- An object of the present invention is to provide a fluorescence detection sheet and a fluorescence detection kit for immunochromatography.
- the first aspect of the present invention It has a membrane on which a capture substance that specifically binds to the analyte is immobilized, On one side of the membrane, a light shielding sheet for blocking autofluorescence or excitation light of the housing in which the membrane is housed is attached, or a light shielding material for blocking autofluorescence or excitation light of the housing is coated.
- seat characterized by this is provided.
- a membrane on which a capture substance that specifically binds to an analyte is immobilized A housing for housing the membrane; If the surface of the membrane that receives the excitation light is the front surface and the opposite surface is the back surface, is the back surface of the membrane affixed with a light-shielding sheet that blocks autofluorescence or excitation light of the housing?
- a fluorescence detection kit is provided, which is coated with a light shielding material that blocks autofluorescence or excitation light of the housing.
- a membrane on which a capture substance that specifically binds to an analyte is immobilized A housing for housing the membrane;
- the surface of the housing that is irradiated with excitation light is affixed with a light shielding sheet that blocks autofluorescence or excitation light of the housing, or is coated with a light shielding material that blocks autofluorescence or excitation light of the housing.
- a fluorescence detection kit is provided.
- a membrane on which a capture substance that specifically binds to an analyte is immobilized A housing for housing the membrane; A light shielding sheet that blocks autofluorescence or excitation light of the housing, The fluorescent light characterized in that the light shielding sheet is disposed between the back surface of the membrane and the housing when the surface of the membrane that receives the irradiation of excitation light is the front surface and the opposite surface is the back surface.
- a detection kit is provided.
- a membrane on which a capture substance that specifically binds to an analyte is immobilized A housing for housing the membrane; There is provided a fluorescence detection kit, wherein the housing is made of a non-light emitting material that does not emit autofluorescence in a wavelength band for exciting an analysis object.
- the generation of autofluorescence from the housing can be prevented, and the detection sensitivity of the analysis object can be improved.
- the fluorescence detection kit according to a preferred embodiment of the present invention is used to detect an analyte using a known immunochromatography method, and is particularly used to detect an analyte using fluorescence emission.
- the fluorescence detection kit 1 mainly includes a fluorescence detection sheet 10 and a housing 20.
- the fluorescence detection sheet 10 includes a membrane 12, a backing sheet 16, and a light shielding sheet 14, and is housed in a housing 20 when in use.
- a capture substance (such as an antibody) that specifically binds to the analyte is fixed to the membrane 12.
- the surface of the membrane 12 is a surface that is irradiated with excitation light 50 (see FIG. 2).
- a backing sheet 16 is affixed to the membrane 12, and the membrane 12 is supported by the backing sheet 16.
- the backing sheet 16 is a sheet that improves the strength of the membrane.
- the backing sheet 16 has a smaller amount of autofluorescence than that of the housing 20.
- the backing sheet 16 is made of, for example, a polyester film.
- the backing sheet 16 may be omitted (not required).
- the light shielding sheet 14 is attached to the back surface 12 b of the membrane 12 via the backing sheet 16 and is integrated with the membrane 12.
- the light shielding sheet 14 is made of a light shielding material that blocks autofluorescence from the housing 20 and the excitation light 50 (see FIG. 2) and does not reflect the excitation light 50 (see FIG. 2).
- the light shielding sheet 14 is made of, for example, a polyester film kneaded with carbon black.
- the back surface 12b (backing sheet 16) of the membrane 12 blocks autofluorescence and excitation light 50 (see FIG. 2) from the housing 20 and reflects the excitation light 50 (see FIG. 2).
- a non-shading material may be coated (coated).
- As the light shielding material for example, a polyester resin kneaded with carbon black is used.
- the housing 20 is a plastic housing for housing the fluorescence detection sheet 10.
- the housing 20 includes an upper lid 22 and a lower container 24, both of which are integrally formed by a hinge structure.
- An observation window 26 for observing a detection line displayed on the membrane 12 (not shown) is formed at the center of the upper lid 22.
- the observation window 26 is colored and transparent, and is integrally formed of the same material as the housing 20.
- Several protrusions 28 for supporting the fluorescence detection sheet 10 from above are provided inside the upper lid 22.
- a sample addition window 30 for supplying a specimen solution 40 (see FIG. 2) containing an analysis target is provided at the end of the upper lid 22.
- a strip guard 32 for fixing the four side surfaces of the fluorescence detection sheet 10 is formed in the lower container 24 in the lower container 24, a strip guard 32 for fixing the four side surfaces of the fluorescence detection sheet 10 is formed.
- the strip guard 32 is integrally formed with the lower container 24 and is erected from the bottom surface of the lower container 24.
- the sample solution 40 is dropped onto the membrane 12 from the sample addition window 30.
- the sample solution 40 is generally composed of an analysis object (such as an antigen), a label with a fluorescent substance (such as a labeled antibody) that binds thereto, and a buffer.
- the sample solution 40 develops in the membrane 12 by capillary action, and the analyte is specifically bound to the capture substance fixed to the membrane 12 during the development. Thereafter, when a certain time has passed, the excitation light 50 is irradiated toward the membrane 12 through the observation window 26.
- the analysis target is specifically bound to the capture substance of the membrane 12
- the labeled fluorescent substance bound to the analysis target is excited to emit fluorescence, and a detection line appears (analysis). The object is detected.
- the excitation light 50 is prevented from reaching the lower container 24 of the housing 20 through the observation window 26.
- the generation of autofluorescence from the housing 20 can be suppressed, and as a result, the detection sensitivity of the analysis object can be improved.
- Conventional problems such as the occurrence of deflection and the difficulty in miniaturizing the fluorescence detection kit 1 itself can be overcome.
- the light shielding sheet 14 is separated from the membrane 12 and the backing sheet 16 and is directly attached to the bottom surface 24 a of the lower container 24.
- the bottom surface 24 a of the lower container 24 is a surface that receives the excitation light 50.
- the membrane 12 is stored in the housing 20, the membrane 12 is overlaid on the light shielding sheet 14 via the backing sheet 16.
- the material may be coated.
- the light shielding material for example, a polyester resin kneaded with carbon black is used.
- the light shielding sheet 14 is separated from the membrane 12 and the backing sheet 16 and exists alone without being attached to any member. .
- the membrane 12 is housed in the housing 20
- the light shielding sheet 14 is placed on the bottom surface 24 a of the lower container 24, and the membrane 12 is stacked on the backing sheet 16 via the backing sheet 16.
- the light shielding sheet 14 is disposed between the back surface 12 b of the membrane 12 and the housing 20.
- the housing 20 itself is made of a non-light emitting material that does not emit autofluorescence in the wavelength band that excites the analyte, and the light shielding sheet 14 Is not used.
- a non-light emitting material for example, a polyester resin kneaded with carbon black is used.
- the excitation light 50 can be prevented from reaching the lower container 24 of the housing 20 through the observation window 26, and the generation of autofluorescence from the housing 20 can be suppressed.
- the detection sensitivity of the object can be improved.
