Method, reagent and kit for quantitative determination of NGAL content in human serum

A technology for lipocalin and neutrophils, which is applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of high price, unsuitable for routine inspection, waste of resources, etc., and achieves easy operation and fully automated analysis. Effect

Inactive Publication Date: 2015-08-26
ZHEJIANG LANSEN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Currently, only enzyme immunoassay (ELISA) is the only clinically used detection method for NGAL. This method has the disadvantages of cumbersome operation, sample pretreatment, long detection time, high price, and excessive waste of resources. It is not suitable for routine testing, especially on a large scale. Simultaneous detection of many items in epidemiological examination or clinical mass specimens

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  • Method, reagent and kit for quantitative determination of NGAL content in human serum
  • Method, reagent and kit for quantitative determination of NGAL content in human serum

Examples

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Effect test

Embodiment 1

[0023] Configure the following reagent I and reagent II of the present invention according to the following ingredients and ratios:

[0024] Reagent I:

[0025] Phosphate buffer 30mmol / L

[0026] Polyethylene glycol 8000 30mmol / L

[0027] Reagent II:

[0028] Mouse anti-human NGAL monoclonal antibody latex particles 50mL / mol

[0029] Antibody stabilizer 0.05g / mL

[0030] Mix 240 μl reagent I and 3 μl serum sample in a sample tube, incubate at 37°C for 5 minutes, use Hitachi 7060 automatic biochemical analyzer, measure the absorbance A1 of the sample relative to the blank tube at a wavelength of 546 nm, and then add Mix 60 μl of reagent II, incubate at 37°C for 5 minutes, and measure the absorbance A2 of the sample relative to the blank tube. Calculate the ΔA sample according to the following formula (1). Use the same method and conditions to measure the absorbance value of the standard solution, and calculate the △A standard. Then calculate the neutrophil gelatinase-ass...

Embodiment 2

[0035] Configure the following reagent I and reagent II of the present invention according to the following ingredients and ratios:

[0036] Reagent I:

[0037] Phosphate buffer 150mmol / L

[0038] Polyethylene glycol 8000 150mmol / L

[0039] Reagent II:

[0040] Mouse anti-human NGAL monoclonal antibody latex particles 500mL / mol

[0041] Antibody stabilizer 0.5g / mL

[0042] Mix 240 μl reagent I and 3 μl serum sample in the sample tube, incubate at 37°C for 5 minutes, use the Olympus 400 automatic biochemical analyzer, measure the absorbance A1 of the sample relative to the blank tube at a wavelength of 546 nm, and then Add 60 μl of reagent II to the sample, mix well, incubate at 37°C for 5 minutes, and measure the absorbance A2 of the sample relative to the blank tube. Calculate the ΔA sample according to the following formula (1). Use the same method and conditions to measure the absorbance value of the standard solution, and calculate the △A standard. Then calculate th...

Embodiment 3

[0047] Configure the following reagent I and reagent II of the present invention according to the following ingredients and ratios:

[0048] Reagent I:

[0049] Phosphate buffer 100mmol / L

[0050] Polyethylene glycol 8000 50mmol / L

[0051] Reagent II:

[0052] Mouse anti-human NGAL monoclonal antibody latex particles 300mL / mol

[0053] Antibody stabilizer 0.1g / mL

[0054] Mix 240 μl reagent I and 3 μl serum sample in the sample tube, incubate at 37°C for 5 minutes, use Beckman LX20 automatic biochemical analyzer, measure the absorbance A1 of the sample relative to the blank tube at a wavelength of 546 nm, and then add to the sample Add 60 μl of reagent II, mix well, incubate at 37°C for 5 minutes, and measure the absorbance A2 of the sample relative to the blank tube. Calculate the ΔA sample according to the following formula (1). Use the same method and conditions to measure the absorbance value of the standard solution, and calculate the △A standard. Then calculate th...

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Abstract

The invention relates to a reagent for the quantitative determination of NGAL (Neutrophil Gelatinase-Associated Lipocalin) content in human serum. The reagent comprises a reagent I and a reagent II which are placed separately, wherein the reagent I contains T phosphate buffer and polyethylene glycol-6000; the reagent II contains mouse anti-human NGAL monoclonal antibody latex particles and an antibody stabilizer. A kit and a detection method adopted in the invention only require a few microlitres of serum, need no centrifugation or electrophoresis and other separation treatments, are easy to operate, can meet the requirements of automatic analysis, and are applicable to the timely and accurate detection of large-scale samples.

Description

technical field [0001] The application relates to a quantitative determination method, reagent and kit for the content of neutrophil gelatinase-associated lipocalin in human serum. Background technique [0002] Neutrophil gelatinase-associated lipocalin rapid detection ELISA kit can be used for human urine, plasma and serum detection. Acute renal failure (ARF) is a common complication of cardiac surgery, nephrotoxicity, and kidney transplantation. [0003] Neutrophil gelatinase-associated lipocalin (neutrophil gelatinase-associated lipocalin, NGAL), also known as Lipocalin-2 (lipocalin-2), is a new member of the lipocalin family, is the main cause of renal dysfunction Early biomarkers. NGAL not only exists in neutrophils, but also in specific epithelial cells. For example, in the process of ischemic and toxic renal injury, NGAL in renal tubular epithelial cells will increase significantly. Within hours, the levels of NGAL in urine and blood will increase significantly, so...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6893
Inventor 张益军
Owner ZHEJIANG LANSEN BIOTECH
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