WO2014065221A1 - 複数種の目的物質を同時に検出又は定量するための分析方法及び分析キット - Google Patents
複数種の目的物質を同時に検出又は定量するための分析方法及び分析キット Download PDFInfo
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- WO2014065221A1 WO2014065221A1 PCT/JP2013/078406 JP2013078406W WO2014065221A1 WO 2014065221 A1 WO2014065221 A1 WO 2014065221A1 JP 2013078406 W JP2013078406 W JP 2013078406W WO 2014065221 A1 WO2014065221 A1 WO 2014065221A1
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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
- G01N33/54306—Solid-phase reaction mechanisms
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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
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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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
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
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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
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
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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
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N2021/7793—Sensor comprising plural indicators
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/575—Hormones
- G01N2333/59—Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
Definitions
- the present invention relates to an analysis method and an analysis kit for simultaneously detecting or quantifying a plurality of types of target substances.
- BIST Beads Array In Single Tip
- Non-patent Document 1 BIST is a cylindrical tip (hereinafter referred to as a “capillary”) with a probe bead with a diameter of 1 mm that immobilizes biomolecules such as antibodies, antigens, or DNA fragments that bind to analytes such as antigens and genes. It is a device encapsulated side by side.
- this BIST can be fitted to the nozzle of a magnetization device
- the same automation device as nucleic acid extraction for example, Magtration (registered trademark) System 6GC, Magtration (registered trademark) System 12GC Plus (manufactured by Precision System Science))
- Magtration registered trademark
- System 6GC Magtration
- System 12GC Plus manufactured by Precision System Science
- the generated signal is detected by a scanner (for example, BISTnner (manufactured by Precision System Science)).
- TSH thyroid stimulating hormone
- FT3 Free Triiodo thyronine
- FT4 Free thyroxine
- required detection density range is extensive (TSH: 0.05- 50uIu / mL
- FT3 0.5-20pg / mL
- FT4 0.1-10ng / dL, about 10 3 dynamic range).
- the present invention aims to localize signal light from probe beads in order to simultaneously detect or quantify a plurality of types of target substances.
- the gist of the present invention is as follows. (1) A plurality of carriers housed in a single container and fixed with substances for capturing a plurality of types of target substances each emit light, and the light emission is measured from the outside of the container. An analysis method for simultaneously detecting or quantifying a target substance, wherein the light emission from a desired carrier is selectively received by measuring the light emission from the carrier through an optical filter. (2) The method according to (1), wherein the optical filter is installed on the surface of the container, or the container itself is an optical filter. (3) The method according to (1) or (2), wherein the container itself is an optical filter. (4) The method according to any one of (1) to (3), wherein an optical filter having a light transmittance of 70% or less in a light emission wavelength band to be measured is used.
- the optical filter The analysis kit according to (8), wherein the container itself is an optical filter.
- the signal light from the probe beads to which the substance for capturing the target substance is fixed is localized and the signal light from other probe beads is localized. Interference with can now be suppressed.
- This specification includes the contents described in the specification and / or drawings of Japanese Patent Application No. 2012-232488, which is the basis of the priority of the present application.
- FIG. 2-1 shows the emission peak derived from the probe bead in the center of the capillary in TSH (0.5 uIU / mL) measurement by BIST.
- Fig. 2-2 shows the emission peak derived from the probe bead in the center of the capillary in TSH (25 uIU / mL) measurement by BIST.
- Figure 2-3 shows the relationship between the absorbance of the capillary material and the value of the signal light peak.
- Fig. 3-1 (upper left): A graph in which the data in Fig. 2-2 is normalized by the peak value and the vertical axis is displayed on the log scale.
- multiple carriers housed in a single container and fixed with substances for capturing multiple types of target substances each emit light, and this luminescence is measured from the outside of the container.
- an analysis method for simultaneously detecting or quantifying target substances of the above wherein the light emission from a desired carrier is selectively received by measuring the light emission from the carrier through an optical filter.
- the container may be anything as long as light emission can be measured from the outside of the container, but is preferably translucent and is preferably a material that transmits light such as resin or glass.
- the shape of the container is not particularly limited, but may be a shape such as a thin tube or pipette.