- the housing 20 has different autofluorescence depending on the type of substance. For example, if the housing 20 is made of a material kneaded with carbon, emission of autofluorescence is suppressed. However, molding the housing 20 with carbon or the like increases the material cost and processing cost, and therefore, the housing 20 is generally made of a plastic that is easy to process such as plastic. In the fluorescence detection kits 1, 3, and 5 according to the present embodiment (excluding the fluorescence detection kit 7 according to the third modification), a housing that emits autofluorescence upon receiving the excitation light 50 is used as the housing 20. Applicable to the case.
- the fluorescence detection kits 1, 3, 5, and 7 are self-contained for each member. It is assumed that the present invention is applied when the relationship between the amounts of fluorescent light satisfies the condition of formula (1). (Autofluorescence of light shielding sheet 14) ⁇ (Autofluorescence of membrane 12, backing sheet 16) ⁇ (Autofluorescence of housing 20) (1)
- the present invention can be suitably used to improve the detection sensitivity of an analysis object.
- Fluorescence detection kit 10 Fluorescence detection sheet 12 Membrane 12a Front surface 12b Back surface 14 Light shielding sheet 16 Backing sheet 20 Housing 22 Upper lid 24 Lower container 24a Bottom surface 26 Observation window 28 Projection 30 Sample addition window 32 Strip guard 40 Sample Solution 50 Excitation light
Abstract
This fluorescent light detection kit (1) has: a membrane (12) to which a capture agent that bonds specifically to an analysis subject is immobilized; a backing sheet (16) that supports the membrane (12); and a housing (20) that houses the membrane (12). When the surface of the membrane on the side that receives irradiation by excitation light is the obverse surface (12a) and the reverse side is the reverse surface (12b), a light blocking sheet (14) that blocks the excitation light or autofluorescence light of the housing (12) is applied to the reverse surface (12b) of the membrane (12).
Description
本発明は蛍光検出シートおよび蛍光検出キットに関し、特にイムノクロマト法を用いて分析対象物を蛍光により検出するための蛍光検出シートおよび蛍光検出キットに関する。
The present invention relates to a fluorescence detection sheet and a fluorescence detection kit, and more particularly to a fluorescence detection sheet and a fluorescence detection kit for detecting an analyte by fluorescence using an immunochromatography method.
イムノクロマトグラフ法は、その判定・測定に重厚な設備・機器を必要とせず、操作が簡便であり、たとえば、分析対象物を含む可能性のある検体溶液を滴下した後、約５分～１０分間静置するだけで測定結果が得られる。
そのため、簡便・迅速・特異性の高い判定・測定手法として、多くの場面で、たとえば病院における臨床検査や研究室における検定試験などで、広く使用されている。
ただ、イムノクロマトグラフ法は、検出感度の面において、高感度検出法として知られている遺伝子増幅法（ＰＣＲ法）と比べて劣っており、高感度化が望まれている。 The immunochromatography method does not require heavy equipment / equipment for its determination and measurement, and is easy to operate. For example, about 5 to 10 minutes after dropping a sample solution that may contain an analyte. Measurement results can be obtained simply by standing still.
Therefore, it is widely used as a simple, rapid, and highly specific determination / measurement method in many situations, for example, in clinical examinations in hospitals and in examinations in laboratories.
However, the immunochromatography method is inferior to the gene amplification method (PCR method) known as a high sensitivity detection method in terms of detection sensitivity, and high sensitivity is desired.
そのため、簡便・迅速・特異性の高い判定・測定手法として、多くの場面で、たとえば病院における臨床検査や研究室における検定試験などで、広く使用されている。
ただ、イムノクロマトグラフ法は、検出感度の面において、高感度検出法として知られている遺伝子増幅法（ＰＣＲ法）と比べて劣っており、高感度化が望まれている。 The immunochromatography method does not require heavy equipment / equipment for its determination and measurement, and is easy to operate. For example, about 5 to 10 minutes after dropping a sample solution that may contain an analyte. Measurement results can be obtained simply by standing still.
Therefore, it is widely used as a simple, rapid, and highly specific determination / measurement method in many situations, for example, in clinical examinations in hospitals and in examinations in laboratories.
However, the immunochromatography method is inferior to the gene amplification method (PCR method) known as a high sensitivity detection method in terms of detection sensitivity, and high sensitivity is desired.
高感度化を達成する手段として、イムノクロマト法用蛍光検出システムが提供されている（特許文献１参照）。
イムノクロマト法用蛍光検出システムとは、分析対象物と蛍光体物質とを結合させたイムノクロマト法試薬を、多孔質体（以下「メンブレン」という。）に滴下し、滴下された試薬が毛細管現象によりメンブレン内を移動し、捕捉物質固定部に固定化された分析対象物捕捉物質と接触することで前記試薬が前記捕捉物質固定部に凝縮するため、前記固定部に前記蛍光物質が蛍光発光する波長領域の励起光を照射し、前記蛍光物質が発生させた蛍光を検出部で検出することにより、被検物質の有無を定量的に判断する免疫測定法である。
特許文献１の段落００２８の記載によれば、メンブレンは、前記試薬を滴下する際に飛散の影響を防ぐために、好ましくはプラスチック材料製の保護ケース（以下「ハウジング」という。）に収納される（特許文献２参照）。 As means for achieving high sensitivity, a fluorescence detection system for immunochromatography is provided (see Patent Document 1).
An immunochromatographic fluorescence detection system is an immunochromatography reagent in which an analyte and a phosphor substance are bound to a porous body (hereinafter referred to as “membrane”), and the dropped reagent is subjected to a membrane phenomenon by capillary action. The wavelength region in which the fluorescent substance fluoresces in the fixing part because the reagent is condensed in the capturing substance fixing part by moving inside and contacting the analyte capturing substance immobilized on the capturing substance fixing part. This is an immunoassay method in which the presence or absence of a test substance is quantitatively determined by irradiating the excitation light and detecting the fluorescence generated by the fluorescent substance with a detection unit.
According to the description in paragraph 0028 of Patent Document 1, the membrane is preferably housed in a protective case (hereinafter referred to as “housing”) made of a plastic material in order to prevent the influence of scattering when the reagent is dropped. Patent Document 2).
イムノクロマト法用蛍光検出システムとは、分析対象物と蛍光体物質とを結合させたイムノクロマト法試薬を、多孔質体（以下「メンブレン」という。）に滴下し、滴下された試薬が毛細管現象によりメンブレン内を移動し、捕捉物質固定部に固定化された分析対象物捕捉物質と接触することで前記試薬が前記捕捉物質固定部に凝縮するため、前記固定部に前記蛍光物質が蛍光発光する波長領域の励起光を照射し、前記蛍光物質が発生させた蛍光を検出部で検出することにより、被検物質の有無を定量的に判断する免疫測定法である。
特許文献１の段落００２８の記載によれば、メンブレンは、前記試薬を滴下する際に飛散の影響を防ぐために、好ましくはプラスチック材料製の保護ケース（以下「ハウジング」という。）に収納される（特許文献２参照）。 As means for achieving high sensitivity, a fluorescence detection system for immunochromatography is provided (see Patent Document 1).
