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WO2012124508A1 - Feuille de détection fluorimétrique, et équipement pour détection fluorimétrique - Google Patents

Feuille de détection fluorimétrique, et équipement pour détection fluorimétrique Download PDF

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Publication number
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
Authority
WO
WIPO (PCT)
Prior art keywords
membrane
housing
fluorescence detection
autofluorescence
detection kit
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/JP2012/055347
Other languages
English (en)
Japanese (ja)
Inventor
大輔 濱中
史生 長井
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Konica Minolta Advanced Layers Inc
Original Assignee
Konica Minolta Advanced Layers Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Konica Minolta Advanced Layers Inc filed Critical Konica Minolta Advanced Layers Inc
Priority to JP2013504651A priority Critical patent/JPWO2012124508A1/ja
Publication of WO2012124508A1 publication Critical patent/WO2012124508A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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
    • 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

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  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Pathology (AREA)
  • Urology & Nephrology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Hematology (AREA)
  • Cell Biology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

L'équipement pour détection fluorimétrique (1) de l'invention possède : une membrane (12) sur laquelle est fixée une substance piégeante qui se lie spécifiquement à l'objet de l'analyse; une feuille de support (16) qui maintient la membrane (12); et un logement (20) qui admet la membrane (12). Lorsque la face côté exposé à une lumière d'excitation de la membrane (12), consiste en une face endroit (12a), et que la face opposée à celle-ci consiste en une face envers (12b), une feuille de blocage (14) qui bloque la fluorescence de la membrane (12) elle-même ou la lumière d'excitation, est collée sur la face envers (12b) de la membrane (12).
PCT/JP2012/055347 2011-03-16 2012-03-02 Feuille de détection fluorimétrique, et équipement pour détection fluorimétrique Ceased WO2012124508A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2013504651A JPWO2012124508A1 (ja) 2011-03-16 2012-03-02 蛍光検出シートおよび蛍光検出キット

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2011-057346 2011-03-16
JP2011057346 2011-03-16

Publications (1)

Publication Number Publication Date
WO2012124508A1 true WO2012124508A1 (fr) 2012-09-20

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JP (1) JPWO2012124508A1 (fr)
WO (1) WO2012124508A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2019152584A (ja) * 2018-03-05 2019-09-12 株式会社ニチレイバイオサイエンス ハウジング及び検査キット
WO2020013611A1 (fr) * 2018-07-11 2020-01-16 주식회사 수젠텍 Bande standard fluorescente
JPWO2022163124A1 (fr) * 2021-01-29 2022-08-04
CN114910453A (zh) * 2021-02-10 2022-08-16 天津九安医疗电子股份有限公司 一种一次性荧光免疫层析检测试剂盒

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111982869B (zh) * 2020-08-06 2023-09-19 中国科学院合肥物质科学研究院 一种用于定量荧光测量的校准方法及校准配件

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007003363A (ja) * 2005-06-24 2007-01-11 Matsushita Electric Ind Co Ltd プローブ担体および蛍光読み取り装置
WO2008090922A1 (fr) * 2007-01-24 2008-07-31 Toray Industries, Inc. Puce d'analyse et procédé d'analyse
WO2009099269A1 (fr) * 2008-10-24 2009-08-13 Boditechmed Inc. Système de mesure quantitative de la glycohémoglobine et procédé de mesure de la teneur en glycohémoglobine l'utilisant

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007003363A (ja) * 2005-06-24 2007-01-11 Matsushita Electric Ind Co Ltd プローブ担体および蛍光読み取り装置
WO2008090922A1 (fr) * 2007-01-24 2008-07-31 Toray Industries, Inc. Puce d'analyse et procédé d'analyse
WO2009099269A1 (fr) * 2008-10-24 2009-08-13 Boditechmed Inc. Système de mesure quantitative de la glycohémoglobine et procédé de mesure de la teneur en glycohémoglobine l'utilisant

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2019152584A (ja) * 2018-03-05 2019-09-12 株式会社ニチレイバイオサイエンス ハウジング及び検査キット
JP7133322B2 (ja) 2018-03-05 2022-09-08 株式会社ニチレイバイオサイエンス ハウジング及び検査キット
WO2020013611A1 (fr) * 2018-07-11 2020-01-16 주식회사 수젠텍 Bande standard fluorescente
KR20200006755A (ko) * 2018-07-11 2020-01-21 주식회사 수젠텍 형광 표준 스트립
KR102074471B1 (ko) 2018-07-11 2020-02-06 주식회사 수젠텍 형광 표준 스트립
US11442011B2 (en) 2018-07-11 2022-09-13 Sugentech, Inc. Fluorescent standard strip
JPWO2022163124A1 (fr) * 2021-01-29 2022-08-04
WO2022163124A1 (fr) * 2021-01-29 2022-08-04 東洋濾紙株式会社 Membrane pour dosages immunochromatographiques, bandelette réactive pour dosages immunochromatographiques et procédé de test
JP7683852B2 (ja) 2021-01-29 2025-05-27 東洋濾紙株式会社 イムノクロマトアッセイ用メンブレン、イムノクロマトアッセイ用テストストリップ、および検査方法
CN114910453A (zh) * 2021-02-10 2022-08-16 天津九安医疗电子股份有限公司 一种一次性荧光免疫层析检测试剂盒

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