- the container is a cylindrical chip capable of sucking and discharging the liquid contained therein.
- the diffusion of signal light can be reduced by increasing the transparency of the container.
- the haze value of the container having a thickness of 1 mm is preferably 20% or less, preferably 10% or less, and more preferably 1% or less.
- Examples of the material of the container having a haze value of 20% or less with a thickness of 1 mm include polypropylene, polycarbonate, polystyrene, polymethyl methacrylate, cyclic polyolefin, polymethylpentene and the like.
- the haze value is a value representing the degree of transparency, and is defined as a ratio of diffuse transmittance to total light transmittance in JIS K7136.
- the refractive index of the container in the wavelength band of light emission or emission is preferably 1.6 or less, preferably 1.55 or less, and more preferably 1.50 or less.
- the material of the container having a refractive index of 1.6 or less include polypropylene, polycarbonate, polystyrene, polymethyl methacrylate, cyclic polyolefin, polymethylpentene, and the like.
- the target substance can be a substance to be analyzed contained in biological samples such as blood, serum, urine, lymph, mucous membranes in the mouth, nails, sputum, and samples such as river water, lake water, seawater, and soil.
- biological substances such as nucleic acids, proteins, antigens, antibodies, enzymes, sugars, bacteria such as Salmonella and Escherichia coli, metals such as cobalt, iron, molybdenum, copper, zinc, arsenic, cadmium, and vanadium. can do.
- the target substance may be labeled with a measurable label
- examples of the measurable label include a fluorescent label, a chemiluminescent label, an electrochemiluminescent label, an enzyme label, a radioactive label, and a magnetic label.
- labeling substances include Marine Blue, Cascade Blue, Cascade Yellow, Fluorescein, Rhodamine, Phycoerythrin, CyChrome, PerCP, Texas Red, Allophycocyanin, PharRed, Oregon Green-488, and Cy dyes (for example, Cy2, Cy3, Cy3.5, Cy5, Cy7 etc.), Alexa dyes (eg Alexa-488, Alexa-532, Alexa-546, Alexa-633, Alexa-680, Alexa-700, Alexa-750 etc.), BODIPY dyes (eg BODIPY FL , BODIPY TR-, etc.), luminol, acridine, adamantyldiokitacene, chemiluminescent materials such as
- the target substance may be labeled directly or indirectly.
- the sandwich ELISA method is used. Specifically, after a target substance that is an antigen is bound to a primary antibody fixed to a carrier, a biotin-labeled antibody that is a secondary antibody is bound to the target substance. Furthermore, streptavidinated HRP that catalyzes chemiluminescence binds to this secondary antibody, and labels the target substance.
- the substance for capturing the target substance may be any substance that binds to the target substance, and examples thereof include antibodies, antigens, DNA fragments, metal complexes, and other low molecular weight compounds, but are not limited thereto. is not.
- the carrier may have any shape such as irregular particles, spheres (beads), threads (strands), magnetic beads, etc., but is preferably spherical.
- a substance for capturing a different target substance may be fixed to each of the sphere (bead) or magnetic bead.
- the carrier takes the shape of a thread, it is preferable that a substance for capturing a different target substance is fixed to each of the different positions of the thread.
- the carrier takes the form of a sphere (bead).
- the sphere (bead) is preferably about 1 mm in size and is preferably made of a material such as plastic or ceramic. Such a sphere (bead) is disclosed in JP 2000-346842 A and JP 14-534657 A. It is described in gazettes.
- ⁇ Magnetic beads should have a size of several tens of ⁇ m and are made magnetic by mixing iron powder or the like into plastic or ceramic. Such magnetic beads are described in JP-A-8-62224 (Patent No. 3115501), International Publication WO96 / 29602, International Publication WO97 / 44671, and the like.
- the yarn (strand) has a diameter of about 0.1 mm and is preferably made of a resin-based material.
- Such yarn is disclosed in JP 2006-214759 A, International Publication WO01 / 53831 Pamphlet, International Publication WO2003. It is described in / 7901 pamphlet.
- the carrier may be arranged in one or two dimensions.