An immunochromatographic fluorescence detection system is an immunochromatography reagent in which an analyte and a phosphor substance are bound to a porous body (hereinafter referred to as “membrane”), and the dropped reagent is subjected to a membrane phenomenon by capillary action. The wavelength region in which the fluorescent substance fluoresces in the fixing part because the reagent is condensed in the capturing substance fixing part by moving inside and contacting the analyte capturing substance immobilized on the capturing substance fixing part. This is an immunoassay method in which the presence or absence of a test substance is quantitatively determined by irradiating the excitation light and detecting the fluorescence generated by the fluorescent substance with a detection unit.
According to the description in paragraph 0028 of Patent Document 1, the membrane is preferably housed in a protective case (hereinafter referred to as “housing”) made of a plastic material in order to prevent the influence of scattering when the reagent is dropped. Patent Document 2).
しかしながら、メンブレンをハウジングに収納するという上記のような構成では、捕捉物質固定部に励起光を照射したとき、ハウジングが自ら蛍光を発光してしまい（自家蛍光を発生してしまい）、検出部において検出対象物以外の蛍光までが検出され、その結果検出感度が低下してしまうという課題がある。
この課題に対して、既に市販化されている蛍光イムノクロマトキットでは、メンブレンとハウジングとの間に支柱などを立設して十分な間隔を設け、ハウジングからの自家蛍光を抑制する方法が用いられている。ただ、この方法によれば、メンブレンを十分に固定することができないため、メンブレンにたわみが生じてしまうおそれがあることや、メンブレンとハウジングとの間に十分な間隔を設ける必要があるため、小型化が困難になるなどの課題がある。 However, in the configuration as described above in which the membrane is housed in the housing, when the trapping substance fixing part is irradiated with excitation light, the housing itself emits fluorescence (autofluorescence is generated), and in the detection part There is a problem that even fluorescence other than the detection object is detected, and as a result, the detection sensitivity is lowered.
In response to this problem, a commercially available fluorescent immunochromatography kit uses a method of suppressing the autofluorescence from the housing by providing a sufficient space between the membrane and the housing by providing a column or the like. Yes. However, according to this method, since the membrane cannot be fixed sufficiently, there is a possibility that the membrane may bend, and it is necessary to provide a sufficient space between the membrane and the housing. There are problems such as difficulty in making it easier.
この課題に対して、既に市販化されている蛍光イムノクロマトキットでは、メンブレンとハウジングとの間に支柱などを立設して十分な間隔を設け、ハウジングからの自家蛍光を抑制する方法が用いられている。ただ、この方法によれば、メンブレンを十分に固定することができないため、メンブレンにたわみが生じてしまうおそれがあることや、メンブレンとハウジングとの間に十分な間隔を設ける必要があるため、小型化が困難になるなどの課題がある。 However, in the configuration as described above in which the membrane is housed in the housing, when the trapping substance fixing part is irradiated with excitation light, the housing itself emits fluorescence (autofluorescence is generated), and in the detection part There is a problem that even fluorescence other than the detection object is detected, and as a result, the detection sensitivity is lowered.
In response to this problem, a commercially available fluorescent immunochromatography kit uses a method of suppressing the autofluorescence from the housing by providing a sufficient space between the membrane and the housing by providing a column or the like. Yes. However, according to this method, since the membrane cannot be fixed sufficiently, there is a possibility that the membrane may bend, and it is necessary to provide a sufficient space between the membrane and the housing. There are problems such as difficulty in making it easier.
したがって、本発明の主な目的は、メンブレンにたわみが生じることや小型化が困難になるといった課題を克服したうえで、ハウジングからの自家蛍光の発生を抑制し、検出感度を向上させることができるイムノクロマト法用の蛍光検出シートおよび蛍光検出キットを提供することにある。
Therefore, the main object of the present invention is to overcome the problems such as bending of the membrane and difficulty in miniaturization, and to suppress the generation of autofluorescence from the housing and improve the detection sensitivity. An object of the present invention is to provide a fluorescence detection sheet and a fluorescence detection kit for immunochromatography.
上記課題を解決するため本発明の第１の態様によれば、
分析対象物と特異的に結合する捕捉物質が固定化されたメンブレンを有し、
前記メンブレンの片面には、前記メンブレンが収納されるハウジングの自家蛍光もしくは励起光を遮断する遮光シートが貼付されているか、または前記ハウジングの自家蛍光もしくは励起光を遮断する遮光材料がコートされていることを特徴とする蛍光検出シートが提供される。 In order to solve the above problem, according to the first aspect of the present invention,
It has a membrane on which a capture substance that specifically binds to the analyte is immobilized,
On one side of the membrane, a light shielding sheet for blocking autofluorescence or excitation light of the housing in which the membrane is housed is attached, or a light shielding material for blocking autofluorescence or excitation light of the housing is coated. The fluorescence detection sheet | seat characterized by this is provided.
分析対象物と特異的に結合する捕捉物質が固定化されたメンブレンを有し、
前記メンブレンの片面には、前記メンブレンが収納されるハウジングの自家蛍光もしくは励起光を遮断する遮光シートが貼付されているか、または前記ハウジングの自家蛍光もしくは励起光を遮断する遮光材料がコートされていることを特徴とする蛍光検出シートが提供される。 In order to solve the above problem, according to the first aspect of the present invention,
It has a membrane on which a capture substance that specifically binds to the analyte is immobilized,
On one side of the membrane, a light shielding sheet for blocking autofluorescence or excitation light of the housing in which the membrane is housed is attached, or a light shielding material for blocking autofluorescence or excitation light of the housing is coated. The fluorescence detection sheet | seat characterized by this is provided.
本発明の第２の態様によれば、
分析対象物と特異的に結合する捕捉物質が固定化されたメンブレンと、
前記メンブレンを収納するハウジングとを有し、
前記メンブレンの励起光の照射を受ける側の面を表面と、その反対面を裏面としたとき、前記メンブレンの裏面には、前記ハウジングの自家蛍光もしくは励起光を遮断する遮光シートが貼付されているか、または前記ハウジングの自家蛍光もしくは励起光を遮断する遮光材料がコートされていることを特徴とする蛍光検出キットが提供される。 According to a second aspect of the invention,
A membrane on which a capture substance that specifically binds to an analyte is immobilized;
A housing for housing the membrane;
If the surface of the membrane that receives the excitation light is the front surface and the opposite surface is the back surface, is the back surface of the membrane affixed with a light-shielding sheet that blocks autofluorescence or excitation light of the housing? Alternatively, a fluorescence detection kit is provided, which is coated with a light shielding material that blocks autofluorescence or excitation light of the housing.
