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Neutrophil gelatinase-associated lipocalin determination kit

A lipocalin and neutrophil technology, which is applied in the fields of medicine and biochemistry, can solve the problems of inconvenience, difficulty in meeting detection requirements, and inability to meet wide linear range and high sensitivity at the same time, so as to improve the reaction rate, increase the The effect of improving usability and stability

Active Publication Date: 2018-04-17
浙江夸克生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are kits for the determination of NGAL concentration in human plasma or urine by latex-enhanced immunoturbidimetry, but these kits cannot meet the requirements of wide linear range and high sensitivity at the same time, so it is necessary to dilute the sample to measure high concentration of NGAL , which is inconvenient to use
Existing latex-enhanced immunoturbidimetric method, such as the detection sensitivity of the kits disclosed in patent CN 104198732 and patent CN 102680698, is greater than or equal to 10ng / ml, and the linear range is below 6000ng / ml, which are difficult to meet the clinical detection needs

Method used

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  • Neutrophil gelatinase-associated lipocalin determination kit
  • Neutrophil gelatinase-associated lipocalin determination kit
  • Neutrophil gelatinase-associated lipocalin determination kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1 kit preparation 1

[0040] Reagent R1: pH7.4

[0041] Tris 100mM

[0042] Tween-20 1%

[0043] Polyethylene glycol 6000 4g / l

[0044] Sodium chloride 15g / l

[0045] The preparation process of reagent R2 is as follows:

[0046] (1) Dilute polystyrene particles with carboxyl functional groups on the surface and 0.8 μm in diameter with MES buffer to a final concentration of 0.5-3% (wt), add 50-90mM EDAC, mix well, and shake at room temperature for 30 -80 minutes;

[0047] (2) Centrifuge the mixture obtained in step (1), discard the supernatant, wash twice with MES buffer, and finally resuspend the latex particles in MES buffer to a final concentration of 2%, and disperse them by ultrasonic to obtain a particle dispersion ;

[0048] (3) Dilute the NGAL antibody with MES buffer, mix it with the particle dispersion in step (2) in equal volume, and react at room temperature for 2-4 hours;

[0049] (4) After centrifuging the reaction solution in step (3) at...

Embodiment 2

[0063] Embodiment 2 kit preparation 2

[0064] Reagent R1: pH7.4

[0065] Tris 100mM

[0066] Tween-20 1%

[0067] Polyethylene glycol 6000 4g / l

[0068] Sodium chloride 15g / l

[0069] The preparation process of reagent R2 is as follows:

[0070] (1) Dilute polystyrene particles with carboxyl functional groups on the surface and 1.0 μm in diameter with MES buffer to a final concentration of 0.5-3% (wt), add 50-90mM EDAC, mix well, and shake at room temperature for 30 -80 minutes;

[0071] (2) Centrifuge the mixture obtained in step (1), discard the supernatant, wash twice with MES buffer, and finally resuspend the latex particles in MES buffer to a final concentration of 2%, and disperse them by ultrasonic to obtain a particle dispersion ;

[0072] (3) Dilute the NGAL antibody with MES buffer, mix it with the particle dispersion in step (2) in equal volume, and react at room temperature for 2-4 hours;

[0073] (4) After centrifuging the reaction solution in step (3) at...

Embodiment 3

[0089] Embodiment 3 kit preparation 3

[0090] Reagent R1: pH7.4

[0091] Tris 100mM

[0092] Tween-20 1%

[0093] Polyethylene glycol 6000 4g / l

[0094] Sodium chloride 15g / l

[0095] The preparation process of reagent R2 is as follows:

[0096] (1) Dilute polystyrene particles with carboxyl functional groups on the surface and 1.0 μm in diameter with MES buffer to a final concentration of 0.5-3% (wt), add 50-90mM EDAC, mix well, and shake at room temperature for 30 -80 minutes;

[0097] (2) Centrifuge the mixture obtained in step (1), discard the supernatant, wash twice with MES buffer, and finally resuspend the latex particles in MES buffer to a final concentration of 2%, and disperse them by ultrasonic to obtain a particle dispersion ;

[0098] (3) Dilute the NGAL antibody with MES buffer, mix it with the particle dispersion in step (2) in equal volume, and react at room temperature for 2-4 hours;

[0099] (4) After centrifuging the reaction solution in step (3) at...

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Abstract

The invention belongs to the technical field of medicine and biochemistry and in particular relates to a neutrophil gelatinase-associated lipocalin determination kit. The kit is composed of a reagentR1 and a reagent R2, wherein the reagent R1 contains biological buffer, a coagulation accelerator, a surfactant, an osmotic pressure regulator and water; and the reagent R2 contains latex particles coating a neutrophil gelatinase-associated lipocalin antibody, biological buffer, a sealing agent, a preservative and water. The neutrophil gelatinase-associated lipocalin detection kit performs determination by adopting particle-enhanced turbidimetric immunoassay, and compared with the traditional neutrophil gelatinase-associated lipocalin (NGAL) detection kit, the detection kit disclosed by the invention has the advantages of being rapid, sensitive, excellent in accuracy, high in specificity, excellent in stability and the like.

Description

technical field [0001] The invention relates to the technical fields of medicine and biochemistry, in particular to a kit for measuring neutrophil gelatinase-associated lipocalin. Background technique [0002] In recent years, with the improvement of people's living standards and changes in living and eating habits, the incidence of kidney disease has increased year by year. Most kidney diseases have complex etiology, protracted disease course, and increase the risk of cardiovascular disease, which has become a global public health problem. Kidney disease can be divided into two main syndromes - acute kidney injury (acute kidney injury, AKI) and chronic kidney disease (chronic kidney disease, CKD). AKI refers to a group of clinical syndromes of sudden and continuous decline in renal function, manifested as azotemia, water-electrolyte and acid-base balance disorders, and symptoms of various systems throughout the body, which may be accompanied by oliguria or anuria, and is a...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/54313G01N33/6893G01N2800/347
Inventor 吴正伟章晓庆
Owner 浙江夸克生物科技有限公司
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