Composition for enzyme linked immunosorbent assay kit, anti-nuclear antibody spectrum detection kit and preparation method thereof

Abstract

The invention relates to the technical field of enzyme linked immune, and discloses a composition for an enzyme linked immunosorbent assay kit, an anti-nuclear antibody spectrum detection kit and a preparation method thereof. The composition comprises sealing liquid and ELISA diluents; the sealing liquid contains casein, saccharose, PBS, mannitol, Tween20, lycine and sodium azide; the ELISA diluents contain PBS, casein, sodium p-hydroxybenzoate, PEG, lycine and Proclin300. By starting from the sealing liquid and the ELISA diluents of the enzyme linked immunosorbent assay kit, the proper ingredients are selected, so that the enzyme linked immunosorbent assay kit can maintain the detection stability for a long time. Meanwhile, the anti-nuclear antibody spectrum detection kit prepared from the composition has higher stability; the guarantee period is 2 years or more.

Description

technical field

[0001] The invention relates to the technical field of ELISA, in particular to a composition used in an ELISA kit, an antinuclear antibody spectrum detection kit and a preparation method thereof. Background technique

[0002] Antinuclear antibody (ANA) is an autoantibody with various nuclear components, a general term for autoantibodies that target the nuclear component of eukaryotic cells, and can be used for screening of various autoimmune diseases. Autoimmune disease (AID) refers to a series of pathological immune responses produced by the body's immune effects or immune effector molecules against its own tissues or cells, resulting in damage to its own tissues and organs. Its pathogenesis is very complex and diverse. Changes often involve the skin, serosa, joints, kidneys, and the central nervous system. Common diseases include systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), Sjogren's syndrome (SS), mixed connective tissue disease (MCTD), ...

Examples

Embodiment 1

[0038] Embodiment 1: Preparation of antinuclear antibody spectrum detection kit of the present invention

[0039] 1. Coating of nuclear component antigens and related proteins

[0040] The PC, NC, S1, S2, S3, S4, S5, and EC points in the protein chip array are coated with 2ug / ml, 0.01ug / ml, 0.1ug / ml, 0.5ug / ml, 2ug / ml, 4ug respectively / ml, 8ug / ml, 2ug / ml of human IgG, the dilution buffer is CB buffer at pH9.6 (which contains 2.5% PEG4000, 5% trehalose, 0.05% Proclin300, and 15% glycerol) .

[0041] SC dots are coated with 2ug / ml goat anti-human IgG antibody, and the dilution buffer is CB buffer at pH 9.6.

[0042] Loc dots are coated with 2ug / ml human IgG, and the dilution buffer is CB buffer at pH 9.6.

[0043] The 7 antigens of dsDNA, Sm, U1RNP, Scl-70, Jo-1, histone, and nucleosome were respectively treated with 0.01M PBS buffer of pH7.4-PH7.6 (containing 0.5% PVP, 5% Trehalose, 0.05% Proclin300, 0.02% 2-hydroxy-β-cyclodextrin) were diluted, and the final concentrations...

Embodiment 2

[0068] Embodiment 2: Stability detection of the kit of the present invention

[0069] Control kit: prepared according to the method of Example 1, the difference is that the blocking solution is a conventional PBS buffer solution containing 3% BSA and 0.05% Proclin300, pH7.4; the enzyme label diluent uses the enzyme in the prior art CN105021811A Standard diluent: 0.46% tris, 5% goat serum, 0.8% NaCl, 0.1% Tween 20, 0.04% EDTA, 0.5% sodium paraben, 0.1% Proclin300, the balance is deionized water.

[0070] Test kit: embodiment 1 kit;

[0071] Detection method: place the two kits at room temperature (18-28°C) and low temperature (2-8°C) for a period of time, then use the same serum to detect according to the detection method in Example 1, and count the signal value of the instrument. The results are shown in Table 2-5.

[0072] 1. The stability data of the kit in Example 1 at low temperature

[0073] Table 2

[0074]

[0075]

[0076]

[0077] As can be seen from Table...

technical field

[0001] The invention relates to the technical field of ELISA, in particular to a composition used in an ELISA kit, an antinuclear antibody spectrum detection kit and a preparation method thereof. Background technique

[0002] Antinuclear antibody (ANA) is an autoantibody with various nuclear components, a general term for autoantibodies that target the nuclear component of eukaryotic cells, and can be used for screening of various autoimmune diseases. Autoimmune disease (AID) refers to a series of pathological immune responses produced by the body's immune effects or immune effector molecules against its own tissues or cells, resulting in damage to its own tissues and organs. Its pathogenesis is very complex and diverse. Changes often involve the skin, serosa, joints, kidneys, and the central nervous system. Common diseases include systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), Sjogren's syndrome (SS), mixed connective tissue disease (MCTD), ...

Examples

Embodiment 1

[0038] Embodiment 1: Preparation of antinuclear antibody spectrum detection kit of the present invention

[0039] 1. Coating of nuclear component antigens and related proteins

[0040] The PC, NC, S1, S2, S3, S4, S5, and EC points in the protein chip array are coated with 2ug / ml, 0.01ug / ml, 0.1ug / ml, 0.5ug / ml, 2ug / ml, 4ug respectively / ml, 8ug / ml, 2ug / ml of human IgG, the dilution buffer is CB buffer at pH9.6 (which contains 2.5% PEG4000, 5% trehalose, 0.05% Proclin300, and 15% glycerol) .

[0041] SC dots are coated with 2ug / ml goat anti-human IgG antibody, and the dilution buffer is CB buffer at pH 9.6.

[0042] Loc dots are coated with 2ug / ml human IgG, and the dilution buffer is CB buffer at pH 9.6.

[0043] The 7 antigens of dsDNA, Sm, U1RNP, Scl-70, Jo-1, histone, and nucleosome were respectively treated with 0.01M PBS buffer of pH7.4-PH7.6 (containing 0.5% PVP, 5% Trehalose, 0.05% Proclin300, 0.02% 2-hydroxy-β-cyclodextrin) were diluted, and the final concentrations...

Embodiment 2

[0068] Embodiment 2: Stability detection of the kit of the present invention

[0069] Control kit: prepared according to the method of Example 1, the difference is that the blocking solution is a conventional PBS buffer solution containing 3% BSA and 0.05% Proclin300, pH7.4; the enzyme label diluent uses the enzyme in the prior art CN105021811A Standard diluent: 0.46% tris, 5% goat serum, 0.8% NaCl, 0.1% Tween 20, 0.04% EDTA, 0.5% sodium paraben, 0.1% Proclin300, the balance is deionized water.

[0070] Test kit: embodiment 1 kit;

[0071] Detection method: place the two kits at room temperature (18-28°C) and low temperature (2-8°C) for a period of time, then use the same serum to detect according to the detection method in Example 1, and count the signal value of the instrument. The results are shown in Table 2-5.

[0072] 1. The stability data of the kit in Example 1 at low temperature

[0073] Table 2

[0074]

[0075]

[0076]

[0077] As can be seen from Table...

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