分析対象物と特異的に結合する捕捉物質が固定化されたメンブレンと、
前記メンブレンを収納するハウジングとを有し、
前記メンブレンの励起光の照射を受ける側の面を表面と、その反対面を裏面としたとき、前記メンブレンの裏面には、前記ハウジングの自家蛍光もしくは励起光を遮断する遮光シートが貼付されているか、または前記ハウジングの自家蛍光もしくは励起光を遮断する遮光材料がコートされていることを特徴とする蛍光検出キットが提供される。 According to a second aspect of the invention,
A membrane on which a capture substance that specifically binds to an analyte is immobilized;
A housing for housing the membrane;
If the surface of the membrane that receives the excitation light is the front surface and the opposite surface is the back surface, is the back surface of the membrane affixed with a light-shielding sheet that blocks autofluorescence or excitation light of the housing? Alternatively, a fluorescence detection kit is provided, which is coated with a light shielding material that blocks autofluorescence or excitation light of the housing.
本発明の第３の態様によれば、
分析対象物と特異的に結合する捕捉物質が固定化されたメンブレンと、
前記メンブレンを収納するハウジングとを有し、
前記ハウジングの励起光の照射を受ける面には、前記ハウジングの自家蛍光もしくは励起光を遮断する遮光シートが貼付されているか、または前記ハウジングの自家蛍光もしくは励起光を遮断する遮光材料がコートされていることを特徴とする蛍光検出キットが提供される。 According to a third aspect of the invention,
A membrane on which a capture substance that specifically binds to an analyte is immobilized;
A housing for housing the membrane;
The surface of the housing that is irradiated with excitation light is affixed with a light shielding sheet that blocks autofluorescence or excitation light of the housing, or is coated with a light shielding material that blocks autofluorescence or excitation light of the housing. A fluorescence detection kit is provided.
分析対象物と特異的に結合する捕捉物質が固定化されたメンブレンと、
前記メンブレンを収納するハウジングとを有し、
前記ハウジングの励起光の照射を受ける面には、前記ハウジングの自家蛍光もしくは励起光を遮断する遮光シートが貼付されているか、または前記ハウジングの自家蛍光もしくは励起光を遮断する遮光材料がコートされていることを特徴とする蛍光検出キットが提供される。 According to a third aspect of the invention,
A membrane on which a capture substance that specifically binds to an analyte is immobilized;
A housing for housing the membrane;
The surface of the housing that is irradiated with excitation light is affixed with a light shielding sheet that blocks autofluorescence or excitation light of the housing, or is coated with a light shielding material that blocks autofluorescence or excitation light of the housing. A fluorescence detection kit is provided.
本発明の第４の態様によれば、
分析対象物と特異的に結合する捕捉物質が固定化されたメンブレンと、
前記メンブレンを収納するハウジングと、
前記ハウジングの自家蛍光または励起光を遮断する遮光シートとを有し、
前記メンブレンの励起光の照射を受ける側の面を表面と、その反対面を裏面としたとき、前記遮光シートが前記メンブレンの裏面と前記ハウジングとの間に配置されていることを特徴とする蛍光検出キットが提供される。 According to a fourth aspect of the invention,
A membrane on which a capture substance that specifically binds to an analyte is immobilized;
A housing for housing the membrane;
A light shielding sheet that blocks autofluorescence or excitation light of the housing,
The fluorescent light characterized in that the light shielding sheet is disposed between the back surface of the membrane and the housing when the surface of the membrane that receives the irradiation of excitation light is the front surface and the opposite surface is the back surface. A detection kit is provided.
分析対象物と特異的に結合する捕捉物質が固定化されたメンブレンと、
前記メンブレンを収納するハウジングと、
前記ハウジングの自家蛍光または励起光を遮断する遮光シートとを有し、
前記メンブレンの励起光の照射を受ける側の面を表面と、その反対面を裏面としたとき、前記遮光シートが前記メンブレンの裏面と前記ハウジングとの間に配置されていることを特徴とする蛍光検出キットが提供される。 According to a fourth aspect of the invention,
A membrane on which a capture substance that specifically binds to an analyte is immobilized;
A housing for housing the membrane;
A light shielding sheet that blocks autofluorescence or excitation light of the housing,
The fluorescent light characterized in that the light shielding sheet is disposed between the back surface of the membrane and the housing when the surface of the membrane that receives the irradiation of excitation light is the front surface and the opposite surface is the back surface. A detection kit is provided.
本発明の第５の態様によれば、
分析対象物と特異的に結合する捕捉物質が固定化されたメンブレンと、
前記メンブレンを収納するハウジングとを有し、
分析対象物を励起させる波長帯域において前記ハウジングが自家蛍光を発光しない非発光材料で構成されていることを特徴とする蛍光検出キットが提供される。 According to a fifth aspect of the present invention,
A membrane on which a capture substance that specifically binds to an analyte is immobilized;
A housing for housing the membrane;
There is provided a fluorescence detection kit, wherein the housing is made of a non-light emitting material that does not emit autofluorescence in a wavelength band for exciting an analysis object.
分析対象物と特異的に結合する捕捉物質が固定化されたメンブレンと、
前記メンブレンを収納するハウジングとを有し、
分析対象物を励起させる波長帯域において前記ハウジングが自家蛍光を発光しない非発光材料で構成されていることを特徴とする蛍光検出キットが提供される。 According to a fifth aspect of the present invention,
A membrane on which a capture substance that specifically binds to an analyte is immobilized;
A housing for housing the membrane;
There is provided a fluorescence detection kit, wherein the housing is made of a non-light emitting material that does not emit autofluorescence in a wavelength band for exciting an analysis object.
本発明によれば、ハウジングからの自家蛍光の発生が防止され、分析対象物の検出感度を向上させることができる。
According to the present invention, the generation of autofluorescence from the housing can be prevented, and the detection sensitivity of the analysis object can be improved.
以下、図面を参照しながら本発明の好ましい実施形態について説明する。
本発明の好ましい実施形態にかかる蛍光検出キットは、公知のイムノクロマト法を用いて分析対象物を検出するのに使用され、特に蛍光発光を利用して分析対象物を検出するのに使用される。 Hereinafter, preferred embodiments of the present invention will be described with reference to the drawings.
The fluorescence detection kit according to a preferred embodiment of the present invention is used to detect an analyte using a known immunochromatography method, and is particularly used to detect an analyte using fluorescence emission.
本発明の好ましい実施形態にかかる蛍光検出キットは、公知のイムノクロマト法を用いて分析対象物を検出するのに使用され、特に蛍光発光を利用して分析対象物を検出するのに使用される。 Hereinafter, preferred embodiments of the present invention will be described with reference to the drawings.
The fluorescence detection kit according to a preferred embodiment of the present invention is used to detect an analyte using a known immunochromatography method, and is particularly used to detect an analyte using fluorescence emission.
