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Method for detecting neutrophil gelatinase-associated lipocalin (NGAL) in sample

A technology of lipocalin and neutrophils, which can be used in the measurement of color/spectral properties, measurement devices, biological tests, etc., and can solve the problems of detection within a day or even a few days after injury

Inactive Publication Date: 2011-05-25
WUHAN LIFE ORIGIN BIOTECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Common diagnostic methods such as measuring serum creatinine or cystatin C (Cystatin C) can only be detected within a day or even a few days after the injury

Method used

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  • Method for detecting neutrophil gelatinase-associated lipocalin (NGAL) in sample
  • Method for detecting neutrophil gelatinase-associated lipocalin (NGAL) in sample
  • Method for detecting neutrophil gelatinase-associated lipocalin (NGAL) in sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Detection of NGAL content in human urine by latex-enhanced immunoturbidimetric method

[0018] Realize the technical scheme of this example: an immunoturbidimetric method for detecting human NGAL, using anti-NGAL polyclonal antibody to firstly cross-link latex, and then use the cross-linked latex solution to react with NGAL of the specimen to be tested, the formed The complex causes a change in turbidity; the change is characterized by light projection intensity or light scattering intensity; the NGAL content of the sample to be tested is found from the standard curve of the concentration of the NGAL standard substance and the corresponding light scattering intensity or light projection intensity; the used The titer of polyclonal antibody to NGAL is at least 1:500, does not contain mixed antibodies, and can recognize multiple epitopes of NGAL; the specimen to be tested should be diluted 1:200-1:1000 with diluent;

[0019] In the above-mentioned method, describ...

Embodiment 2

[0020] Example 2 Determination of Human Urine NGAL Concentration by Sandwich ELISA

[0021] The specific steps are: coating NGAL antibody A; preparing a series of NGAL standard products; preparing human urine; adding a series of standard products or human urine with undetermined values ​​into the wells of a 96-microwell plate, and then making the labeled and binding sites different Bind to Antibody B coated on Antibody A on a 96-microwell plate, develop color with matrix solution for 20-30 minutes, stop color development, and measure the NGAL in the above series of standards and the human urine to be tested after 10 minutes. Absorbance; establish a standard curve of the concentration and absorbance of a series of standard substances and find out the concentration of NGAL in human urine to be determined from the curve.

Embodiment 3

[0022] Example 3 Determination of Human Urine NGAL Concentration by Competitive ELISA

[0023] The specific steps are: coating NGAL polyclonal antibody; preparing a series of NGAL standards; preparing human urine; mixing the series of standards or human urine with undetermined values ​​and labeled NGAL standards; developing color with matrix solution for 20-30 Stop the color development after 10 minutes, measure the absorbance of the above series of standard products and the NGAL reaction in the human urine to be detected after 10 minutes; establish a standard curve of the concentration and absorbance of the series of standard products and find out the human urine of the undetermined value from the curve The concentration of NGAL in the solution.

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Abstract

The invention relates to a method for detecting neutrophil gelatinase-associated lipocalin (NGAL) in a sample. The method comprises the following steps of: reacting an antibody of the NGAL with human NGAL of a specimen to be detected to form a compound which causes turbidity change; characterizing the change by using light projection intensity or light scattering intensity; and finding out the content of the human NGAL in the specimen to be detected from the concentration of the NGAL standard product and a standard curve of corresponding light scattering intensity or light transmission intensity. The method has the advantages of high sensitivity, high recovery rate and good repeatability, and a detection measure having the advantages of simplicity, convenience, good repeatability, strong adaptability and high speed is determined for measuring the NGAL in the samples of urine or blood plasma, and the like.

Description

technical field [0001] The invention relates to a method for detecting neutrophil gelatinase-associated lipocalin in a sample. Background technique [0002] Neutrophil gelatinase-associated lipocalin (NGAL for short) was first discovered in neutrophils in 1993, and is involved in inflammation, embryonic development, immune response, chemotaxis, signal transduction and various tumors Occurrence is related to processes such as development. In recent years, studies at home and abroad have shown that NGAL protein has the characteristics of specific expression changes in various disease processes, making NGAL a biomarker for detecting diseases. [0003] In the process of ischemic and toxic kidney injury, NGAL in the renal tubular epithelial cells will increase significantly, and within the first two hours, the level of NGAL in urine and blood will increase significantly, so NGAL is the early stage of acute kidney injury. sensitive markers. [0004] The prognosis of acute renal...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/539G01N33/577G01N33/546G01N21/31
Inventor 华权高沈鹤霄
Owner WUHAN LIFE ORIGIN BIOTECH LTD
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