- the one-dimensional arrangement of the carrier means a state in which the positional information necessary for specifying the carrier from which the observed emission signal is derived is one-dimensional, for example, BIST It can be said that the carriers (beads) are arranged one-dimensionally.
- the carrier is arranged two-dimensionally means a state in which the positional information necessary for specifying the carrier from which the observed emission signal is derived is two-dimensional, for example, A measurement kit in which a carrier is spread in a single layer on a plate can be said to be a carrier that is two-dimensionally arranged.
- the substance for capturing the target substance is fixed to the carrier by covalent bond, chemical adsorption, physical adsorption, electrical interaction, hydrophobic interaction, van der Waals force, hydrogen bond and the like.
- the analysis of the target substance can be performed by bringing a substance for capturing the target substance into contact with the target substance in a state immersed in a liquid, and then measuring the emitted light.
- light emitted from a desired carrier can be selectively received by measuring light emitted from the carrier via an optical filter.
- “selectively receiving light emitted from a desired carrier” means, for example, light propagating through a capillary wall in BIST or light incident obliquely with respect to the normal of the light receiving surface.
- it also includes reducing the effect of background light.
- optical crossing of light emission signals from a plurality of carriers means a state in which light emission signals from a plurality of carriers are simultaneously received by a detector and the light emission signals from the respective carriers cannot be separated.
- Measured light emission through an optical filter can reduce signal light diffusion.
- the optical filter may have a light transmittance of 70% or less in the light emission wavelength band of the measurement target, and preferably has a light transmittance of 8% to 70%, more preferably 20% in the light emission wavelength band of the measurement target. % To 60%.
- the optical filter is made of polypropylene, polycarbonate, polystyrene, polymethyl methacrylate, cyclic polyolefin, polymethylpentene, polyethylene terephthalate and other materials such as anthraquinone colorants, perinone colorants, heterocyclic colorants, carbon black, oxidation It can be produced by coloring with a colorant such as titanium.
- the optical filter may be prepared separately from the container, or the container itself may be an optical filter. When an optical filter is prepared separately from the container, the optical filter may be installed on the surface of the container. For example, the optical filter may be installed in close contact with the container wall surface along the shape of the container wall surface.
- the luminescence can be chemiluminescence, fluorescence or electrochemiluminescence, but is chemiluminescence or fluorescence, and the emission wavelength band is preferably 340-900 nm.
- Luminescence can be measured from the outside of the container using a known technique such as a fluorimeter, spectrophotometer, scintillation counter, photodiode, CCD, CMOS, photomultiplier tube or the like.
- plural types of target substances are detected or quantified simultaneously.
- the plural kinds are at least two kinds, for example, 2, 3, 4, 5 kinds or more.
- Substances for capturing a plurality of types of target substances are respectively fixed to a plurality of carriers.
- the probe beads may include blanks and internal standards.
- a blank is a carrier on which a capture substance is not fixed.
- the internal standard is a carrier on which a substance that cannot be contained in a specimen and that captures a standard substance having a known concentration added before analysis is fixed.
- the capture substance captures the target substance specifically regardless of the presence or absence of other capture substances.
- a plurality of types of target substances can be detected simultaneously by placing the carrier in the same container and bringing a liquid specimen into contact therewith.
- the present invention emits light from a plurality of carriers, each of which is contained in a single container and to which substances for capturing a plurality of types of target substances are fixed, and measures the light emission from the outside of the container.
- An analysis kit including an optical filter for receiving light is provided. The carrier, optical filter, container, and light emission have been described above.
- BIST Bead array In Straw Tip
- TSH thyroid stimulating hormone
- the mixture was mixed by inversion several times in a sample tube with 1 ml of 1 ⁇ PBS, 0.05% Tween20 and washed. After repeating this twice, the solution was removed well, 300 ul (10 ul / bead) of 1 ⁇ PBS, 0.1% Tween20, 1% (w / w) Block Ace was added, and the mixture was allowed to stand at RT for 3 h to block.
- 2-2-1-3 After blocking, the mixture was mixed by inversion several times with 1 ml of 1 ⁇ PBS, 0.05% Tween20 and washed. After repeating this twice, the solution was removed well and filled with 1 ⁇ PBS.
