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Biomarker joint detection method based on peroxyoxalate ester chemiluminescence system and kit

A biomarker and chemiluminescence technology, which is applied in the field of chemiluminescence immunoassay biomarker diagnosis, can solve problems such as inability to detect multiple indicators, and achieve the effects of short reaction time, accurate results, and excellent linear range

Inactive Publication Date: 2021-05-11
湖南博奥瑞康生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Based on the reaction principle of chemiluminescent immunoassay detection products currently used in clinical testing, the use of the same luminescent marker results in a single reaction that can only detect one indicator, and cannot complete the detection of multiple indicators in a single reaction at the same time

Method used

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  • Biomarker joint detection method based on peroxyoxalate ester chemiluminescence system and kit
  • Biomarker joint detection method based on peroxyoxalate ester chemiluminescence system and kit
  • Biomarker joint detection method based on peroxyoxalate ester chemiluminescence system and kit

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Embodiment 1

[0044] A combined biomarker detection kit for detecting cTnI, NT-proBNP and D-dimer, comprising component 1, component 2, component 3 and component 4. The preparation method of each component is as follows:

[0045] 1. Preparation of component 1

[0046] 1) Labeling method of FAM-cTnI Ab1

[0047] ①Synthesis of single-stranded 5'-DIBO-DNA-FAM-3' modified with DIBO group at the 5' end and FAM at the 3' end;

[0048] ② Using SiteClick of ThermoFisher TM Antibody Azido Modification Kit treats cTnI Ab1 to modify the sugar chain on the Fc fragment of the antibody to generate a diazo group;

[0049] ③Reaction coupling 5'-DIBO-DNA-FAM-3' with diazotized cTnI Ab1 (N3-cTnI Ab1) to generate FAM-cTnI Ab1;

[0050]

[0051]

[0052] ④ After the reaction is completed, FAM-cTnIAb1 is obtained by purification with molecular sieves.

[0053] 2) Labeling method of Cy3-NT-proBNP Ab1

[0054] ①Synthesis of single-stranded 5'-DIBO-DNA-Cy3-3' modified with DIBO group at the 5' end and...

Embodiment 2

[0104] 1. Sample testing

[0105] 1) 20 cases of plasma samples were obtained from the Laboratory Department of Hunan Provincial People's Hospital. The plasma samples were drawn into EDTA (ethylenediaminetetraacetic acid) anticoagulant tubes through veins, and the upper layer of transparent liquid obtained by centrifugation did not contain blood cells.

[0106] 2) Use the high-sensitivity troponin-I assay kit, brain natriuretic peptide precursor assay kit, and D-dimer (D-dimer) assay kit to detect the above 20 samples respectively, and the detection steps are according to the instructions of each kit Just operate. The test results are shown in Table 4.

[0107] The high-sensitivity troponin-I assay kit adopts chemiluminescence microparticle immunoassay method, manufacturer: Abbott, Guoji injection 20172400287, batch number 17517UI01, valid until 2020-12-24;

[0108] Brain natriuretic peptide precursor detection kit adopts electrochemiluminescence method, manufacturer: Roche, G...

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Abstract

The invention provides a biomarker joint detection method based on a peroxyoxalate ester chemiluminescence system and a kit, and the method comprises the following steps: fully mixing a sample with a specific antibody solution marked with different fluorophores and a magnetic bead suspension coated with the specific antibody to generate a sandwich reaction; fixing and repeatedly rinsing by using a magnetic field, removing unreacted antibodies, adding components containing bisoxalate and hydrogen peroxide, fully reacting, detecting and recording the luminous intensity values of the reaction solution under different wavelengths, and calculating to obtain the concentration value of the biomarker in the sample. According to the method, multiple fluorescent agents are used for marking, multiple wavelength signals can be generated in one reaction so as to achieve the purpose of detecting multiple biomarkers at the same time, and meanwhile the method has the advantages of being high in sensitivity, accurate in result, easy to operate and short in reaction time.

Description

technical field [0001] The invention belongs to the technical field of chemiluminescent immunoassay biomarker diagnosis, and in particular relates to a biomarker combined detection method and kit based on a peroxyoxalate chemiluminescent system. Background technique [0002] Chemiluminescence immunoassay not only has the high sensitivity of chemiluminescence reaction, but also has the high specificity of immune system. Due to the advantages of fast analysis speed, wide linear range, no scattered light interference, no radioactive pollution, and simple equipment, this technology has been widely used in clinical diagnosis, life analysis, environmental science and other fields. [0003] Chemiluminescent immunoassay technology is mainly used for the detection and analysis of various antigens, haptens, antibodies, hormones, enzymes, fatty acids, vitamins and drugs. The most commonly used immunoassay technique developed later. [0004] The chemiluminescence immunoassay consists ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/533
CPCG01N21/64G01N33/533
Inventor 严竹林
Owner 湖南博奥瑞康生物科技有限公司