Islet cell antibody detection kit and preparation method thereof

A technology for detecting islet cells and antibodies, applied in the field of immunodiagnosis, can solve the problems of unstable accuracy and repeatability, and achieve the effects of improving specificity and detection speed, increasing luminescence rate, and improving sensitivity

Inactive Publication Date: 2018-12-28
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a detection kit for islet cell antibody and its preparation method in order to solve the problem of the accuracy and unstable repeatability of the existing ICA detection method

Method used

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  • Islet cell antibody detection kit and preparation method thereof
  • Islet cell antibody detection kit and preparation method thereof
  • Islet cell antibody detection kit and preparation method thereof

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preparation example Construction

[0037] The present invention also provides a preparation method of the islet cell antibody detection kit, comprising:

[0038] Step 1: Preparation of streptavidin magnetic particle suspension

[0039] After mixing the streptavidin magnetic particle solution and TBST solution, place it on a magnetic separator until the supernatant is free of turbidity, discard the supernatant, keep the magnetic particles, and prepare a solid phase reagent in the buffer after washing , to obtain streptavidin magnetic particle suspension;

[0040] The concentration of the streptavidin magnetic particle solution is preferably 50-100 mg / ml; the volume ratio of the streptavidin magnetic particle solution to the TBST solution is preferably (0.5-1): (5-10); The mixing time is preferably 10-15 minutes, the buffer is 50mM MES, 0.05% Tween-20, 0.05% Proclin300, pH6.5; the concentration of the solid-phase reagent is preferably 0.01% to 1 %; The source of the streptavidin magnetic particle solution is co...

Embodiment 1

[0048] Embodiment 1 The preparation method of the chemiluminescent immunoassay kit for islet cell antibody of the present invention comprises the following steps:

[0049] (1) Preparation of streptavidin magnetic particle suspension

[0050] Take 0.5 milliliter (50 mg) of streptavidin magnetic particle solution with a concentration of 100 mg / ml, add 10 mL of TBST solution and mix well for 10 minutes, then place it on a magnetic separator until the supernatant is free of turbidity, discard the supernatant, Keep the magnetic particles. After repeated washing for 3 times, a solid-phase reagent with a magnetic bead concentration of 0.05% was formulated in a buffer solution of 50mM MES, 0.05% Tween, 0.05% Proclin300, pH 6.5, and stored at 2-8°C.

[0051] (2) Preparation of acridinium ester-labeled anti-human IgG antibody

[0052] Put 500ug of anti-human IgG into the centrifuge tube, make sure that the anti-human IgG is at the bottom of the centrifuge tube (centrifuge at room temp...

Embodiment 2

[0059] Embodiment 2 The preparation method of the chemiluminescent immunoassay kit for islet cell antibody of the present invention comprises the following steps:

[0060] (1) Preparation of streptavidin magnetic particle suspension

[0061] Take 0.5 milliliter (50 mg) of streptavidin magnetic particle solution with a concentration of 100 mg / ml, add 10 mL of TBST solution and mix well for 15 minutes, then place it on a magnetic separator until the supernatant is free of turbidity, discard the supernatant, Keep the magnetic particles. After repeated washing for 3 times, a solid-phase reagent with a magnetic bead concentration of 0.05% was formulated in a buffer solution of 50mM MES, 0.05% Tween, 0.05% Proclin300, pH 6.5, and stored at 2-8°C.

[0062] (2) Preparation of acridinium ester-labeled anti-human IgG antibody

[0063] Put 500ug of anti-human IgG into the centrifuge tube, make sure that the anti-human IgG is at the bottom of the centrifuge tube (centrifuge at room temp...

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Abstract

The invention provides an islet cell antibody detection kit and preparation method thereof, and belongs to the technical field of immunodiagnosis. The kit comprises calibrating products, a streptavidin magnetic particle suspension, a chemiluminescent marker labeled anti-human IgG antibody, and a coupling labeled islet cell antigen. The invention further provides a preparation method of the islet cell antibody detection kit. The kit prepared by the preparation method has high detection sensitivity, is easy to operate, accurate and rapid in quantitative detection, and requires only 20 minutes todetect result; and paramagnetic particles are used as solid phase carriers, the specific surface area of the paramagnetic particles is large, so that the detection sensitivity is improved.

Description

technical field [0001] The invention belongs to the technical field of immunodiagnosis, and in particular relates to a detection kit for islet cell antibodies and a preparation method thereof. Background technique [0002] Type Ⅰ diabetes mellitus (T1DM) is caused by the autoimmune damage of pancreatic β cells in susceptible individuals, showing a gradual decrease or lack of insulin (Ins) secretion. Islet cell antibody (ICA) is one of the antibodies that can be detected in the serum of patients with T1DM when the humoral immunity is abnormal, and it is a serological marker of slowly progressive β-cell injury. Pancreatic islets are composed of a variety of endocrine cells. β cells are distributed in the center of the islets and secrete insulin. α cells located around the islets are also involved in the regulation of insulin secretion. ICAs can be divided into islet cell cytoplasmic antibodies (ICA) and islet cell surface antibodies (ICSA) according to their binding sites on ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/74
Inventor 李冬梅高智玲高威孙成艳何浩会
Owner DIRUI MEDICAL TECH CO LTD
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