図１に示すとおり、蛍光検出キット１は主に蛍光検出シート１０とハウジング２０とから構成されている。
蛍光検出シート１０はメンブレン１２とバッキングシート１６と遮光シート１４とから構成され、使用時においてハウジング２０に収納される。
メンブレン１２には分析対象物と特異的に結合する捕捉物質（抗体など）が固定されている。メンブレン１２は表面１２ａが励起光５０（図２参照）の照射を受ける面になっている。
メンブレン１２にはバッキングシート１６が貼付されており、メンブレン１２はバッキングシート１６により支持されている。バッキングシート１６はメンブレンの強度を向上させるシートである。バッキングシート１６は自家蛍光の光量がハウジング２０のそれより小さい。バッキングシート１６はたとえばポリエステルフィルムから構成される。バッキングシート１６はなくてもよい（必須ではない。）。
遮光シート１４はメンブレン１２の裏面１２ｂに対しバッキングシート１６を介して貼付され、メンブレン１２と一体になっている。遮光シート１４はハウジング２０からの自家蛍光と励起光５０（図２参照）とを遮断し、かつ、励起光５０（図２参照）を反射しない遮光材料で構成されている。遮光シート１４は、たとえばカーボンブラックを練りこんだポリエステルフィルムから構成される。
メンブレン１２の裏面１２ｂ（バッキングシート１６）には、遮光シート１４に代えて、ハウジング２０からの自家蛍光と励起光５０（図２参照）とを遮断しかつ励起光５０（図２参照）を反射しない遮光材料がコート（被覆）されてもよい。当該遮光材料としては、たとえばカーボンブラックを練りこんだポリエステル樹脂が使用される。 As shown in FIG. 1, the fluorescence detection kit 1 mainly includes afluorescence detection sheet 10 and a housing 20.
Thefluorescence detection sheet 10 includes a membrane 12, a backing sheet 16, and a light shielding sheet 14, and is housed in a housing 20 when in use.
A capture substance (such as an antibody) that specifically binds to the analyte is fixed to themembrane 12. The surface of the membrane 12 is a surface that is irradiated with excitation light 50 (see FIG. 2).
Abacking sheet 16 is affixed to the membrane 12, and the membrane 12 is supported by the backing sheet 16. The backing sheet 16 is a sheet that improves the strength of the membrane. The backing sheet 16 has a smaller amount of autofluorescence than that of the housing 20. The backing sheet 16 is made of, for example, a polyester film. The backing sheet 16 may be omitted (not required).
Thelight shielding sheet 14 is attached to the back surface 12 b of the membrane 12 via the backing sheet 16 and is integrated with the membrane 12. The light shielding sheet 14 is made of a light shielding material that blocks autofluorescence from the housing 20 and the excitation light 50 (see FIG. 2) and does not reflect the excitation light 50 (see FIG. 2). The light shielding sheet 14 is made of, for example, a polyester film kneaded with carbon black.
Instead of thelight shielding sheet 14, the back surface 12b (backing sheet 16) of the membrane 12 blocks autofluorescence and excitation light 50 (see FIG. 2) from the housing 20 and reflects the excitation light 50 (see FIG. 2). A non-shading material may be coated (coated). As the light shielding material, for example, a polyester resin kneaded with carbon black is used.
蛍光検出シート１０はメンブレン１２とバッキングシート１６と遮光シート１４とから構成され、使用時においてハウジング２０に収納される。
メンブレン１２には分析対象物と特異的に結合する捕捉物質（抗体など）が固定されている。メンブレン１２は表面１２ａが励起光５０（図２参照）の照射を受ける面になっている。
メンブレン１２にはバッキングシート１６が貼付されており、メンブレン１２はバッキングシート１６により支持されている。バッキングシート１６はメンブレンの強度を向上させるシートである。バッキングシート１６は自家蛍光の光量がハウジング２０のそれより小さい。バッキングシート１６はたとえばポリエステルフィルムから構成される。バッキングシート１６はなくてもよい（必須ではない。）。
遮光シート１４はメンブレン１２の裏面１２ｂに対しバッキングシート１６を介して貼付され、メンブレン１２と一体になっている。遮光シート１４はハウジング２０からの自家蛍光と励起光５０（図２参照）とを遮断し、かつ、励起光５０（図２参照）を反射しない遮光材料で構成されている。遮光シート１４は、たとえばカーボンブラックを練りこんだポリエステルフィルムから構成される。
メンブレン１２の裏面１２ｂ（バッキングシート１６）には、遮光シート１４に代えて、ハウジング２０からの自家蛍光と励起光５０（図２参照）とを遮断しかつ励起光５０（図２参照）を反射しない遮光材料がコート（被覆）されてもよい。当該遮光材料としては、たとえばカーボンブラックを練りこんだポリエステル樹脂が使用される。 As shown in FIG. 1, the fluorescence detection kit 1 mainly includes a
The
A capture substance (such as an antibody) that specifically binds to the analyte is fixed to the
A
The
Instead of the
ハウジング２０は蛍光検出シート１０を収納するためのプラスチック製の筐体である。ハウジング２０は上蓋２２と下容器２４とから構成され、両者はヒンジ構造により一体成形されている。
上蓋２２の中央部には、メンブレン１２（図示略）上に表示される検出ラインを観察するための観察窓２６が形成されている。観察窓２６は着色透明であり、ハウジング２０と同一材料で一体成形されている。上蓋２２の内側には蛍光検出シート１０を上側から支持するための突起物２８が数箇所設けられている。上蓋２２の端部には、分析対象物を含む検体溶液４０（図２参照）を供給するための試料添加窓３０が設けられている。
下容器２４には、蛍光検出シート１０の４つの側面を固定するためのストリップガード３２が形成されている。ストリップガード３２は下容器２４と一体成形されており、下容器２４の底面から立設されている。 Thehousing 20 is a plastic housing for housing the fluorescence detection sheet 10. The housing 20 includes an upper lid 22 and a lower container 24, both of which are integrally formed by a hinge structure.
Anobservation window 26 for observing a detection line displayed on the membrane 12 (not shown) is formed at the center of the upper lid 22. The observation window 26 is colored and transparent, and is integrally formed of the same material as the housing 20. Several protrusions 28 for supporting the fluorescence detection sheet 10 from above are provided inside the upper lid 22. A sample addition window 30 for supplying a specimen solution 40 (see FIG. 2) containing an analysis target is provided at the end of the upper lid 22.
In thelower container 24, a strip guard 32 for fixing the four side surfaces of the fluorescence detection sheet 10 is formed. The strip guard 32 is integrally formed with the lower container 24 and is erected from the bottom surface of the lower container 24.
上蓋２２の中央部には、メンブレン１２（図示略）上に表示される検出ラインを観察するための観察窓２６が形成されている。観察窓２６は着色透明であり、ハウジング２０と同一材料で一体成形されている。上蓋２２の内側には蛍光検出シート１０を上側から支持するための突起物２８が数箇所設けられている。上蓋２２の端部には、分析対象物を含む検体溶液４０（図２参照）を供給するための試料添加窓３０が設けられている。
下容器２４には、蛍光検出シート１０の４つの側面を固定するためのストリップガード３２が形成されている。ストリップガード３２は下容器２４と一体成形されており、下容器２４の底面から立設されている。 The
An
In the
蛍光検出キット１を使用する場合は、検体溶液４０を試料添加窓３０からメンブレン１２に滴下する。検体溶液４０は通常、分析対象物（抗原など）やそれに結合する蛍光体付き標識体（標識抗体など）、緩衝液などから構成される。検体溶液４０は毛細管現象によりメンブレン１２中を展開し、その展開中において、分析対象物がメンブレン１２に固定された捕捉物質と特異的に結合される。
その後、一定時間が経過したら、観察窓２６を通じてメンブレン１２に向けて励起光５０を照射する。その結果、分析対象物がメンブレン１２の捕捉物質と特異的に結合している場合には、分析対象物と結合した標識体の蛍光体が励起され蛍光を発光し、検出ラインが出現する（分析対象物が検出される。）。 When using the fluorescence detection kit 1, thesample solution 40 is dropped onto the membrane 12 from the sample addition window 30. The sample solution 40 is generally composed of an analysis object (such as an antigen), a label with a fluorescent substance (such as a labeled antibody) that binds thereto, and a buffer. The sample solution 40 develops in the membrane 12 by capillary action, and the analyte is specifically bound to the capture substance fixed to the membrane 12 during the development.
