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Detection kit

A detection kit and reagent technology, applied in the field of medical devices, can solve the problems of inability to visually evaluate antigen/antibody levels, low accuracy, and susceptibility to interference, and achieve the effects of reducing the cost of medical institutions, rapid detection, and simple color development

Active Publication Date: 2019-10-18
深圳海思安生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the limitations of the solid-phase reaction system, this method has poor sensitivity, is susceptible to interference, and has low accuracy.
In addition, although this method does not need to dilute the sample and can reflect the actual level of the body, it can only be used for qualitative detection and cannot make an intuitive evaluation of the level of antigen / antibody

Method used

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preparation example Construction

[0063] The washing buffer used in the preparation of reagent 1 is a buffer commonly used in biological experiments, and can include PBS buffer, Tris, Tween-20, MES, SDS, etc., preferably 20-100nm MES, 0.001-0.1% SDS The buffer solution, the pH is between 6 and 10. The activation buffer is a buffer commonly used in biological experiments, and can contain EDC, NHS, SDS, etc., preferably a buffer containing 100-500nm EDC and 100-500nm NHS, and the quencher is a commonly used quencher in biological experiments. Contains 2-mercaptoethanol, glycine, ethanolamine, etc., preferably one or both of ethanolamine and 2-mercaptoethanol.

[0064] Reagent 2: Reagent 2 contains non-magnetic microspheres with multiple colors, wherein the surface of the non-magnetic microspheres of the first color a is bound with substance B, and the surface of the microspheres of other colors is not bound with substance B. These non-magnetic microspheres are suspended in blocking solution and usually stored a...

preparation Embodiment 1

[0105] 1) Preparation of Reagent 1

[0106] Take 20 μL of the magnetic particle stock solution (particle size 500 nm, 10% w / v) into a 2 mL test tube, add 1 mL of MES washing buffer and mix thoroughly, place on a magnetic separation rack, and suck out the supernatant after the magnetic particles are precipitated. Repeat the above washing step 2-3 times, after washing, add 1 mL of MES washing buffer to resuspend the magnetic particles. Then add 12 μL of freshly prepared LEDC and 120 μL of NHS activation buffer, vortex and mix on a roller mixer at 100 rpm for 30 minutes at room temperature, place on a magnetic separation rack and suck out the supernatant after the magnetic particles are precipitated. Add 1mL MES washing buffer to wash the magnetic particles, and repeat 2-3 times. After washing, add 850 μL MES washing buffer to resuspend the magnetic particles.

[0107] Dilute dsDNA to a concentration of 100 μg / mL with MES washing buffer, add 150 μL to the above suspension, mix ...

preparation Embodiment 2

[0120] Method is as described in Preparation Example 1, the difference is:

[0121] The particle size of the magnetic particles used was 5000nm, the dosage was 100 μL, and the concentration of the coupled dsDNA antigen was 500 μg / mL.

[0122] The carboxyl latex microspheres used had a particle size of 2000 nm, the amount of blue microspheres was 100 μL, the concentration of the coupled anti-dsDNA antibody was 500 μg / mL, and the amount of red microspheres was 50 μL.

[0123] The volume ratio of blue microspheres and red microspheres is 4:1,

[0124] The blocking solution is composed of 0.5M glycine, 0.9% NaCl, 2% BSA, 0.5% Tween-20, 0.5% Proclin 300, and the pH is 10.

[0125] Take 29 serum samples with clinically confirmed results, and use the kits prepared in Preparation Examples 1 and 2 to detect them respectively.

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Abstract

The invention provides a detection kit. The kit is used for detecting a substance to be detected in a sample to be tested. The detection kit contains two liquid reagents of: a reagent 1 and a reagent2; the reagent 1 comprises magnetic micro-particles, and the surface of the micro-particles is combined with a substance A which can specifically bind to the substance to be detected in the sample; the reagent 2 comprises non-magnetic microspheres with one or more colors, wherein the surface of the microspheres of a first color is bonded with a substance B, the surface of the microspheres of the remaining colors is not bound with the substance B, and the substance B can specifically bind to the substance A, which is the same as or different from the substance to be detected. When the detectionis carried out using the kit provided by invention, the level of the substance to be detected can be quantitatively or semi-quantitatively determined according to the color of the reaction solution.The solution color more tends to the first color, it is indicated that the level of the substance to be detected is higher. The detection kit provided by the invention can obtain the detection resultonly by the naked eye, which can not only achieve rapid detection, but also ensure that the detection result is intuitive and accurate; therefore, the detection kit can be effectively used in health and medical institutions, especially in health and medical institutions.

Description

technical field [0001] The invention belongs to the field of medical devices, and in particular relates to an in vitro diagnostic reagent, in particular to an in vitro detection reagent for assisting disease diagnosis. Background technique [0002] Autoimmune diseases are characterized by the appearance of autoreactive lymphocytes and autoantibodies against self-antigens, immunoglobulins in affected tissues. Systemic Lupus Erythematosus (SLE) is a typical representative of autoimmune diseases, often with multi-system damage and the production of various autoantibodies. The most prominent manifestation in its pathogenesis is hyperfunction or over-activation of B lymphocytes Instead, large amounts of polyclonal immunoglobulins and autoantibodies directed against self-tissues are produced spontaneously. The skin is the most common organ involved, and skin damage is the primary reason for patients to see a doctor, and it is also the most suggestive clinical diagnosis basis and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/533G01N33/543G01N33/569G01N21/78G01N33/68
CPCG01N33/535G01N33/533G01N33/54326G01N33/56983G01N33/56927G01N33/56933G01N21/78G01N33/68Y02A50/30
Inventor 牛海涛凌健东许爱红管志平杨雪
Owner 深圳海思安生物技术有限公司
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