- detection reagent ECL western detection reagents, GE Healthcare Japan Co., Ltd.
- PMT Photomultipulator
- Results Figures 2-1 and 2-2 show the scanning profiles (raw data) when measuring TSH with antigen concentrations of 0.5 and 25 uIU / mL by BIST, and Figure 2-3 shows the relationship between the absorbance of the capillary material and the peak value. .
- the peak value of luminescence decreased almost linearly with the increase in absorbance of the capillary material (decrease in transmittance).
- data obtained by normalizing data with an antigen concentration of 25 uIU / mL with peak values is shown in FIG. 3-1, and the relationship between the absorbance of the capillary material and the peak width is shown in FIG.
- FIG. 3-3 shows a graph plotting the relationship between the absorbance of the capillary material and the ratio of peak value to peak width (P / W ratio).
- the P / W ratio at this time was about 1.7-1.8. Further, in the region where the absorbance was higher, the P / W ratio decreased again. 4). Discussion From the results shown in Figure 3-3, coloring of the capillary material (shielding of the signal light) is an effective means for the problem of localization of the signal light (maximization of the P / W ratio) in BIST. The possibility of increasing the P / W ratio by a factor of 2 was shown.
- the method of the present invention can be used for detection and quantitative analysis of a plurality of biomolecules and low molecules that bind to biomolecules.
Abstract
Description
(1)単一の容器内に収容された、複数種の目的物質を捕獲するための物質がそれぞれ固定された複数の担体が発光し、この発光を容器外部から測定することで、複数種の目的物質を同時に検出又は定量するための分析方法であって、光学フィルターを介して担体からの発光を測定することにより、所望の担体からの発光を選択的に受光する前記方法。
(2)光学フィルターが容器表面に設置されるか、あるいは容器自体が光学フィルターとなる(1)記載の方法。
(3)容器自体が光学フィルターとなる(1)又は(2)記載の方法。
(4)測定対象の発光の波長帯における光線透過率が70%以下である光学フィルターを用いる(1)~(3)のいずれかに記載の方法。
(5)担体が球状である(1)~(4)のいずれかに記載の方法。
(6)担体が1次元的又は2次元的に配置している(1)~(5)のいずれかに記載の方法。
(7)発光が化学発光又は蛍光であり、発光の波長帯が340-900nmの範囲内にある(1)~(6)のいずれかに記載の方法。
(8)単一の容器内に収容された、複数種の目的物質を捕獲するための物質がそれぞれ固定された複数の担体が発光し、この発光を容器外部から測定することで、複数種の目的物質を同時に検出又は定量するための分析キットであって、複数種の目的物質を捕獲するための物質がそれぞれ固定された複数の担体と、所望の担体からの発光を選択的に受光するための光学フィルターとを含む前記分析キット。
(9)容器自体が光学フィルターである(8)記載の分析キット。
(10)発光が化学発光又は蛍光であり、発光の波長帯が340-900nmの範囲内にある(8)又は(9)に記載の分析キット。
