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Preparation method of cTnI, NT-proBNP and D-dimer chest pain triple colloidal gold immunostrip

A technology of colloidal gold and test strips, applied in the field of medical testing, can solve problems such as early warning, diagnosis and treatment of unfavorable heart diseases, and achieve significant economic and social benefits, effective diagnosis and identification, and simple processes

Pending Publication Date: 2018-12-21
HENAN ACAD OF SCI INST OF BIOLOGY LIABILITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, the detection methods for cTnI, NT-proBNP and D-dimer include enzyme-linked immunoassay, fluorescent immunoassay, chemiluminescence, immunoturbidimetry, and colloidal gold immunochromatography, etc., but the detection products and methods are mainly It is limited to the detection of one or two of them, and there are very few methods for simultaneous detection of these three items (especially colloidal gold immunoassay), which is not conducive to more rapid, comprehensive and specific detection of various heart diseases. It is the occurrence, development and whole process of early warning, diagnosis and treatment of ACS

Method used

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  • Preparation method of cTnI, NT-proBNP and D-dimer chest pain triple colloidal gold immunostrip
  • Preparation method of cTnI, NT-proBNP and D-dimer chest pain triple colloidal gold immunostrip
  • Preparation method of cTnI, NT-proBNP and D-dimer chest pain triple colloidal gold immunostrip

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Embodiment 1

[0026] A method for preparing a cTnI, NT-proBNP and D-dimer chest pain triple colloidal gold immune test strip of the present invention, in specific implementation:

[0027] The preparation of the monoclonal antibody: by volume: take 1 part each of the mouse ascites containing the first anti-cTnI protein conjugate monoclonal antibody, the second anti-cTnI protein conjugate monoclonal antibody mouse ascites, Mouse ascites of the first anti-NT-proBNP protein conjugate monoclonal antibody, mouse ascites of the second anti-NT-proBNP protein conjugate monoclonal antibody, first anti-D-dimer protein conjugate monoclonal antibody Mouse ascites and mouse ascites with the second anti-D-dimer protein conjugate monoclonal antibody were mixed with 2 parts of 60mM acetate buffer, and octanoic acid was added drop by drop under stirring at room temperature at 18°C, and 30 μL of octanoic acid was added to each 1 mL of mouse ascites After mixing, place at room temperature for 28 minutes, centr...

Embodiment 2

[0033] A method for preparing a cTnI, NT-proBNP and D-dimer chest pain triple colloidal gold immune test strip of the present invention, in specific implementation:

[0034] Said preparation of monoclonal antibodies: by volume, take 1 part of mouse ascites containing the first anti-cTnI protein conjugate monoclonal antibody, mouse ascites containing the second anti-cTnI protein conjugate monoclonal antibody, and the second mouse ascites containing anti-cTnI protein conjugate monoclonal antibody. Primary anti-NT-proBNP protein conjugate monoclonal antibody mouse ascites, second anti-NT-proBNP protein conjugate monoclonal antibody mouse ascites, primary anti-D-dimer protein conjugate monoclonal antibody small Rat ascites and mouse ascites of the second anti-D-dimer protein conjugate monoclonal antibody were mixed with 2 parts of 60mM acetate buffer, and octanoic acid was added dropwise under stirring at 22°C, and 33 μL of octanoic acid was added to each 1 mL of mouse ascites, and...

Embodiment 3

[0040] A method for preparing a cTnI, NT-proBNP and D-dimer chest pain triple colloidal gold immune test strip of the present invention, in specific implementation:

[0041] The preparation of monoclonal antibodies: by volume, respectively take 1 part of mouse ascites containing the first anti-cTnI protein conjugate monoclonal antibody, mouse ascites containing the second anti-cTnI protein conjugate monoclonal antibody, Mouse ascites of the first anti-NT-proBNP protein conjugate monoclonal antibody, mouse ascites of the second anti-NT-proBNP protein conjugate monoclonal antibody, first anti-D-dimer protein conjugate monoclonal antibody Mouse ascites and the mouse ascites of the second anti-D-dimer protein conjugate monoclonal antibody were mixed with 2 parts of 60mM acetate buffer, and octanoic acid was added dropwise under stirring at 25°C, and 35 μL of octanoic acid was added to each 1 mL of mouse ascites, After mixing, place at room temperature for 32 minutes, centrifuge at...

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Abstract

The invention relates to a preparation method of a cTnI, NT-proBNP and D-dimer chest pain triple colloidal gold immunostrip. The detection problem of cTnI, NT-proBNP and D-dimer can be effectively solved. The preparation method comprises the following steps: respectively preparing an anti-cTnI protein conjugate monoclonal antibody, an anti-NT-proBNP protein conjugate monoclonal antibody and an anti-D-dimer protein conjugate monoclonal antibody for colloidal gold marking and a coating detection line; respectively preparing colloidal gold markers of the anti-cTnI protein conjugate monoclonal antibody, the anti-NT-proBNP protein conjugate monoclonal antibody and the anti-D-dimer protein conjugate monoclonal antibody, drying and curing the colloidal gold markers, sequentially coating the anti-cTnI protein conjugate monoclonal antibody, the anti-NT-proBNP protein conjugate monoclonal antibody and the anti-D-dimer protein conjugate monoclonal antibody for the coating detection line on a nitrocellulose membrane from left to right to form a detection line, and coating an anti-mouse immunoglobulin antibody on the nitrocellulose membrane to form a quality control line of the coating membraneso as to prepare the cTnI, NT-proBNP and D-dimer chest pain triple colloidal gold immunostrip. The preparation method provided by the invention is simple in process and is novel and unique, and various acute chest pains can be diagnosed and identified quickly, conveniently and effectively at the early stage.

Description

technical field [0001] The invention relates to the field of medical testing, in particular to a method for preparing a cTnI, NT-proBNP and D-dimer chest pain triple colloidal gold immune test strip. Background technique [0002] cTnI: cTn (cardiac troponin) exists in cardiomyocytes and consists of three subunits: cTnI (cardiac troponin I), cTnT (cardiac troponin T) and cTnC (cardiac troponin C). cTnI and cTnT are cardiomyocyte-specific antigens. After myocardial cell injury, cTnI and cTnT appeared in the blood at the earliest time and lasted for a long time. cTnI and cTnT have high sensitivity and specificity to myocardial injury, and cTnI is more sensitive than cTnT. Its molecular weight is 23000. After myocardial injury, 3 cTnI rises within ~4 hours, peaks at 11-24 hours, and returns to normal within 7-10 days. cTnI has now replaced other early myocardial markers as the preferred myocardial injury marker for the diagnosis of ACS (acute coronary syndrome), especially for...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/558
CPCG01N33/558G01N33/6803G01N2333/47
Inventor 刘丽王玉金杨书豪杜迅何蔚荭安明理李珊珊周伏忠胡宜亮陈国参孙晨阳
Owner HENAN ACAD OF SCI INST OF BIOLOGY LIABILITY
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