Method for measuring concentration of test substance, and detection apparatus

Patent metadata

Field	Value
Publication number	US-2016305889-A1
Application number	US-201415030183-A
Country	US
Kind code	A1
Filing date	Oct 20, 2014
Priority date	Oct 21, 2013
Publication date	Oct 20, 2016
Grant date	—

Abstract

The present invention provide a method for detecting a test substance utilizing a color development caused by an enzyme reaction, or a color development caused by a specific interaction such as an antigen-antibody reaction and an enzyme reaction, the test substance being detected rapidly, sensitively and quantitatively without using a spectroscopic measurement device. The method for measuring a concentration of a test substance by the present invention comprises the steps of: generating a peroxide from the test substance; obtaining a polymerized substance by bringing an oxidoreductase for producing a polymerized substance and a substrate of the oxidoreductase for producing a polymerized substance into contact with the peroxide; and irradiating the polymerized substance with light to record a temporal variation information of an intensity of scattered light generated from an irradiation point.

First claim

1 - 13 . (canceled)

14 . A method for measuring a concentration of a test substance, the method comprising the steps of:
obtaining a polymerized substance by bringing a modifier in which a substance having a specific interaction with the test substance is modified with an oxidoreductase for producing a polymerized substance into contact with the test substance, and then bringing a peroxide and a substrate of the oxidoreductase for producing a polymerized substance of the substrate into contact with the test substance; and irradiating the polymerized substance with light to record a temporal variation information of an intensity of scattered light generated from an irradiation point.

15 . The method for measuring a concentration of a test substance according to claim 14 , wherein the specific interaction with the test substance is an antigen-antibody reaction.

16 . The method for measuring a concentration of a test substance according to claim 14 , wherein the temporal variation information constitutes a signal waveform, the method further comprising the step of identifying a time taken from a predetermined time point on and after a start of irradiation of the light to the test substance until the signal waveform shows an extremum.

17 . The method for measuring a concentration of a test substance according to claim 14 , wherein the obtaining step of a polymerized substance is performed on a base.

18 . The method for measuring a concentration of a test substance according to claim 17 ,
wherein a first base on which at least one of group X substances consisting of the test substance and the substance having a specific interaction with the test substance exists, and a second base on which at least one of group X substances consisting of the test substance and the substance having a specific interaction with the test substance does not exist are stacked, and irradiation of light is performed from the second base side.

19 . The method for measuring a concentration of a test substance according to claim 17 ,
wherein the base comprises a group X substance existing region where at least one of group X substances consisting of the test substance and the substance having a specific interaction with the test substance exists, and a group X substance non-existing region where the group X substance does not exist, and the group X substance non-existing region is irradiated with the light.

20 . The method for measuring a concentration of a test substance according to claim 19 ,
wherein a porous support is provided on the base, and the group X substance is immobilized with the porous support.

21 . A detection apparatus for detecting a test substance comprising:
a light source allowing light to enter the test substance; a photoelectric conversion element detecting scattered light generated from a polymerized substance derived from the test substance; and a recording medium continuously recording a signal output from the photoelectric conversion element during a predetermined period of time, wherein the polymerized substance is obtained by bringing a modifier in which a substance having a specific interaction with the test substance is modified with an oxidoreductase for producing a polymerized substance into contact with the test substance, and then bringing a peroxide and a substrate of the oxidoreductase for producing a polymerized substance of the substrate into contact with the test substance.

22 . The detection apparatus according to claim 21 ,
wherein the polymerized substance derived from the test substance exists on a first surface side of a light-transmitting base, and wherein the detection apparatus further comprises a lens facing a second surface side of the light transmitting base.

23 . The detection apparatus according to claim 21 further comprising a calculating means identifying a time taken from a predetermined time point on and after a start of irradiation of the light to the polymerized substance derived from the test substance until a signal waveform recorded in the recording medium shows an extremum.

24 . The detection apparatus according to claim 21 comprising, on the first surface side of the light transmitting base, a group X substance existing region where at least one of group X substances consisting of the polymerized substance derived from the test substance and a substance having a specific interaction with the polymerized substance derived from the test substance exists, and a group X substance non-existing region where the group X substance does not exist.

25 . The method for measuring a concentration of a test substance according to claim 14 ,
wherein the recording step comprising steps of obtaining a polymer that scatters light by irradiating the polymerizable substance with light and allowing the polymerizable substance to be aggregated; recording a temporal variation information of an intensity of scattered light generated from an irradiation in the polymer.

26 . The method for measuring a concentration of a test substance according to claim 14 ,
wherein the recording step of a temporal variation information comprising recording a temporal variation information of an intensity of scattered light generated from an irradiation, based on the phase difference of two lights beginning at the polymer.

27 . The method for measuring a concentration of a test substance according to claim 14 , comprising further steps of
wherein the temporal variation information constitutes a signal waveform, the method further comprising the step of identifying a time taken from a predetermined time point on and after a start of irradiation of the light to the test substance until the signal waveform shows an extremum.

28 . The method for measuring a concentration of a test substance according to claim 14 ,
wherein the obtaining step of a polymerized substance is performed on a base.

Assignees

• Univ Osaka

Inventors

• Yoshikawa Hiroyuki
• Tamiya Eiichi
• Imura Shuhei

Classifications

• G01N2201/062

  LED's · CPC title

• C12Q1/26

  involving oxidoreductase · CPC title

• G01N21/78

  producing a change of colour · CPC title

• G01N2333/904

  acting on CHOH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1) · CPC title

• G01N21/82 Primary

  producing a precipitate or turbidity · CPC title