本明細書は、本願の優先権の基礎である日本国特許出願、特願2012‐232488の明細書および/または図面に記載される内容を包含する。
ヘイズ値は、透明性の度合いを表す値であり、JIS K7136では、全光線透過率に対する拡散透過率の比として定義されている。
本明細書において、「所望の担体からの発光を選択的に受光する」とは、例えば、BISTにおけるキャピラリー壁を伝搬する光や、受光面の法線に対して斜めから入射する光を垂直光に比して選択的に遮光するなど、複数の担体間の光学的交差を抑制することの他、バックグラウンド光の効果を低減することも含む。
光学フィルターは、容器とは別に用意してもよいし、容器自体を光学フィルターとしてもよい。容器とは別に光学フィルターを用意する場合には、光学フィルターは容器表面に設置するとよく、例えば、容器壁面形状に沿って、容器壁面に密着させた状態で設置するとよい。
担体、光学フィルター、容器、発光については上述した。
1.要旨
全長50mm程度のキャピラリーによるTSH検出において、6濃度水準の黒色顔料により着色されたキャピラリーを用いて、TSH(抗原濃度0.5及び25uIU/mL)の検出を行った。結果、キャピラリー着色の効果は、信号光強度の高い、TSH抗原濃度25uIU/mLで顕著に現れ、ピークの縦横比((ピークの高さ)/(1/3,000幅))は、無着色時に比べ最大1.8倍程度となった。キャピラリーの着色は信号光の局在化に有効で、BISTの多項目化に対して有用な技術であることが確認された。
2.実験方法
2-1 キャピラリーの着色及び透過率の測定
住友ダウ株式会社製ポリカーボネートペレット(カリバー)を、サニーカラー株式会社製着色剤(ドライカラー Y-4-6033スモーク)によって5水準の濃度で着色(厚さ1mm、426nmに於ける透過率で約7.7%、19%、28%、62%、70%)した着色ペレットを調整した。
次いで、これらの着色ペレットを、厚さ1mmx3mm四方の板材に加工し、透過率測定用のテストピースを作製した。
このテストピースの透過率を、株式会社日立ハイテクノロジーズ社製分光光度計(U-3000)により、波長340-1000nmの領域で測定した。
2-2 TSH測定用BISTの作製
2-2-1 抗体固層ビーズの作製
2-2-1-1 プローブビーズ30個を1ml の1×PBSにてサンプルチューブ内で数回転倒混和し洗浄した。これを2回繰り返した後、液を良く取り去り、1×PBS, 20ug/ml ・TSH; TSH183を300ul(10ul/ビーズ)添加し、4℃でO/N(19h)静置し、抗体を固層した。
2-2-1-2 抗体固層後、1ml の1×PBS, 0.05% Tween20にてサンプルチューブ内で数回転倒混和し洗浄した。これを2回繰り返した後、液を良く取り去り、1×PBS, 0.1% Tween20, 1%(w/w)Block Aceを300ul(10ul/ビーズ)添加し、RTで3h静置し、ブロッキングした。
2-2-1-3 ブロッキング後、1ml の1×PBS, 0.05% Tween20にてサンプルチューブ内で数回転倒混和し洗浄した。これを2回繰り返した後、液を良く取り去り、1×PBS を満たした。
2-2-2 遮光ビーズの作製
2-2-2-1 SiCビーズを6.88g(約4000個)量り取り、15mlの1×PBSにてチューブ内で数回転倒混和し洗浄した。これを2回繰り返した後、液を良く取り去り、1×PBS, 0.1% Tween20, 1%(w/w)Block Aceを15ml添加し、RTで1hローテーターにてゆるやかに攪拌し、ブロッキングした。
2-2-2-2 ブロッキング後、15ml の1×PBS, 0.05% Tween20にてチューブ内で数回転倒混和し洗浄した。これを2回繰り返した後、液を良く取り去り、1×PBS を満たす。使用しない分は4℃保存した。
2-2-3 BISTの作製
TSHプローブビーズ1個と遮光ビーズ49個を図1のように充填した。
2-2-4 TSHの検出
2-2-4-1 下表のようにGC seriesカートリッジ(プレシジョン・システム・サイエンス株式会社製に試薬を添加し、SX-12GC(プレシジョン・システム・サイエンス株式会社製)にセットした。SX-12GCコンボラック(プレシジョン・システム・サイエンス株式会社製)のHole-2にSheath DN-100(プレシジョン・システム・サイエンス株式会社製)とともにBISTをセットし、約110分で反応が終了した。
2-2-4-2 反応が終了した後、well 1の溶液をキャピラリー内に満たし測定まで保存した。測定直前にキャピラリー内の溶液を抜き去った後、検出試薬(ECL western detection reagents, GEヘルスケアジャパン株式会社)を80ul吸引した。その後、PMT(Photomultipulator)及び走査式光ファイバー検出プローブを有するプレシジョン・システム・サイエンス株式会社製LuBEAにて測定した。
Binding Buffer: (PBS)
TSH:Human TSH(ヒト甲状腺刺激ホルモン(ヒト血清由来))
