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Method Of Pretreating Specimen And Immunoassay Using The Same

a specimen and immunoassay technology, applied in the field of specimen pretreatment and immunoassay, can solve the problems of delay in identifying the actual cause of the disease, aggravate the symptoms of the disease, and the measurement method that operates using an immunoagglutination assay has not yet been used in practical use, so as to prevent non-specific reactions, improve the degree of seriousness of the disease of the patient, and improve the effect of simplicity

Inactive Publication Date: 2008-01-10
ARKRAY INC
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  • Summary
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AI Technical Summary

Benefits of technology

The present invention relates to a method for measuring a substance in a nasal secretion specimen using an immunoassay. By pretreating the specimen with a protease beforehand, non-specific reactions are prevented, allowing for accurate measurements. The method can be used with various types of proteases and can be applied to different types of nasal secretion specimens. The amount of influenza virus in the specimen can be measured quantitatively, providing valuable information for diagnosis and treatment.

Problems solved by technology

The technical problem addressed in this patent is the occurrence of non-specific reactions in immunochromatography assays or enzyme immunoassays when measuring specimens from nasal secretion due to the presence of macromolecular substances, such as glycoproteins. These reactions can cause false positive results, leading to inappropriate treatment and potentially aggravating the symptoms of the disease being tested. The inventors of the present invention conducted a study to develop a simple immunoagglutination assay for measuring specimens from nasal secretion without causing a large number of non-specific reactions.

Method used

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Examples

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Effect test

example 1

[0026] (Experiment 1)

Preparation of Latex Sensitized with Anti-Influenza A Antibody

[0027] 0.5 ml of an antibody solution (in 50 mM phosphate buffer solution, pH 7.4) prepared so as to contain a 0.8 mg / ml anti-influenza A mouse monoclonal antibody was mixed with 0.5 ml of 10% (w / v) polystyrene latex particles (Sekisui Chemical Co., Ltd., average particle diameter: 0.5 μm). The resultant mixture was incubated at 37° C. for 1 hour, thus sensitizing the latex particles with the antibody. To this mixture, BSA (Sigma, bovine serum albumin) was added so that its concentration was 1 wt %. Thereafter, the resultant mixture was incubated at 37° C. for 1 hour to perform blocking. The antibody-sensitized latex particles that had been subjected to the blocking were washed with a 50 mM phosphate buffer solution (pH 7.4) and then dispersed in a latex dispersion buffer solution (in 50 mM Tris-HCl buffer solution, pH 8.4, 0.1 wt % BSA, 0.1 wt % NaN3) so that its concentration was 0.1% (w / v). Thus...

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Abstract

The present invention provides a method of pretreating a specimen, which allows measurement according to an immunoassay to be carried out on a specimen from nasal secretion while preventing non-specific reactions. According to this method, the specimen from nasal secretion is treated with a protease beforehand and then an immunoassay is performed. As the protease, it is preferable to use semi-alkaline protease (EC 3.4.21.63). Furthermore, it is preferable that a substance to be pretreated by the pretreatment method according to the present invention is an influenza virus contained in the specimen from nasal secretion. The immunoassay preferably is an immunoagglutination assay. Examples of the immunoagglutination assay include a turbidimetric immunoassay, a latex turbidimetric immunoassay, and a latex agglutination assay that is performed on a slide glass.

Description

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Claims

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Application Information

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Owner ARKRAY INC
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