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Plate type magnetic microparticle chemiluminescence analysis kit for detecting leptin antigen

A technology of magnetic particles and chemiluminescence, which is applied in chemiluminescence/bioluminescence, analysis of materials through chemical reactions, etc., can solve the problems of high price, harm to human body and environment, and meet the requirements of pretreatment Low, cheap, simple process effect

Active Publication Date: 2017-02-15
上海英基生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Currently, the immunological methods used to detect leptin antigen mainly include radioimmunoassay (RIA) and enzyme immunoassay (enzyme immunoassay, EIA). The markers of the RIA kit are radioactive, which is harmful to human body and environment; EIA Most of the kits are imported from abroad, and the price is expensive

Method used

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  • Plate type magnetic microparticle chemiluminescence analysis kit for detecting leptin antigen
  • Plate type magnetic microparticle chemiluminescence analysis kit for detecting leptin antigen
  • Plate type magnetic microparticle chemiluminescence analysis kit for detecting leptin antigen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] 1. The magnetic microparticle plate chemiluminescent immunoassay kit for detecting leptin antigen includes:

[0024] (1) Commercially available magnetic microparticles coated with anti-FITC antibody: the particle size is 2-3 μm, the concentration used is 5-10 mg / mL, 20 mL / bottle, 1 bottle;

[0025] (2) FITC-labeled leptin monoclonal antibody (capture antibody) and horseradish peroxidase-labeled leptin polyclonal antibody (paired antibody) are mixed as a marker mixture: FITC-labeled capture with a volume ratio of 1:1 Antibody working solution and enzyme-labeled paired antibody working solution are mixed, 15mL / bottle, 1 bottle, in which the FITC-labeled antibody solution is diluted to 1.25μg / mL with 20% calf serum, and the dilution used for the enzyme working solution labeled with horseradish enzyme The solution is a biological preservative solution containing 15% glycerin and 0.05% and diluted to 1 / 2000;

[0026] (3) Leptin standard solution: use 50% horse serum to dilu...

Embodiment 2

[0053] Example 2 Kit Precision, Accuracy, Soundness, Sensitivity and Stability Experiments

[0054] 1. Kit precision test standard product precision experiment

[0055] (1) Three batches of the kits prepared in Example 1 were taken respectively for the precision test, and 10 kits were taken from each batch. The leptin standard substance of 3.125 ng / mL was measured 6 times with the kit extracted in Example 1, and the coefficient of variation of the measured concentration was calculated. The measurement results of the three batches of kits in Example 1 are shown in Table 1, and the results show that the coefficient of variation is between 4.3-8.0%.

[0056] Table 1 Leptin standard repeatability experiment

[0057]

[0058] (2) Sample repeatability experiment Take 1 part of normal human serum and add leptin standard to 3.0 ng / mL. Three different batches of the kits in Example 1 were taken, and the samples were repeatedly measured 6 times, and the coefficient of variation wa...

Embodiment 3

[0068] The magnetic microparticle plate chemiluminescence immunoassay kit for detecting leptin antigen contains Chinese crabgrass extract, which is set in a vial of magnetic microparticles coated with anti-FITC antibody, and the content is 1-3 mg / mL. The addition of Chinese crabgrass extract The stability of the kit was increased. After the kit of Example 3 was subjected to accelerated experiments at 37°C for 15 days and 27 days, the maximum and minimum luminous intensity of the kit and the recovery rate of the analyte were measured, indicating that the indicators of Example 1 within the normal range. The main components of the kit in Example 3: marker working solution, standard substance, luminescent substrate solution, and magnetic particle working solution were followed up at 2-8°C for 12 months, and the results showed that all indicators met the requirements. Considering the occurrence of the freezing of the kit, the kit of Example 3 was frozen at -20°C for 10 days, and th...

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Abstract

The invention discloses a plate type magnetic microparticle chemiluminescence analysis kit for detecting a leptin antigen, and belongs to the technical field of biological detection. The kit comprises a magnetic microparticle coated with an antibody against fluorescein isothiocyanate, a liquid mixture of a marker, a leptin antigen standard substance, a concentrated washing solution and a luminescent substrate solution. The plate type magnetic microparticle chemiluminescence enzyme immunoassay kit is simple in structure, convenient to use, low in price, convenient to carry, and compared with radioimnuno assay kits and enzyme linked immunosorbent assay kits in the market, the plate type magnetic microparticle chemiluminescence enzyme immunoassay kit has higher sensitivity and wider linear range, effectively avoids bending hook effect, does not need sample dilution, and is suitable for qualitative in quantitative screening of large quantities of samples.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a plate-type magnetic microparticle chemiluminescence analysis kit for detecting leptin antigen. Background technique [0002] Leptin is a protein hormone secreted by fat cells, with a total length of 146 amino acids. Specific receptors (OB-R) and corresponding signal transduction systems play a role. After leptin enters the blood circulation, it will participate in the regulation of sugar, fat and energy metabolism. Many recent studies at home and abroad have found that leptin levels will change under many physiological and pathological conditions, which has very important clinical guiding value for the occurrence, development, prognosis and efficacy testing of many diseases related to energy metabolism. For example: obesity and increased leptin levels and the disappearance of leptin circadian rhythm are mutual causality, leptin levels in the body have...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/74G01N33/577G01N33/543G01N33/533G01N33/535G01N21/76
CPCG01N21/76G01N33/533G01N33/535G01N33/54326G01N33/577G01N33/74
Inventor 杨凌波
Owner 上海英基生物科技有限公司
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