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High-sensitivity chemiluminescence immunoassay kit, and preparation method and application thereof

A chemiluminescence immunoassay and analytical reagent technology, which is applied in the field of high-sensitivity chemiluminescence immunoassay kits and its preparation, can solve the problems of low sensitivity and achieve the effects of improved sensitivity, good repeatability and strong specificity

Inactive Publication Date: 2018-07-27
南京仁迈生物科技有限公司
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a high-sensitivity chemiluminescence immunoassay kit, which is based on a high-sensitivity chemiluminescence immunoassay method based on luminescent substance-labeled streptavidin cycle amplification, which can improve sensitivity and solve the problem of existing chemiluminescence immunoassay reagents. The problem of low box sensitivity

Method used

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  • High-sensitivity chemiluminescence immunoassay kit, and preparation method and application thereof
  • High-sensitivity chemiluminescence immunoassay kit, and preparation method and application thereof
  • High-sensitivity chemiluminescence immunoassay kit, and preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: sandwich method comparative experiment (Free PSA)

[0042] 1. Solid phase carrier directly coated with antibody

[0043] 1. Take the magnetic bead stock solution, wash the magnetic beads twice with 50mM MES buffer solution, and then resuspend in the buffer solution. A certain concentration of EDC ready-made solution was added thereto, and activated at 37° C. for 0.5 hours. After activation, wash with MES buffer solution three times, and then resuspend in reaction buffer solution.

[0044] 2. Add the antibody used and react at 37°C for 3 hours. After the reaction is completed, wash with MES buffer solution three times, and finally resuspend in magnetic bead storage solution.

[0045] 2. Analysis steps

[0046] Concentration gradients were prepared with PBS buffer solution: 0.01, 0.1, 0.2, 0.4, 1.0, 2.0, 4.0, 10.0, 20.0, 30.0, 40.0, 50.0ng / mL Free PSA standard. In a cuvette, add 20 μL of magnetic beads coated with one strain of Free PSA monoclonal antibod...

Embodiment 2

[0054] Embodiment 2: competition method and comparative test (E2)

[0055] 1. Solid phase carrier directly coated with antigen

[0056] 1. Take the original solution of magnetic beads, take the reaction buffer solution to wash the magnetic beads, and then resuspend in the buffer solution. A certain concentration of EDC ready-made solution was added thereto, and activated at 37° C. for 0.5 hours. After activation, wash three times with magnetic bead washing solution, and then resuspend in reaction buffer solution.

[0057] 2. Add the antigen used and react at 37°C for 3 hours. After the reaction is completed, wash with magnetic bead washing solution three times, and finally resuspend in magnetic bead storage solution.

[0058] 2. Analysis steps

[0059] Concentration gradients were prepared with PBS buffer solution: 10, 20, 40, 80, 160, 320, 640, 1200, 2500, 3000, 4000, 4800pg / mL E2 standard. Add 30 μL E2 standard, 20 μL E2 displacer, 50 μL biotin-labeled E2 monoclonal ant...

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Abstract

The invention provides a high-sensitivity chemiluminescence immunoassay kit, and a preparation method and application thereof, belonging to the technical field of clinical immunoassay. The high-sensitivity chemiluminescence immunoassay kit comprises a solid-phase vector directly coated with a capture molecule, a biotin-labeled marker molecule and luminescent substance-labeled streptavidin, whereinthe marker molecule is capable of binding to a target analyte in a to-be-detected sample, and the capture molecule is capable of specifically binding to the target analyte in the to-be-detected sample or is an analog of the target analyte in the to-be-detected sample; or the marker molecule is capable of competitively binding to the target analyte in the to-be-detected sample, and the capture molecule is capable of specifically binding to the target analyte in the to-be-detected sample. The kit provided by the invention is based on the structural characteristics of streptavidin; per mole of tetramer molecules can bind to four moles of biotin molecules, and strong affinity are formed between the tetramer molecules and the biotin molecules; and a cyclic amplification reaction process is constructed to realize signal amplification, so the sensitivity of the kit is improved.

Description

technical field [0001] The invention belongs to the technical field of clinical immunoassay, and in particular relates to a high-sensitivity chemiluminescence immunoassay kit and its preparation method and application. Background technique [0002] Chemiluminescence immunoassay (CLIA) is a new immunoassay technology established by combining luminescence analysis technology and immunoassay technology. Advantages of operation. In addition, it has the characteristics of environmental protection, long shelf life of reagents, high stability, and easy automation, making it one of the most promising methods in non-radioactive immunoassays. Nevertheless, with the popularization of chemiluminescence immunoassay technology and the increasing clinical demand for the sensitivity of molecules to be detected, there is an urgent need for chemiluminescence immunoassay technology with higher sensitivity. [0003] The patent discloses an analysis method using acridinium esters to label anti...

Claims

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Application Information

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IPC IPC(8): G01N33/532G01N33/543
CPCG01N33/532G01N33/54326
Inventor 金晶王西龙郭智慧
Owner 南京仁迈生物科技有限公司
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