Amino-terminal pro-brain natriuretic peptide detection kit, and preparation method and application thereof

A detection kit, a technology of brain natriuretic peptide, applied in the field of biochemical detection, can solve the problems of low sensitivity and only

Pending Publication Date: 2020-12-01
SHANGHAI UPPER BIO TECH PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And the performance disclosed in CN102854321A, the sensitivity is not more than 15pg / ml, the linear upper limit is 20000pg / ml
The detection sensitivity of the kit disclosed in the CN107656071A document is still only 20pg / ml for NT-proBNP, and the reporting time needs 15min

Method used

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  • Amino-terminal pro-brain natriuretic peptide detection kit, and preparation method and application thereof
  • Amino-terminal pro-brain natriuretic peptide detection kit, and preparation method and application thereof
  • Amino-terminal pro-brain natriuretic peptide detection kit, and preparation method and application thereof

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preparation example Construction

[0063]

[0064] In another embodiment, the present invention provides a method for preparing an NT-proBNP detection kit.

[0065] In one embodiment, the method provided by the present invention comprises the steps of coating the first antibody strain on magnetic microspheres and labeling the second antibody strain with a tracer.

[0066] In another embodiment, the method provided by the present invention includes the following steps: magnetic microsphere suspension coated with NT-proBNP monoclonal antibody; preparing NT-proBNP monoclonal antibody solution labeled with acridinium ester; preparing chemiluminescence excitation solution; preparing Cleaning fluid.

[0067] In a preferred embodiment, the method provided by the present invention further includes the step of preparing a calibrator.

[0068]

[0069] In another embodiment, the present invention provides the use of a NT-proBNP detection kit.

[0070] In one embodiment, the present invention provides a first antibo...

Embodiment 1

[0075] Embodiment 1: the preparation method of cardiac muscle protein detection kit

[0076] Preparation 1: The first antibody-coated magnetic microspheres

[0077] (1) Measure 10mg of magnetic microspheres (average particle size 1.5μm, purchased from Bangs Laboratories, solid content 2.54%), and suspend in 1mL of 0.1M MES buffer, magnetize for 5-10min, discard After the supernatant, repeat the above washing steps 3-5 times, add 1ml of the above buffer (0.1M MES buffer), and vortex to mix.

[0078] (2) Add 200 μg of NT-proBNP first monoclonal antibody to make the mass ratio of magnetic microspheres to antibody 50:1, vortex and mix, and incubate at 37°C for 1 hour.

[0079] (3) Add 10 μL of 10 mg / ml 1-(3-dimethylaminopropyl)-3-ethyldiimine hydrochloride (EDC), vortex and incubate at 37° C. for 1.5 h.

[0080] (4) Take 200 μL of PBS solution (0.02M) containing 1% casein for blocking, and block for 2 hours.

[0081] (5) Add 1ml of luminescence recovery solution 1 (5g of bovi...

Embodiment 2

[0096] Embodiment 2: the use method of NT-proBNP detection kit

[0097] Use the first strain of antibody obtained in preparation 1 to coat magnetic microspheres, the second strain of antibody obtained in preparation 2 to label acridinium ester, the high and low point calibrator of preparation 3, the chemiluminescent substrate solution of preparation 4, and the magnetic The microsphere washing solution was used to prepare the detection kit of amino-terminal brain natriuretic peptide precursor.

[0098] After using the kit to detect the sample, the concentration of NT-proBNP in the sample was detected by the Yaoxuanfeng automatic chemiluminescence immunoassay analyzer.

[0099] Specifically, follow the steps below to measure the concentration of NT-proBNP in the sample to be tested using the Yao Xuanfeng automatic chemiluminescence immunoassay analyzer:

[0100] 1. Dilute the first antibody-coated magnetic microspheres prepared in Preparation 1 to 0.05 mg / ml according to the ...

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Abstract

The invention relates to an amino-terminal pro-brain natriuretic peptide direct chemiluminiscence detection kit and a preparation method and application thereof. Particular, the kit provided by the invention comprises a first antibody coating magnetic microspheres and a second antibody marked with a tracing marker, wherein the first antibody and the second antibody are monoclonal antibodies of anti-amino-terminal pro-brain natriuretic peptide, and the specific binding sites of the amino-terminal pro-brain natriuretic peptide recognized by the first antibody and the second antibody are different. The kit can be used for determining whether a to-be-detected sample contains the amino-terminal pro-brain natriuretic peptide and/or the content of the amino-terminal pro-brain natriuretic peptide,can be used for accurately diagnosing the severity of heart failure as soon as possible, provides sufficient time for heart failure treatment, and provides sufficient support for prognosis detectionof heart failure patients.

Description

technical field [0001] The invention relates to the field of biochemical detection, in particular to a direct chemiluminescent detection kit for amino terminal brain natriuretic peptide precursor, a preparation method and an application. Background technique [0002] Since BNP (brain natriuretic peptide) was isolated from pig brain in 1988, and NT-proBNP (amino-terminal brain natriuretic peptide precursor) was found in human plasma in 1995, researchers have found 6 natriuretic peptides in vertebrates. peptide subtype. When ventricular volume increases, cardiac pressure load increases, myocardial ischemia, and ventricular wall muscle tension increase, the BNP gene in cardiomyocytes is activated to generate BNP precursor (proBNP). When the precursor enters the blood, it is cleaved into NT-proBNP and bioactive hormone BNP under the action of protease. The detection of NT-proBNP is not affected by the position, time and daily activities, and the half-life is as long as 60-120m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/536G01N21/76
CPCG01N33/6893G01N33/6887G01N33/577G01N33/536G01N21/76G01N2800/32
Inventor 张瑞石晓强徐建新李福刚周雪
Owner SHANGHAI UPPER BIO TECH PHARMA
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