Thereafter, when a certain time has passed, theexcitation light 50 is irradiated toward the membrane 12 through the observation window 26. As a result, when the analysis target is specifically bound to the capture substance of the membrane 12, the labeled fluorescent substance bound to the analysis target is excited to emit fluorescence, and a detection line appears (analysis). The object is detected.)
その後、一定時間が経過したら、観察窓２６を通じてメンブレン１２に向けて励起光５０を照射する。その結果、分析対象物がメンブレン１２の捕捉物質と特異的に結合している場合には、分析対象物と結合した標識体の蛍光体が励起され蛍光を発光し、検出ラインが出現する（分析対象物が検出される。）。 When using the fluorescence detection kit 1, the
Thereafter, when a certain time has passed, the
以上の実施形態によれば、蛍光検出シート１０のメンブレン１２の裏面１２ｂには遮光シート１４が貼付されているから、励起光５０が観察窓２６を通じてハウジング２０の下容器２４に到達するのが防止され、ハウジング２０からの自家蛍光の発生を抑制することができ、その結果分析対象物の検出感度を向上させることができる。
かかる構成によれば、ハウジング２０からの自家蛍光の発生を抑制するために、メンブレン１２とハウジング２０との間に支柱などを立設して十分な間隔を確保する必要もないから、メンブレン１２にたわみが生じることや蛍光検出キット１自体の小型化が困難になるといった従来の課題も克服することができる。 According to the above embodiment, since thelight shielding sheet 14 is attached to the back surface 12 b of the membrane 12 of the fluorescence detection sheet 10, the excitation light 50 is prevented from reaching the lower container 24 of the housing 20 through the observation window 26. Thus, the generation of autofluorescence from the housing 20 can be suppressed, and as a result, the detection sensitivity of the analysis object can be improved.
According to such a configuration, in order to suppress the generation of autofluorescence from thehousing 20, it is not necessary to install a column or the like between the membrane 12 and the housing 20 to ensure a sufficient interval. Conventional problems such as the occurrence of deflection and the difficulty in miniaturizing the fluorescence detection kit 1 itself can be overcome.
かかる構成によれば、ハウジング２０からの自家蛍光の発生を抑制するために、メンブレン１２とハウジング２０との間に支柱などを立設して十分な間隔を確保する必要もないから、メンブレン１２にたわみが生じることや蛍光検出キット１自体の小型化が困難になるといった従来の課題も克服することができる。 According to the above embodiment, since the
According to such a configuration, in order to suppress the generation of autofluorescence from the
［変形例１］
図３および図４に示すとおり、変形例１にかかる蛍光検出キット３では、遮光シート１４がメンブレン１２およびバッキングシート１６から分離され、下容器２４の底面２４ａに直に貼付されている。下容器２４の底面２４ａが励起光５０の照射を受ける面になっている。
メンブレン１２がハウジング２０に収納される場合には、メンブレン１２がバッキングシート１６を介して遮光シート１４の上に重ねられる。
なお、下容器２４の底面２４ａには、遮光シート１４に代えて、ハウジング２０からの自家蛍光と励起光５０（図４参照）とを遮断しかつ励起光５０（図４参照）を反射しない遮光材料がコート（被覆）されてもよい。当該遮光材料としては、たとえばカーボンブラックを練りこんだポリエステル樹脂が使用される。 [Modification 1]
As shown in FIGS. 3 and 4, in thefluorescence detection kit 3 according to the first modification, the light shielding sheet 14 is separated from the membrane 12 and the backing sheet 16 and is directly attached to the bottom surface 24 a of the lower container 24. The bottom surface 24 a of the lower container 24 is a surface that receives the excitation light 50.
When themembrane 12 is stored in the housing 20, the membrane 12 is overlaid on the light shielding sheet 14 via the backing sheet 16.
In addition, on thebottom surface 24a of the lower container 24, instead of the light-shielding sheet 14, light is blocked from blocking the self-fluorescence from the housing 20 and the excitation light 50 (see FIG. 4) and not reflecting the excitation light 50 (see FIG. 4). The material may be coated. As the light shielding material, for example, a polyester resin kneaded with carbon black is used.
図３および図４に示すとおり、変形例１にかかる蛍光検出キット３では、遮光シート１４がメンブレン１２およびバッキングシート１６から分離され、下容器２４の底面２４ａに直に貼付されている。下容器２４の底面２４ａが励起光５０の照射を受ける面になっている。
メンブレン１２がハウジング２０に収納される場合には、メンブレン１２がバッキングシート１６を介して遮光シート１４の上に重ねられる。
なお、下容器２４の底面２４ａには、遮光シート１４に代えて、ハウジング２０からの自家蛍光と励起光５０（図４参照）とを遮断しかつ励起光５０（図４参照）を反射しない遮光材料がコート（被覆）されてもよい。当該遮光材料としては、たとえばカーボンブラックを練りこんだポリエステル樹脂が使用される。 [Modification 1]
As shown in FIGS. 3 and 4, in the
When the
In addition, on the
［変形例２］
図５および図６に示すとおり、変形例２にかかる蛍光検出キット５では、遮光シート１４がメンブレン１２およびバッキングシート１６から分離され、いずれの部材にも貼付されることなく単体で存在している。
メンブレン１２がハウジング２０に収納される場合には、遮光シート１４が下容器２４の底面２４ａに載置され、その上にメンブレン１２がバッキングシート１６を介して重ねられる。遮光シート１４はメンブレン１２の裏面１２ｂとハウジング２０との間に配置される。 [Modification 2]
As shown in FIGS. 5 and 6, in thefluorescence detection kit 5 according to the modified example 2, the light shielding sheet 14 is separated from the membrane 12 and the backing sheet 16 and exists alone without being attached to any member. .
When themembrane 12 is housed in the housing 20, the light shielding sheet 14 is placed on the bottom surface 24 a of the lower container 24, and the membrane 12 is stacked on the backing sheet 16 via the backing sheet 16. The light shielding sheet 14 is disposed between the back surface 12 b of the membrane 12 and the housing 20.