Biotin標識抗体:(抗ヒトTSH抗体をビオチン標識したもの)
Streptavidin-HRP: (Thermo scientific社製)
抗原濃度0.5及び25uIU/mLのTSHをBISTにより測定した時の走査プロファイル(生データ)を図2-1、2-2に、キャピラリー材料の吸光度とピーク値の関係を図2-3に示す。発光のピーク値はキャピラリー素材の吸光度の増加(透過率の低下)とともに、ほぼ直線的に減少した。
これに対し、抗原濃度25uIU/mLのデータをピーク値で規格化したデータを図3-1に、キャピラリー材料の吸光度とピーク幅、の関係を図3-2に示す。ピーク幅はAbs=0 から 0.5の領域で急減に減少し、それ以上の吸光度領域では、大きな変化は見られなかった。
図3-3には、キャピラリー材料の吸光度と、ピーク値とピーク幅の比(P/W比)の関係をプロットしたグラフを示す。P/W比はAbs= 0 から0.2の領域で急激に増加し、0.2から0.8の領域で極大に達し、この時のP/W比は1.7-1.8程度であった。さらに吸光度が高くなる領域では、P/W比は再度減少するという結果になった。
4.考察
図3-3の結果から、BISTにおける信号光の局在化(P/W比の最大化)という課題において、キャピラリー材料の着色(信号光の遮蔽)は有効な手段であり、適度な着色によってP/W比を2倍弱まで増大させられる可能性が示された。本結果は、信号光分布の原因の一つが、キャピラリー壁を伝搬する光であることを示しており、材料の着色によりこの信号光のキャピラリー伝搬成分を選択的に低減させると考えられる。
本明細書で引用した全ての刊行物、特許および特許出願をそのまま参考として本明細書にとり入れるものとする。
Claims (10)
- 単一の容器内に収容された、複数種の目的物質を捕獲するための物質がそれぞれ固定された複数の担体が発光し、この発光を容器外部から測定することで、複数種の目的物質を同時に検出又は定量するための分析方法であって、光学フィルターを介して担体からの発光を測定することにより、所望の担体からの発光を選択的に受光する前記方法。
- 光学フィルターが容器表面に設置されるか、あるいは容器自体が光学フィルターとなる請求項1記載の方法。
- 容器自体が光学フィルターとなる請求項1又は2記載の方法。
- 測定対象の発光の波長帯における光線透過率が70%以下である光学フィルターを用いる請求項1~3のいずれかに記載の方法。
- 担体が球状である請求項1~4のいずれかに記載の方法。
- 担体が1次元的又は2次元的に配置している請求項1~5のいずれかに記載の方法。
- 発光が化学発光又は蛍光であり、発光の波長帯が340-900nmの範囲内にある請求項1~6のいずれかに記載の方法。
- 単一の容器内に収容された、複数種の目的物質を捕獲するための物質がそれぞれ固定された複数の担体が発光し、この発光を容器外部から測定することで、複数種の目的物質を同時に検出又は定量するための分析キットであって、複数種の目的物質を捕獲するための物質がそれぞれ固定された複数の担体と、所望の担体からの発光を選択的に受光するための光学フィルターとを含む前記分析キット。
- 容器自体が光学フィルターである請求項8記載の分析キット。
- 発光が化学発光又は蛍光であり、発光の波長帯が340-900nmの範囲内にある請求項8又は9に記載の分析キット。
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US14/437,450 US10288606B2 (en) | 2012-10-22 | 2013-10-21 | Analysis method and analysis kit for simultaneously detecting or quantitating multiple types of target substances |
JP2014543273A JP6195840B2 (ja) | 2012-10-22 | 2013-10-21 | 複数種の目的物質を同時に検出又は定量するための分析方法及び分析キット |
KR1020157012755A KR20150074057A (ko) | 2012-10-22 | 2013-10-21 | 복수 종의 목적 물질을 동시에 검출 또는 정량하기 위한 분석 방법 및 분석 키트 |
CN201380054754.9A CN104737002A (zh) | 2012-10-22 | 2013-10-21 | 用于同时检测或定量多种目标物质的分析方法和分析试剂盒 |
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EP2910930A4 (en) | 2016-06-15 |
JP6195840B2 (ja) | 2017-09-13 |
US10288606B2 (en) | 2019-05-14 |
CN104737002A (zh) | 2015-06-24 |
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