Immunochromatography detection method

A technology of immunochromatography detection and chromatography media, which is applied in the direction of measuring devices, analytical materials, instruments, etc., can solve the problems of decreased detection sensitivity, abnormal function, and inability to fully suppress non-specific reactions

Inactive Publication Date: 2014-12-10
TANAKA PRECIOUS METAL IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] As protective stabilizing solutions, hitherto, those containing proteins such as bovine serum albumin (BSA) as a protective stabilizer are known, but there is a problem that they cannot be kept stable for a long period of time in a dry state, so people have Further improvements and research were carried out
[0010] On the other hand, with regard to immunochromatographic devices, there are common problems of background coloration (coloration of parts other than the stationary phase antibody at the determination part), blank coloration (coloration of the stationary phase in the absence of the detection substance) ) and prozone phenomenon (false-negative phenomenon in which when an excess of the test substance is used as a sample, it is clearly observed that the test substance seems to be present in a small amount), these problems not only reduce the SN rate at the time of detection, but also cause functional abnormalities
[0015] However, even if the non-specific reaction is suppressed, there is still a problem of a decrease in detection sensitivity, so this goal has not been satisfactorily achieved, so there is still a similar problem of not being able to sufficiently suppress the non-specific reaction

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0148] (1) Preparation of the determination part on the chromatography medium

[0149] As the membrane, a sheet made of nitrocellulose (manufactured by Millipore, trade name: HF 12250 mm×25 mm) was used. Anti-influenza A virus monoclonal antibody (primary antibody) was diluted to a concentration of 1.0 mg / ml with 10 mM phosphate buffer (pH 7.4) containing 5 mass % sucrose and 5 mass % isopropanol. The diluted solution (150 μL) was coated on a membrane with a width of 1 mm by means of an antibody coater (manufactured by BioDot Corporation), and then dried at 50° C. for 30 minutes and then at room temperature overnight to prepare a judgment portion on a chromatography medium.

[0150] (2) Preparation of marker solution

[0151] Anti-influenza A monoclonal antibody (0.1 mL) diluted to a concentration of 0.1 mg / mL with HEPES buffer (pH 7.5) was added to 0.5 mL of gold colloidal suspension (manufactured by Tanaka Kikinzoku Kogyo, average particle diameter: 40 nm) , and then let s...

Embodiment 2

[0161] The measurement was performed in the same manner as in Example 1, except that 0.01% by mass of PEG-SH (molecular weight: 2,000) was used as a protecting agent for the marker in the preparation of the marker solution in step (2) of Example 1. Judgment was made according to the same visual judgment criteria. The judgment results are shown in Table 1.

Embodiment 3

[0163] The measurement was performed in the same manner as in Example 1, except that 0.01% by mass of PEG-SH (molecular weight: 20,000) was used as a protective agent for the marker in the preparation of the marker solution in step (2) of Example 1. Judgment was made according to the same visual judgment criteria. The judgment results are shown in Table 1.

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Abstract

The present invention provides an immunochromatography detection method whereby non-specific reactions can be minimized. The present invention relates to an immunochromatography detection method including: a step of adding an analyte diluted solution containing an analyte to a chromatography medium; a step of recognizing the detection target by a labeling substance which has been modified with gold nanoparticles, and which is retained in a dried state by a labeling substance retaining part; a step of developing a complex of the labeling substance and the detection target as the mobile phase; and a step of detecting the detection target in the developed mobile phase, by an assessment part, wherein the immunochromatography detection method is characterized in that the labeling substance is protected by polyalkylene glycol having one or more mercapto groups and / or a derivative thereof, and then retained in a dried state, together with arginine and casein, in the labeling substance retaining part.

Description

technical field [0001] The invention relates to a high-performance and high-sensitivity immunochromatographic detection method for suppressing non-specific reactions. In addition, the present invention relates to a detection kit capable of rapidly, simply and accurately detecting / measuring a detection target in a sample by suppressing a non-specific reaction. [0002] More specifically, the present invention relates to a rapid, simple and accurate assay for detection in samples by protecting and stabilizing markers labeled with metal nanoparticles against drying treatments or dry states and inhibiting non-specific reactions. The method for the object, wherein the marker is used to detect the detection object (substance to be detected), that is, the marker is a sensitized metal colloid, wherein the metal colloid is bound by a substance that specifically binds to the detection object Sensitized. Background technique [0003] In recent years, the importance of dipstick-type i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
CPCG01N33/558G01N33/538G01N33/56983G01N33/54387G01N33/587G01N33/54388
Inventor 伊藤大辅
Owner TANAKA PRECIOUS METAL IND
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