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Insoluble carrier particle nephelometric immunoassay reagent

A carrier particle and immunoassay technology, applied in biological testing, measuring devices, instruments, etc., can solve problems such as unstable sensitivity and surface charge changes

Inactive Publication Date: 2006-07-12
KYOWA MEDEX CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] As mentioned above, latex is often used in the measurement system of immunoagglutination reaction such as latex turbidimetric immunoassay, and the latex suspension refers to the reaction of plasma components coexisting with the reaction solution by mixing a trace amount of heavy metal ions etc. into it. , to change the adsorption of the latex carrier of the antigen or antibody, or to dissociate the antigen or antibody from the latex bound to the antigen or antibody to generate free antigen or antibody. During the reaction, the degree of agglutination of the latex occurs Changes, there is a problem of unstable sensitivity
In addition, for latex that is not bound to antigen or antibody, carboxyl or sulfonic acid groups with surface charge will change the surface charge and unstable sensitivity due to the binding of heavy metal ions.

Method used

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  • Insoluble carrier particle nephelometric immunoassay reagent

Examples

Experimental program
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Effect test

Embodiment 1

[0044] [Preparation of reagent 1 (buffer solution)]

[0045] As a buffer, 3.26 g of N-bis(hydroxyethyl)glycine (manufactured by Dojin Chemical Co., Ltd.) was dissolved in distilled water, and 0.1 g of surfactant (Triton) X-100, 17.5 g of sodium chloride, and 0.01 g of sodium chloride were added respectively. When measuring the pH at 20°C, add 1 mol / L hydrochloric acid or sodium hydroxide aqueous solution to adjust the pH to 8.0, and add distilled water to make the total volume 1,000ml. Also available are 3.58 g of tricoflavone (manufactured by Tongren Chemical Co.), TAPSO {2-hydroxy-3-[N-tris(hydroxymethyl)methylamino]propanesulfonic acid} (manufactured by Tongren Chemical Co.) 5.18 g, POPSO [piperazine -1,4-bis(2-hydroxy-3-propanesulfonic acid) dihydrate] (manufactured by Dojin Chemical Co., Ltd.) 7.97 g, TES{2-[N-tris(hydroxymethyl)methylamino]propanesulfonic acid} (Manufactured by Dojin Chemical Co., Ltd.) 4.59 g instead of N-bis(hydroxyethyl)glycine was dissolved in disti...

Embodiment 2

[0057] [Preparation of reagent 3 (latex suspension)]

[0058] Dissolve 3.26 g of N-bis(hydroxyethyl)glycine as a buffer in distilled water, add 3.3 ml of 10% latex (particle size: 0.087 μm; manufactured by Sekisui Chemical Co., Ltd.) and 0.1 g of sodium azide, at 20° C. While measuring the pH, add 1 mol / L hydrochloric acid or sodium hydroxide aqueous solution to adjust the pH to 7.8, and add distilled water to make the total volume 1,000ml. Instead of N-bis(hydroxyethyl)glycine as a buffer, 3.58g of tricoflavone, 5.18g of TAPSO, 7.97g of POPSO, and 4.59g of TES were dissolved in distilled water respectively, and latex and sodium lamination were also added to adjust the pH to 7. After 8, add distilled water to a full volume of 1,000ml. In this way, prepare the 20mmol / L buffer solution containing N-bis(hydroxyethyl)glycine or tricoflavone in the present invention, and the comparative example is the 20mmol / L buffer solution containing TAPSO, POPSPO, TES respectively, and prepare...

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Abstract

The present invention provides: a reagent and a kit for an insoluble carrier particle nephelometric immunoassay which stabilize the agglutination reaction by suppressing the action of blood plasma components that are involved in the agglutination reaction of insoluble carrier particles such as latex and affect the values to be determined, to provide the stable absorbances of the reaction solutions and the accurate determination results; and a method of an insoluble carrier particle nephelometric immunoassay utilizing said reagent or kit. An antigen or an antibody is carried on an insoluble carrier particle in the presence or absence of a buffer solution comprising a compound having within its molecule a group shown below such as bicine, tricine, then an antibody- or antigen-sensitized insoluble carrier particle suspension is allowed to contact a sample in the presence of the above mentioned buffer solution to cause the immune agglutination reaction, and the turbidity generated by the insoluble carrier particle agglutination reaction is measured to determine the antigen or antibody in the sample (wherein R<1>, R<2> and R<3> may be the same or different, and independently represent a hydrogen atom, a hydroxyalkyl group or the like).

Description

technical field [0001] The present invention provides a reagent for turbidimetric immunoassay containing insoluble carrier particles such as latex, a method for turbidimetric immunoassay using insoluble carrier particles, and a kit for turbidimetric immunoassay containing insoluble carrier particles. More specifically, a reagent for turbidimetric immunoassay containing insoluble carrier particles that stabilizes absorbance and suppresses the action of plasma components related to the reaction that affects the measured value, and a turbidimetric immunoassay using insoluble carrier particles Assay method and kit for turbidimetric immunoassay containing insoluble carrier particles. technical background [0002] In the field of clinical examinations, latex is widely used in immunoassays for measuring antigens or antibodies in a sample. For example, Japanese Patent Laid-Open No. 10-253629 describes that in a buffer solution of pH 4.0 to 6.0, such as a phosphate citrate buffer so...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/58G01N33/53
CPCG01N33/54313G01N33/54393G01N33/543
Inventor 重信香代子小栗一人
Owner KYOWA MEDEX CO LTD
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