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Immunoassay method and immunoassay kit to be used therein

An immunoassay, kit technology, applied in immunoglobulins, chemical instruments and methods, specific peptides, etc., which can solve the problems of false positives and inability to obtain sufficient effects.

Inactive Publication Date: 2008-03-19
ARKRAY INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, when analyzing samples (such as throat swabs, etc.) using immunoassay methods such as latex agglutination, there is a problem of false positives
That is, there is a problem that although the substance to be measured does not exist in the sample, the particles are bound and aggregated due to a non-specific reaction, so it is judged as positive
However, this method exhibits its effect in EIA (enzyme immunoassay) using a membrane solid phase, but cannot obtain sufficient effect in an immunoassay method using antibody-sensitized particles such as the latex agglutination method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1. Preparation of Anti-hemolytic Streptococcus Antibody Sensitization Latex Reagent

[0039] Using anti-hemolytic streptococcus rabbit polyclonal antibody and 50 mM boric acid buffer (pH 7.4), prepare antibody solution (antibody concentration: 0.7 mg / mL). 0.5 mL of the above antibody solution was mixed with 0.5 mL of 1% (w / v) polystyrene latex particles (manufactured by Sekisui Chemical Co., Ltd., with an average particle size of 0.5 μm), and incubated at 37° C. for 1 hour to render the above anti-hemolytic Sexual Streptococcus rabbit polyclonal antibody was sensitized to the above polystyrene latex particles. Next, BSA (manufactured by Sigma Corporation) was added so as to be 1% by weight. Incubate for 1 hour at 37°C for blocking. After washing the blocked antibody-sensitized latex particles with a phosphate buffer, the latex particles were dispersed in a latex dispersion buffer (50 mM Tris-HCl buffer (pH 8.4), 0.1% by weight BSA, 0.1% by weight NaN 3 ), Anti-hemoly...

Embodiment 2

[0065] 1. Preparation of Anti-Influenza Type A Antibody Sensitization Latex Reagent

[0066] Antibody solution (antibody concentration: 0.8 mg / mL) was prepared using anti-influenza type A mouse monoclonal antibody and 50 mM phosphate buffer (pH: 7.4). In the same manner as the anti-hemolytic streptococcus antibody-sensitized latex reagent of Example 1 above, the anti-influenza type A antibody-sensitized latex reagent was prepared using the above-mentioned antibody solution.

[0067] 2. Determination of influenza A antigen by latex agglutination method

[0068] As the antigen, purified influenza A virus A / Kiev / 301 / 94 (H3N2) was used. The above-mentioned antigen was diluted with the sample diluent used in Example 1, and two kinds of antigen concentrations (4 μg / mL and 20 μg / mL) were prepared. Next, immediately before the measurement, the above-mentioned antigen solution of each concentration was diluted 10-fold with a sample extract solution having the following composition, a...

Embodiment 2-1

[0089] (Example 2-1: Condition 1)

[0090] The above suspension was added to COSMOGEL (registered trademark) 30SP (trade name) (cation exchange carrier (functional group: sulfonic acid group, particle size: 10 μm); manufactured by NACALAI TESQUE) and left for 2 minutes to perform cation exchange treatment. Next, the change in absorbance at a measurement wavelength of 660 nm was measured in the same manner as in the above "2. Measurement of hemolytic streptococcal antigen by latex agglutination method".

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Abstract

In an immunoassay method with the use of antibody-sensitized particles, it is intended to prevent the occurrence of a false positive reaction. An immunoassay method which comprises mixing a specimen with antibody-sensitized particles having been sensitized with an antibody capable of reacting a subject to be assayed in the specimen, measuring the aggregation of the antibody-sensitized particles due to the antigen-antibody reaction and thus detecting or quantifying the subject to be assayed in the specimen, wherein the specimen is pretreated with a cation exchanger before the mixing. As the cation exchanger as described above, use can be made of, for example, a cation exchanger having sulfonate or carboxyl group as a functional group. It is not restricted in form. Namely, it may be a cation exchange filter, a cation exchange column or the like.

Description

technical field [0001] The present invention relates to an immunoassay method and an immunoassay kit used in the method. Background technique [0002] In recent years, simple test reagents and kits have been developed that use antigen-antibody reactions to detect or quantify items within a few minutes to tens of minutes from the start of the measurement. When the measurement item is an infectious disease, if the presence or absence of infection in a sample collected from a patient can be judged on the spot, the patient can be treated at an early stage of symptoms. Therefore, the importance of such simple inspection reagents and kits is increasing year by year in the medical field. [0003] Currently, immunoassay methods such as latex agglutination are widely used as one of simple inspection methods. Latex agglutination method is a method of using antigen-antibody reaction to cause antibody-sensitized latex particles to bind and agglutinate with the target antigen in the sp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/569
CPCG01N33/5306G01N33/54313C07K16/1018C07K16/1275G01N33/54393
Inventor 大宫一纮大代京一
Owner ARKRAY INC
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