図５および図６に示すとおり、変形例２にかかる蛍光検出キット５では、遮光シート１４がメンブレン１２およびバッキングシート１６から分離され、いずれの部材にも貼付されることなく単体で存在している。
メンブレン１２がハウジング２０に収納される場合には、遮光シート１４が下容器２４の底面２４ａに載置され、その上にメンブレン１２がバッキングシート１６を介して重ねられる。遮光シート１４はメンブレン１２の裏面１２ｂとハウジング２０との間に配置される。 [Modification 2]
As shown in FIGS. 5 and 6, in the
When the
［変形例３］
図７および図８に示すとおり、変形例３にかかる蛍光検出キット７では、ハウジング２０自体が分析対象物を励起させる波長帯域において自家蛍光を発光しない非発光材料から構成されており、遮光シート１４は用いられてはいない。当該非発光材料としては、たとえばカーボンブラックを練りこんだポリエステル樹脂が使用される。
メンブレン１２がハウジング２０に収納される場合には、メンブレン１２がバッキングシート１６を介して単に下容器２４の底面２４ａに載置される。 [Modification 3]
As shown in FIGS. 7 and 8, in thefluorescence detection kit 7 according to Modification 3, the housing 20 itself is made of a non-light emitting material that does not emit autofluorescence in the wavelength band that excites the analyte, and the light shielding sheet 14 Is not used. As the non-light emitting material, for example, a polyester resin kneaded with carbon black is used.
When themembrane 12 is housed in the housing 20, the membrane 12 is simply placed on the bottom surface 24 a of the lower container 24 via the backing sheet 16.
図７および図８に示すとおり、変形例３にかかる蛍光検出キット７では、ハウジング２０自体が分析対象物を励起させる波長帯域において自家蛍光を発光しない非発光材料から構成されており、遮光シート１４は用いられてはいない。当該非発光材料としては、たとえばカーボンブラックを練りこんだポリエステル樹脂が使用される。
メンブレン１２がハウジング２０に収納される場合には、メンブレン１２がバッキングシート１６を介して単に下容器２４の底面２４ａに載置される。 [Modification 3]
As shown in FIGS. 7 and 8, in the
When the
以上の変形例１～３によっても、励起光５０が観察窓２６を通じてハウジング２０の下容器２４に到達するのが防止され、ハウジング２０からの自家蛍光の発生を抑制することができ、その結果分析対象物の検出感度を向上させることができる。
According to the first to third modifications, the excitation light 50 can be prevented from reaching the lower container 24 of the housing 20 through the observation window 26, and the generation of autofluorescence from the housing 20 can be suppressed. The detection sensitivity of the object can be improved.
なお、ハウジング２０は、物質の種類によって自家蛍光の大きさが異なり、たとえばカーボンを練り込んだ素材で構成されれば自家蛍光の発光が抑制される。ただ、カーボンなどでハウジング２０を成形すると材料費、加工費が高くなるため、一般的にハウジング２０にはプラスチックなどの加工がしやすく安価なものが使用される。本実施形態にかかる蛍光検出キット１，３，５（変形例３の蛍光検出キット７は除く。）は、ハウジング２０として、励起光５０の照射を受けて自家蛍光を発光するハウジングが使用される場合に適用される。
詳しくは、ハウジング２０も含め、基本的には地球上に存在するすべての物質において自家蛍光が発光されるため、本実施形態にかかる蛍光検出キット１，３，５，７は、各部材の自家蛍光の光量の関係が式（１）の条件を満たす場合に、本願発明に適用されるものとする。
（遮光シート１４の自家蛍光）＜（メンブレン１２，バッキングシート１６の自家蛍光）＜（ハウジング２０の自家蛍光） … （１） Thehousing 20 has different autofluorescence depending on the type of substance. For example, if the housing 20 is made of a material kneaded with carbon, emission of autofluorescence is suppressed. However, molding the housing 20 with carbon or the like increases the material cost and processing cost, and therefore, the housing 20 is generally made of a plastic that is easy to process such as plastic. In the fluorescence detection kits 1, 3, and 5 according to the present embodiment (excluding the fluorescence detection kit 7 according to the third modification), a housing that emits autofluorescence upon receiving the excitation light 50 is used as the housing 20. Applicable to the case.
Specifically, since autofluorescence is emitted from all substances existing on the earth including thehousing 20, the fluorescence detection kits 1, 3, 5, and 7 according to the present embodiment are self-contained for each member. It is assumed that the present invention is applied when the relationship between the amounts of fluorescent light satisfies the condition of formula (1).
(Autofluorescence of light shielding sheet 14) <(Autofluorescence ofmembrane 12, backing sheet 16) <(Autofluorescence of housing 20) (1)
詳しくは、ハウジング２０も含め、基本的には地球上に存在するすべての物質において自家蛍光が発光されるため、本実施形態にかかる蛍光検出キット１，３，５，７は、各部材の自家蛍光の光量の関係が式（１）の条件を満たす場合に、本願発明に適用されるものとする。
（遮光シート１４の自家蛍光）＜（メンブレン１２，バッキングシート１６の自家蛍光）＜（ハウジング２０の自家蛍光） … （１） The
Specifically, since autofluorescence is emitted from all substances existing on the earth including the
(Autofluorescence of light shielding sheet 14) <(Autofluorescence of
本発明は、分析対象物の検出感度を向上させるのに好適に利用することができる。
The present invention can be suitably used to improve the detection sensitivity of an analysis object.
