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Kit for detecting brain natriuretic peptide by nano microsphere immunonephelometry

A technology of nano-microspheres and immune turbidimetry, which is applied in biological testing, measuring devices, material inspection products, etc. It can solve the problems of sample pretreatment, radiation radiation, and high cost, and achieve good accuracy and strong anti-interference ability , the effect of simple operation

Active Publication Date: 2010-09-01
浙江伊利康生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Known methods for measuring BNP include radioimmunoassay (IRA), immunoradiometry (IRMA), electrochemiluminescence (ECLA), enzyme-linked immunoassay (ELISA); radioimmunoassay directly without extracting plasma BNP Measurement, this assay system uses two anti-human BNP monoclonal antibodies, one recognizes the C-terminal sequence of BNP, and the other recognizes its ring structure, that is, the plasma BNP concentration is determined by the sandwich method. Although this method is more sensitive, accurate and easy operation; but this method has the same problems as the immunoradiometric method, such as radiation exposure and pollution
Electrochemiluminescence method is more sensitive, accurate and precise than radioimmunoassay, but it is expensive and requires a matching chemiluminescence instrument for detection
Enzyme immunoassay (ELISA) has disadvantages such as cumbersome operation and sample pretreatment

Method used

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  • Kit for detecting brain natriuretic peptide by nano microsphere immunonephelometry
  • Kit for detecting brain natriuretic peptide by nano microsphere immunonephelometry
  • Kit for detecting brain natriuretic peptide by nano microsphere immunonephelometry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. BNP solution (reagent R 1 )

[0030] Tris Buffer 50mmol / L

[0031] Tween-20 (surfactant) 0.5mmol / L

[0032] NaCl (electrolyte) 100mmol / L

[0033] PEG-6000 (reaction accelerator) 4mmol / L

[0034] Broad-spectrum fungicide (preservative) 3mmol / L

[0035] Disodium EDTA (stabilizer) 5mmol / L

[0036] The rest is purified water

[0037] 2. Anti-human BNP antibody solution (reagent R 2 )

[0038] Use 100mmol / L tris buffer (pH 7.4) to dilute the goat anti-human BNP antibody polyclonal antibody and 80nm nanospheres (the weight ratio of antibody and microspheres is 1:1) at room temperature, and mix the two After mixing, shake and adsorb at room temperature for 2 hours (that is, physical adsorption method known to those skilled in the art), add blocking solution (containing 0.1% BSA trishydroxymethylaminotetraxane buffer) to block for 1 hour, centrifuge to remove the supernatant, and use Dilute the 100mmol / L tris buffer solution to a concentration of 0.3%, and add an ap...

Embodiment 2

[0049] 1. BNP solution (reagent R 1 )

[0050] Tris Buffer 100mmol / L

[0051] Tween-20 (surfactant) 4mmol / L

[0052] NaCl (electrolyte) 60mmol / L

[0053] PEG-6000 (reaction accelerator) 8mmol / L

[0054] Broad-spectrum fungicide (preservative) 0.1mmol / L

[0055] Disodium EDTA (stabilizer) 0.5mmol / L

[0056] The rest is purified water

[0057] 2. Anti-human BNP antibody solution (reagent R 2 )

[0058] Use 150mmol / L Tris buffer (pH 7.4) to dilute goat anti-human BNP antibody polyclonal antibody and 120nm nanometer microspheres (the weight ratio of antibody and microsphere is 1: 1) at room temperature, mix the two After mixing, shake and adsorb at room temperature for 2 hours (that is, physical adsorption method known to those skilled in the art), add blocking solution (containing 0.1% BSA trishydroxymethylaminotetraxane buffer) to block for 1 hour, centrifuge to remove the supernatant, and use Dilute the 100mmol / L tris buffer solution to a concentration of 4%, and add a...

Embodiment 3

[0069] 1. BNP solution (reagent R 1 )

[0070] Tris (buffer) 200mmol / L

[0071] Tween-20 (surfactant) 2mmol / L

[0072] NaCl (electrolyte) 200mmol / L

[0073] PEG-6000 (reaction accelerator) 5mmol / L

[0074] Broad-spectrum fungicide (preservative) 3mmol / L

[0075] Disodium EDTA (stabilizer) 2mmol / L

[0076] The rest is purified water

[0077] 2. Anti-human BNP antibody solution (reagent R 2 )

[0078] Use 200mmol / L Tris buffer (pH 7.4) to dilute the goat anti-human BNP antibody polyclonal antibody and 100nm nanospheres at room temperature (the weight ratio of antibody and microspheres is 1: 1), and mix the two After mixing, shake and adsorb at room temperature for 2 hours (that is, physical adsorption method known to those skilled in the art), add blocking solution (containing 0.1% BSA trishydroxymethylaminotetraxane buffer) to block for 1 hour, centrifuge to remove the supernatant, and use 100mmol / L tris buffer solution is diluted to concentration and is 2%, and the an...

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Abstract

The invention relates to a kit for detecting brain natriuretic peptide in blood serum and provides the kit for detecting brain natriuretic peptide by nano microsphere immunonephelometry, which has the characteristics of no need of dilution of a sample, simple operation, high precision, good repeatability and suitability for a clinically common used automatic biochemical analyzer, in order to solve technical problems. The invention adopts a technical scheme that the kit for detecting the brain natriuretic peptide by nano microsphere immunonephelometry comprises the following components: a, a reagent R1 which comprises buffer solution, a preservative, a stabilizing agent, an electrolyte, a surfactant, and the balance of water; b, a reagent R2 which comprises buffer solution, nano microspheres combined with brain natriuretic peptide antibodies and a preservative, wherein the diameter of the microspheres is 50 to 150 nm; and c, a reference calibration material which comprises buffer solution, a stabilizing agent, a preservative, a recombinant brain natriuretic peptide pure product which is added in a corresponding amount according to the required concentration of the BNP (brain natriuretic peptide) reference calibration material, and the balance of water.

Description

technical field [0001] The invention relates to a test kit for measuring serum components, in particular to a test kit for measuring brain natriuretic peptide (BNP) in serum, which can be widely used in the technical fields of medicine and biochemistry. Background technique [0002] Brain Natriuretic Peptide (BNP), also known as B-type Natriuretic Peptide, is one of the structurally related peptide hormone family natriuretic peptides produced by heart cells. It consists of 32 amino acids, including a ring structure composed of 17 amino acids through a pair of disulfide bonds. BNP is widely distributed in brain, spinal cord, heart and lung and other tissues, among which the heart has the highest content. The content of BNP in the medulla oblongata is the highest in the brain, and the BNP in the heart mainly exists in the left and right atrium. A small amount of BNP is also contained in the walls of arteries, hepatic arteries and pulmonary veins. When the heart is insuffici...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/544G01N33/531
Inventor 王贤理蒙凯蔡其浩
Owner 浙江伊利康生物技术有限公司
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