Kit for determining amino-terminal pro-brain natriuretic peptide, preparation method and detection method

A brain natriuretic peptide and amino-terminal technology, which is applied in the field of amino-terminal brain natriuretic peptide precursor assay kits, can solve problems such as low accuracy, human hazards, and poor reagent stability, so as to reduce batch-to-batch differences and maintain biological Activity, less impact on the environment

Pending Publication Date: 2020-11-24
深圳市爱康试剂有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are various problems in the current N-terminal BNP precursor detection methods: radioimmunoassay uses isotope labeling, which is very harmful to the human body and brings inconvenience to the experiment; magnetic particle chemiluminescence immunoassay, reagents The stability is poor, the detection precision is not high, and the instrument failure rate is high; the enzyme-linked immunoassay method, due to the instability of the enzyme itself, is greatly affected by other factors, and the repeatability is poor; the colloidal gold method has poor sensitivity and cannot Quantitative detection; fluorescent immunoassay, also has the problems of low accuracy and poor precision
This method is easy to cause the activity of the fluorescent conjugate coated on the unit area of ​​the conjugate gasket to be uneven, and the coefficient of variation CV of the reagent is relatively high.

Method used

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  • Kit for determining amino-terminal pro-brain natriuretic peptide, preparation method and detection method
  • Kit for determining amino-terminal pro-brain natriuretic peptide, preparation method and detection method
  • Kit for determining amino-terminal pro-brain natriuretic peptide, preparation method and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Preparation of reagent cards

[0074] (1) Preparation of detection line and quality control line: prepare amino-terminal brain natriuretic peptide precursor monoclonal antibody concentration of 2.0 mg / mL and goat anti-chicken IgY antibody concentration of 1 mg / mL respectively, and use a spray film machine to draw lines to On the test line and quality control line area on the nitrocellulose membrane, the coating volume is 1.0 μL / cm, put the scratched nitrocellulose membrane in a blast drying oven, and dry it at 50°C for 18 hours;

[0075] (2) Preparation of the sample pad: Prepare a blocking solution, add the blocking solution to the glass fiber to make the glass fiber completely wet, place it in a blast drying oven, and dry it at 50°C for 18 hours;

[0076] (3) Assembling of the reagent card: stick the nitrocellulose membrane to the backing plate, one end of the sample pad and one end of the water-absorbing pad are respectively lapped on the two ends of the nitrocellulo...

Embodiment 2

[0102] Preparation of reagent cards

[0103] (1) Preparation of test line and quality control line: prepare N-terminal BNP precursor monoclonal antibody concentration at 3.0 mg / mL and goat anti-rabbit IgG antibody concentration at 2 mg / mL respectively, and use a spray film machine to mark the line to On the test line and quality control line area on the nitrocellulose membrane, the coating volume is 0.8μL / cm, put the scratched nitrocellulose membrane in a blast drying oven, and dry it at 45°C for 20 hours;

[0104] (2) Preparation of the sample pad: Prepare a blocking solution, add the blocking solution to the glass fiber to make the glass fiber completely wet, place it in a blast drying oven, and dry it at 45°C for 20 hours;

[0105] (3) Assembling of the reagent card: stick the nitrocellulose membrane to the backing plate, one end of the sample pad and one end of the water-absorbing pad are respectively lapped on the two ends of the nitrocellulose membrane, cut into test str...

Embodiment 3

[0128] Preparation of reagent cards

[0129] (1) Preparation of test line and quality control line: Prepare N-terminal BNP precursor monoclonal antibody concentration at 0.5 mg / mL and goat anti-mouse IgG antibody concentration at 0.5 mg / mL respectively, and use a spray film machine to draw the line To the test line and quality control line area on the nitrocellulose membrane, the coating volume is 0.5 μL / cm, put the scratched nitrocellulose membrane in a blast drying oven, and dry it at 37°C for 24 hours;

[0130] (2) Preparation of the sample pad: Prepare a blocking solution, add the blocking solution to the glass fiber to make the glass fiber completely wet, place it in a blast drying oven, and dry it at 37°C for 24 hours;

[0131] (3) Assembling of the reagent card: stick the nitrocellulose membrane to the backing plate, one end of the sample pad and one end of the water-absorbing pad are respectively lapped on the two ends of the nitrocellulose membrane, cut into test stri...

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Abstract

The invention discloses a kit for determining an amino-terminal pro-brain natriuretic peptide. The kit is characterized by comprising a reagent card, a reaction substance and a reaction buffer solution, a detection line of the reagent card is coated with an amino-terminal pro-brain natriuretic peptide monoclonal antibody, and a quality control line of the reagent card is coated with one of a goatanti-chicken IgY antibody, a goat anti-rabbit IgG antibody or a goat anti-mouse IgG antibody. Before the kit is used, a reaction buffer solution is adopted to redissolve a reaction substance, and thena sample to be detected is added so that the amino-terminal pro-brain natriuretic peptide in the sample to be detected fully reacts with the amino-terminal pro-brain natriuretic peptide monoclonal antibody marked by the fluorescent microspheres, and the kit is high in detection accuracy, strong in specificity, good in reproducibility and rapid to operate.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis and detection, in particular to an amino-terminal brain natriuretic peptide precursor assay kit, a preparation method and a detection method. Background technique [0002] N-terminal pro-brain natriuretic peptide (NT-proBNP) is mainly released by ventricular cells in response to myocardial stress and filling pressure, and plays a role in balancing intravascular volume. After cardiomyocytes are stimulated, brain natriuretic peptide is released in the form of prohormone (proBNP), and then it is cleaved into amino-terminal brain natriuretic peptide precursor (NT-proBNP) containing 76 amino acids under the action of endonuclease and C-terminal polypeptide BNP containing 32 amino acids. BNP has an important biological activity because of a characteristic amino acid ring, but its half-life in vivo is short, only 18-22 minutes; while NT-proBNP has a linear structure, no biological activity, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/74G01N33/577G01N33/558G01N33/543G01N33/533
CPCG01N33/74G01N33/577G01N33/558G01N33/533G01N33/543G01N2333/58
Inventor 肖宇强马金玉张平赖鹏飞
Owner 深圳市爱康试剂有限公司
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