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Method for greatly improving chemiluminescence magnetic enzyme immunization sensitivity by depending on polystyrene nanoparticles

A polystyrene nano, chemiluminescence technology, applied in scientific instruments, material testing products, measuring devices, etc., can solve the problem of insufficient sensitivity, and achieve the effects of high sensitivity, fast detection speed and strong specificity

Active Publication Date: 2014-02-05
GETEIN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Another object of the present invention is to provide a detection kit, which can solve the problem that the sensitivity of the domestic kits in the prior art is not high enough in the detection process, and the detection sensitivity can be improved by at least 1-2 orders of magnitude after using the present invention

Method used

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  • Method for greatly improving chemiluminescence magnetic enzyme immunization sensitivity by depending on polystyrene nanoparticles
  • Method for greatly improving chemiluminescence magnetic enzyme immunization sensitivity by depending on polystyrene nanoparticles
  • Method for greatly improving chemiluminescence magnetic enzyme immunization sensitivity by depending on polystyrene nanoparticles

Examples

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Effect test

Embodiment 1

[0030] A kit for detecting cTnI, comprising sensitized nanoparticles 1, that is, polystyrene latex nanoparticle suspension covalently coupled with horseradish peroxidase-labeled mouse anti-human cTnI monoclonal antibody; sensitized magnetic particles 2 That is, the magnetic particle suspension covalently coupled with another mouse anti-human cTnI monoclonal antibody; 0.1mM chemiluminescence substrate luminol solution, 0.1mM luminescence enhancer p-iodophenol solution, and 3mM hydrogen peroxide solution. Add cTnI to the system containing the above two particles, and form a magnetic particle-antibody-antigen to be tested-enzyme-labeled antibody-polystyrene latex nanoparticle complex through the interaction of antibody-antigen-enzyme-labeled antibody, using magnetic separation technology Capture the complex, add the prepared luminol, p-iodophenol and hydrogen peroxide, and use a chemiluminescence detector to detect the luminescence intensity of the chemiluminescent substrate to c...

Embodiment 2

[0038] 1. Experimental equipment and reagents:

[0039] EnSpire multi-marker microplate detection system (PerkinElmer, USA), 96-well white plate (OptiPlate-96, PerkinElmer, USA), constant temperature water bath (HH-60, Changzhou Guohua Electric Co., Ltd.), constant temperature shaker (Taicang City Science and Education Equipment Factory), micropipette (Thermo Company), high-speed refrigerated centrifuge (TGL-16, Hunan Xiangyi Laboratory Instrument Development Co., Ltd.), 96-well plate magnetic separation rack (Tianjin Basele Chromatography Technology Development Center) . cTnI standard (BIO-RAD company), PBS-T buffer (KH 2 PO 4 3.35g / L, Na 2 HPO 4 -12H 2 O 17.9g / L, KCl 0.2g / L, NaCl 8.77g / L, Tween-200.5g / L, pH 6.96), the cTnI standard substance diluent is the PBS-T damping fluid containing 20% ​​bovine serum, detects the cTnI The preparation of the kit is the same as in Example 1.

[0040] 2. Experimental steps:

[0041] 2.1 Determination of the concentration of sensiti...

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Abstract

The invention discloses a method for greatly improving chemiluminescence magnetic enzyme immunization sensitivity by depending on polystyrene nanoparticles. The method comprises the following steps of: covalently coupling one enzyme-labeled antibody of an antigen to be detected to a polystyrene latex nanoparticle; covalently coupling the other one enzyme-labeled antibody of the antigen to be detected to a magnetic particle; adding the two particles into a system comprising the antigen to be detected; forming a magnetic particle-antibody-antigen to be detected-enzyme-labeled antibody-polystyrene latex nanoparticle composite through the interaction of antibody-antigen-enzyme-labeled antibody; capturing the composite by a magnetic separation technique; and calculating the concentration of the antigen to be detected by detecting luminous intensity of a chemiluminescence substrate by using a chemiluminescence detector. The method has the advantages of high specificity, high sensitivity, high detection speed and the like.

Description

technical field [0001] The invention belongs to the field of preventive and diagnostic medical testing, and relates to a method for greatly increasing the immunosensitivity of chemiluminescent magnetic enzymes by relying on polystyrene nanoparticles, in particular to a nanoparticle-dependent method for detecting trace amounts of cardiac troponin I in human blood A method for substantially increasing the sensitivity of chemiluminescent magnetic enzyme immunoassays. Background technique [0002] Cardiac troponin I (cTnI) is one of the subunits of the cardiac troponin complex, which has high myocardial specificity and sensitivity, and has been increasingly used in the diagnosis of myocardial infarction and angina pectoris. And differential diagnosis, and gradually have a tendency to replace myocardial enzymes. Under physiological conditions, the concentration of these proteins cannot be measured by existing immunological methods. During ischemia, the integrity of cell membran...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/546G01N33/577
Inventor 苏恩本叶森颜彬
Owner GETEIN BIOTECH
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