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ProANP (pro-atrial natriuretic peptide) rapid test paper and related application

A technology for testing test strips and test strip cards, which is applied in the field of clinical testing and can solve the problems of poor quantitative accuracy, inability to quantify, and poor repeatability of ELISA methods

Inactive Publication Date: 2017-05-17
GUANGZHHOU HUAHONG BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the detection methods for proANP are mainly enzyme-linked immunoassay (ELISA) and radioimmunoassay; and mainly rely on imports, but ELISA has poor quantitative accuracy, long operation time, and low degree of automation, and is mostly used for qualitative detection; radioimmunoassay The analytical method has high sensitivity, simple operation, and accurate measurement. The disadvantage is that it takes a long time, and there is a risk of radioactive pollution and radiation. Although the colloidal gold immunochromatography method has the advantages of less sample consumption, simplicity, speed, and low cost, its sensitivity is low. Low, generally only qualitative, not quantitative, especially the shortcoming of poor repeatability limits its clinical application, especially not suitable for the quantitative detection of body fluid marker proteins that need accurate quantification to help diagnose diseases

Method used

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  • ProANP (pro-atrial natriuretic peptide) rapid test paper and related application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Preparation of proANP detection test paper card

[0026] Preparation of nitrocellulose membrane coated with detection line and quality control line:

[0027] Use the coating diluent to adjust the concentration of proANP monoclonal antibody and goat anti-mouse IgG antibody to 5mg / ml, and the membrane liquid volume is 2μl / cm, and spray them in parallel on the nitrocellulose membrane as the detection line and quality control line respectively Coating is carried out, and the distance between the detection line and the quality control line is 5mm, and then placed in an oven, and dried at 37°C for 4 hours. The coating diluent was 100 mM Tris-HCl buffer containing 0.5% sodium lignosulfonate, pH 7.0.

[0028] Preparation of conjugation pads for absorbing antibodies labeled with fluorescent microspheres:

[0029] 1. Aldehydization of rare earth fluorescent microspheres:

[0030] Take 15mg of rare earth fluorescent microspheres, wash with 50mM, pH9.0 carbonate buffe...

Embodiment 2

[0037] Embodiment 2 test paper card linear range, precision and sensitivity test

[0038] Take the proANP standard solution with the content of 0.01, 0.05, 0.1, 0.5, 1, 5, 10 and 20nmol / L, and measure it with a test paper card.

[0039] Detection method: equilibrate the detection reagent and sample to room temperature, take out the test paper card (prepared in Example 1), and lay it flat; accurately draw 100 μl of standard sample, add it to the sample hole of the test paper card, and analyze it with fluorescent immunochromatography after 20 minutes instrument for testing.

[0040] Linearity: Establish a standard curve with fluorescence intensity values ​​and corresponding standard substance concentrations, specifically Y (fluorescence intensity)=116.27X+3.4, R2 value>0.999, indicating that the test paper card of the present invention has good linearity in the range of 0.01-2020nmol / L.

[0041] Sensitivity: PBS buffer was used as the blank control, and the fluorescence intensi...

Embodiment 3

[0043] Embodiment 3 stability test

[0044] Place the test paper card prepared in Example 1 in a 37°C environment for 3 months, then determine the standard curve and R of the kit according to the method in Example 2. 2 Value, and adopt the proANP standard substance of 1nmol / L to carry out precision investigation to kit (parallel determination 10 times, calculate CV%), specific result is as follows:

[0045]

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Abstract

The invention relates to ProANP (pro-atrial natriuretic peptide) rapid test paper and related application and belongs to the field of clinical test. The test paper is formed by: mutually overlapping a treated sample pad, a bonding pad which a rare-earth fluorescent microsphere labelled antibody is adsorbed to, a nitrocellulose film coated with a test line and quality control line and an absorbent pad sequentially on a PVC (polyvinyl chloride) board, and assembling.

Description

technical field [0001] The invention relates to a proANP rapid detection test paper card and related applications, belonging to the field of clinical detection. Background technique [0002] Heart failure is a group of syndromes caused by various structural or functional diseases of the heart that lead to impaired ventricular filling and / or ejection ability, and it is one of the most common causes of hospitalization or death in the elderly. With the aging of the population and the increase in the survival rate of myocardial infarction, heart failure is the only cardiovascular disease whose incidence and prevalence are still rising, and its prevention and treatment has become a hot spot in clinical cardiology research in recent years. [0003] Pro-atrial natriuretic peptide (proANP) is synthesized by cardiomyocytes and stored in secretory granules, and the main stimulus for hormone release into the blood circulation comes from the stretching of cardiomyocytes. When the atria...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/558G01N33/577
CPCG01N33/582G01N33/558G01N33/577
Inventor 陈立国邹伟权张亚丽李庆祥母润红王涛苏焱
Owner GUANGZHHOU HUAHONG BIOLOGICAL TECH
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