１，３，５，７ 蛍光検出キット
１０ 蛍光検出シート
１２ メンブレン
１２ａ 表面
１２ｂ 裏面
１４ 遮光シート
１６ バッキングシート
２０ ハウジング
２２ 上蓋
２４ 下容器
２４ａ 底面
２６ 観察窓
２８ 突起物
３０ 試料添加窓
３２ ストリップガード
４０ 検体溶液
５０ 励起光 1, 3, 5, 7Fluorescence detection kit 10 Fluorescence detection sheet 12 Membrane 12a Front surface 12b Back surface 14 Light shielding sheet 16 Backing sheet 20 Housing 22 Upper lid 24 Lower container 24a Bottom surface 26 Observation window 28 Projection 30 Sample addition window 32 Strip guard 40 Sample Solution 50 Excitation light
１０ 蛍光検出シート
１２ メンブレン
１２ａ 表面
１２ｂ 裏面
１４ 遮光シート
１６ バッキングシート
２０ ハウジング
２２ 上蓋
２４ 下容器
２４ａ 底面
２６ 観察窓
２８ 突起物
３０ 試料添加窓
３２ ストリップガード
４０ 検体溶液
５０ 励起光 1, 3, 5, 7
Claims (15)
- 分析対象物と特異的に結合する捕捉物質が固定化されたメンブレンを有し、
前記メンブレンの片面には、前記メンブレンが収納されるハウジングの自家蛍光もしくは励起光を遮断する遮光シートが貼付されているか、または前記ハウジングの自家蛍光もしくは励起光を遮断する遮光材料がコートされていることを特徴とする蛍光検出シート。 It has a membrane on which a capture substance that specifically binds to the analyte is immobilized,
On one side of the membrane, a light shielding sheet for blocking autofluorescence or excitation light of the housing in which the membrane is housed is attached, or a light shielding material for blocking autofluorescence or excitation light of the housing is coated. A fluorescence detection sheet characterized by that. - 請求項１に記載の蛍光検出シートにおいて、
前記メンブレンには、前記メンブレンを支持するためのバッキングシートが貼付されていることを特徴とする蛍光検出シート。 In the fluorescence detection sheet according to claim 1,
A fluorescence detection sheet, wherein a backing sheet for supporting the membrane is attached to the membrane. - 請求項２に記載の蛍光検出シートにおいて、
前記バッキングシートの自家蛍光が前記ハウジングの自家蛍光よりも光量が小さいことを特徴とする蛍光検出シート。 In the fluorescence detection sheet according to claim 2,
The fluorescence detection sheet, wherein the self-fluorescence of the backing sheet has a light amount smaller than the auto-fluorescence of the housing. - 分析対象物と特異的に結合する捕捉物質が固定化されたメンブレンと、
前記メンブレンを収納するハウジングとを有し、
前記メンブレンの励起光の照射を受ける側の面を表面と、その反対面を裏面としたとき、前記メンブレンの裏面には、前記ハウジングの自家蛍光もしくは励起光を遮断する遮光シートが貼付されているか、または前記ハウジングの自家蛍光もしくは励起光を遮断する遮光材料がコートされていることを特徴とする蛍光検出キット。 A membrane on which a capture substance that specifically binds to an analyte is immobilized;
A housing for housing the membrane;
If the surface of the membrane that is irradiated with excitation light is the front surface and the opposite surface is the back surface, is the back surface of the membrane affixed with a light-shielding sheet that blocks autofluorescence or excitation light of the housing? Or a fluorescence detection kit, wherein the housing is coated with a light shielding material that blocks autofluorescence or excitation light of the housing. - 請求項４に記載の蛍光検出キットにおいて、
前記メンブレンには、前記メンブレンを支持するためのバッキングシートが貼付されていることを特徴とする蛍光検出キット。 The fluorescence detection kit according to claim 4,
A fluorescence detection kit, wherein a backing sheet for supporting the membrane is attached to the membrane. - 請求項５に記載の蛍光検出キットにおいて、
前記バッキングシートの自家蛍光が前記ハウジングの自家蛍光よりも光量が小さいことを特徴とする蛍光検出キット。 The fluorescence detection kit according to claim 5,
A fluorescence detection kit, wherein the autofluorescence of the backing sheet has a smaller amount of light than the autofluorescence of the housing. - 分析対象物と特異的に結合する捕捉物質が固定化されたメンブレンと、
前記メンブレンを収納するハウジングとを有し、
前記ハウジングの励起光の照射を受ける面には、前記ハウジングの自家蛍光もしくは励起光を遮断する遮光シートが貼付されているか、または前記ハウジングの自家蛍光もしくは励起光を遮断する遮光材料がコートされていることを特徴とする蛍光検出キット。 A membrane on which a capture substance that specifically binds to an analyte is immobilized;
A housing for housing the membrane;
The surface of the housing that is irradiated with excitation light is affixed with a light shielding sheet that blocks autofluorescence or excitation light of the housing, or is coated with a light shielding material that blocks autofluorescence or excitation light of the housing. A fluorescence detection kit. - 請求項７に記載の蛍光検出キットにおいて、
前記メンブレンには、前記メンブレンを支持するためのバッキングシートが貼付されていることを特徴とする蛍光検出キット。 The fluorescence detection kit according to claim 7,
A fluorescence detection kit, wherein a backing sheet for supporting the membrane is attached to the membrane. - 請求項８に記載の蛍光検出キットにおいて、
前記バッキングシートの自家蛍光が前記ハウジングの自家蛍光よりも光量が小さいことを特徴とする蛍光検出キット。 The fluorescence detection kit according to claim 8,
A fluorescence detection kit, wherein the autofluorescence of the backing sheet has a smaller amount of light than the autofluorescence of the housing. - 分析対象物と特異的に結合する捕捉物質が固定化されたメンブレンと、
前記メンブレンを収納するハウジングと、
前記ハウジングの自家蛍光または励起光を遮断する遮光シートとを有し、
前記メンブレンの励起光の照射を受ける側の面を表面と、その反対面を裏面としたとき、前記遮光シートが前記メンブレンの裏面と前記ハウジングとの間に配置されていることを特徴とする蛍光検出キット。 A membrane on which a capture substance that specifically binds to an analyte is immobilized;
A housing for housing the membrane;
A light shielding sheet that blocks autofluorescence or excitation light of the housing,
The fluorescent light characterized in that the light shielding sheet is disposed between the back surface of the membrane and the housing when the surface of the membrane that receives the irradiation of excitation light is the front surface and the opposite surface is the back surface. Detection kit. - 請求項１０に記載の蛍光検出キットにおいて、
前記メンブレンには、前記メンブレンを支持するためのバッキングシートが貼付されていることを特徴とする蛍光検出キット。 The fluorescence detection kit according to claim 10,
A fluorescence detection kit, wherein a backing sheet for supporting the membrane is attached to the membrane. - 請求項１１に記載の蛍光検出キットにおいて、
前記バッキングシートの自家蛍光が前記ハウジングの自家蛍光よりも光量が小さいことを特徴とする蛍光検出キット。 The fluorescence detection kit according to claim 11,
A fluorescence detection kit, wherein the autofluorescence of the backing sheet has a smaller amount of light than the autofluorescence of the housing. - 分析対象物と特異的に結合する捕捉物質が固定化されたメンブレンと、
前記メンブレンを収納するハウジングとを有し、
分析対象物を励起させる波長帯域において前記ハウジングが自家蛍光を発光しない非発光材料で構成されていることを特徴とする蛍光検出キット。 A membrane on which a capture substance that specifically binds to an analyte is immobilized;
A housing for housing the membrane;
A fluorescence detection kit, wherein the housing is made of a non-light emitting material that does not emit autofluorescence in a wavelength band for exciting an analysis object. - 請求項１３に記載の蛍光検出キットにおいて、
前記メンブレンには、前記メンブレンを支持するためのバッキングシートが貼付されていることを特徴とする蛍光検出キット。 The fluorescence detection kit according to claim 13,
A fluorescence detection kit, wherein a backing sheet for supporting the membrane is attached to the membrane. - 請求項１４に記載の蛍光検出キットにおいて、
前記バッキングシートの自家蛍光が前記ハウジングの自家蛍光よりも光量が小さいことを特徴とする蛍光検出キット。 The fluorescence detection kit according to claim 14,
A fluorescence detection kit, wherein the autofluorescence of the backing sheet has a smaller amount of light than the autofluorescence of the housing.
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| WO2020013611A1 (en) * | 2018-07-11 | 2020-01-16 | 주식회사 수젠텍 | Fluorescent standard strip |
| WO2022163124A1 (en) * | 2021-01-29 | 2022-08-04 | 東洋濾紙株式会社 | Membrane for immunochromatographic assays, test strip for immunochromatographic assays, and test method |
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| CN111982869B (en) * | 2020-08-06 | 2023-09-19 | 中国科学院合肥物质科学研究院 | Calibration method and calibration accessory for quantitative fluorescence measurement |
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