JPH0634014B2 - Colored particles for immunodiagnosis - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a colored particle for immunodiagnosis which is colored without deterioration of diagnostic accuracy.

[Prior Art] Conventionally, particles of synthetic polymers such as polystyrene have been used as immunodiagnostic particles. However, synthetic polymers are essentially colorless or light-colored. It cannot meet the demand for improving the accuracy of diagnosis by using or coloring. Therefore, various attempts have been made to color the particles for immunodiagnosis, but in most cases, the accuracy of diagnosis has been remarkably lowered.

[Object] Therefore, an object of the present invention is to provide colored particles for immunodiagnosis without deteriorating the accuracy of diagnosis.

[Structure of the Invention] As a result of various studies on the coloring method of the particles for immunodiagnosis in view of the above-mentioned demands and problems, the present inventors have found that if the particles are colored with a specific dye, the diagnostic accuracy decreases. Without,
Rather, they found that they could improve, and completed the present invention with the recognition.

That is, the particle for immunodiagnosis of the present invention is a homopolymer of styrene having a particle size of 0.05 to 100 μm when mixed with at least one dye solution selected from azo or quinone oil-soluble dyes. The present invention is characterized in that immunodiagnostic particles made of a copolymer are colored.

The particle for immunodiagnosis used in the present invention has a particle size of 0.05 to 100 μm and is used in a state of being dispersed in water.

The immunodiagnostic particles having a particle size of 0.05 to 100 μm can be produced by an emulsion polymerization method, a seed polymerization method, a precipitation polymerization method, a suspension polymerization method, a redispersion method or the like. Coloring with a dye is performed by mixing a dye solution and particles.

As the material of such particles, homopolymers and copolymers of styrene can be used, and polystyrene, a polymer copolymerizable with styrene and having a carboxyl group, a hydroxyl group, an amino group, an aldehyde group, a sulfonic acid group, etc. It may be a copolymer with a monomer, or one obtained by introducing a carboxyl group, a hydroxyl group, an amino group, an aldehyde group, a sulfonic acid group or the like into polystyrene by a chemical reaction or plasma treatment.

An azo or quinone oil soluble dye is used as the dye. Specifically, solvent yellow 2, solvent yellow 14, solvent orange 1, solvent red 2
3, azo oil-soluble dyes such as Solvent Red 1, quinone oil-soluble dyes such as Oil Red 9, Solvent Violet 13 and Solvent Blue 12. These dyes are soluble in a solvent that dissolves or swells polymer particles, but not in water. This allows the particles to be dyed by mixing the aqueous dispersion of particles, ie the latex and the dye solution.

Other dyes can be used as dyes, but often cause agglomeration of particles, decolorization of dyes, or deterioration of diagnostic accuracy. Solvent Yellow 1 is a preferred dye.
4, Solvent Red 1, Solvent Blue 12, and mixtures thereof.

The amount of dye used is about 0.1 to 100 ppm of the polymer.

By treating the colored particles for immunodiagnosis colored as described above with serum or serum protein, the diagnostic accuracy can be further improved by treating serum or serum protein before sensitizing the particles with an antibody or an antigen. Or even after sensitization.

As the serum, bovine serum, calf serum, rabbit serum, sheep serum, chicken serum, human serum, etc. are used, and usually diluted with a phosphate buffer solution 2 to 20 times before use. Examples of serum proteins include albumin, globulin, fibroin, etc., with albumin being preferred. Serum protein is used as a 0.1-10 wt% phosphate buffer solution.

As a treatment method, particles or an aqueous dispersion of particles and a serum diluent or a serum protein solution are used at room temperature, for example, 2
After mixing for an hour, the supernatant can be discarded and washed with phosphate buffer.

The immunoreactive substance carried by the colored particles for immunodiagnosis according to the present invention includes hepatitis B virus surface antigen (H
Bs antigen), anti-HBs antibody, human chorionic gonadotropin (HCG antigen), anti-HCG antibody, immunoglobulin G,
Examples include mycoplasma antigens, nucleic acids, nucleoproteins, estrogens, anti-estrogen antibodies, and the like.

EXAMPLES Next, the present invention will be described in more detail with reference to Examples.
Of course, it is not limited to these.

Example 1 To 100 g of a styrene-methacrylic acid copolymer latex having a solid content concentration of 10% and an average particle size of 0.7 μm, 2 ml of a 0.1% toluene solution of a dye was gradually added with stirring at room temperature. Stir for 2 hours at 35 ° C. When toluene was removed by steam distillation, a latex of colored particles was obtained. Solvent Yellow 14 (azo oil soluble dye), Solvent Red 1 (azo oil soluble dye), Solvent Blue 12 (quinone oil soluble dye) and Oil Red 9 (quinone oil soluble dye) were used as dyes. The obtained colored latex was sensitized with an antigen of rheumatoid factor, and the detection sensitivity of the rheumatoid factor by the slide test was determined. As a result, all were similar to the uncolored latex.

Example 2 100 g of a latex of a styrene-tribromophenyl acrylate-methacrylic acid copolymer having a solid content concentration of 10% and an average particle diameter of 1.4 μm was added to 0.01 g of the dye shown in Table 1 at room temperature.
5 ml of a% chloroform solution was gradually added while stirring, and after the addition was completed, the mixture was stirred at 50 ° C. for 2 hours. Blowing steam to remove chloroform gave a latex with dispersed particles colored.

The obtained colored latex was sensitized with an antigen of rheumatoid factor, and the detection sensitivity of the rheumatoid factor was measured by the microtiter method. The results are shown in Table 1.

Comparative Example 1 In Example 1, as a dye, paranyl lumina red G (methine dye), Michelen Brilliant Pink R
(Indigo dye), copper phthalocyanine (phthalocyanine dye) or rose bengal methylene blue (xanthene dye) was used in the same manner as in Example 1.

As a result, when any of the dyes was used, the agglutination image was weaker than that of the uncolored latex and the detection sensitivity was poor.

[Effects] As described above, according to the present invention, an immunodiagnosis comprising a styrene homopolymer or a copolymer using at least one dye solution selected from azo or quinone oil-soluble dyes. By coloring the particles for use, there is an advantage that the diagnostic accuracy can be improved without lowering the diagnostic accuracy.

Claims (1)

[Claims] 1. An immunodiagnosis comprising a homopolymer or copolymer of styrene having a particle size of 0.05 to 100 μm when mixed with at least one dye solution selected from azo or quinone oil soluble dyes. Colored particles for immunodiagnosis, characterized in that the particles for coloring are colored.