Assay kit for neutrophil gelatinase-associated lipocalin

A detection kit and kit technology, applied in the field of neutrophil gelatinase-associated lipocalin detection kits, can solve problems such as inability to satisfy wide linear range and high sensitivity at the same time, inconvenient use, etc., and achieve improved detection limit , improve sensitivity, and expand the effect of linear range

Active Publication Date: 2012-07-18
BEIJING STRONG BIOTECH
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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 requirem

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  • Assay kit for neutrophil gelatinase-associated lipocalin
  • Assay kit for neutrophil gelatinase-associated lipocalin
  • Assay kit for neutrophil gelatinase-associated lipocalin

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0078] Example 1: Preparation of NGAL detection kit

[0079] This application exemplarily prepares three NGAL detection kits, the preparation materials, preparation steps, and performance are as follows:

[0080] (1) Kit 1:

[0081] R1: pH 7.3,

[0082] The main ingredients are:

[0083] Glycine 50mM,

[0084] NaCl 0.15M,

[0085] NaN3 0.1%,

[0086] EDTA 10mM,

[0087] BSA 0.1%,

[0088] Rabbit IgG 1%,

[0089] The preparation process of R2 is as follows:

[0090] Preparation of small particle size latex reagent:

[0091] 1. Dilute 4ml of rabbit anti-human NGAL polyclonal antibody (10mg / ml) with 20mM HEPES solution (pH 7.0) at room temperature to a final concentration of 2mg / ml;

[0092] 2. Dilute 4ml 74nm latex solution (10%wt) with 20mM HEPES solution (pH 7.0) at room temperature to make the latex concentration 4%;

[0093] 3. Mix the diluted antibody and latex, shake at room temperature for 30 minutes, add 10ml of 5mg / ml EDAC solution prepared with 20mM HEPES solution (pH 7.0), react at room ...

Example Embodiment

[0188] Example 2: NGAL detection kit determination steps, calibration curve drawing, comparison study with a well-known brand product on the market, the manufacturer's reagent detection principle is also the latex enhanced immunoturbidimetric method, the main composition: R1, 10mmol Tris , 0.1% NAN3, 0.5% BSA; R2, 0.15% latex suspension coated with anti-human NGAL monoclonal antibody, the comparison is as follows:

[0189] The measurement steps of this application are shown in Table 1.

[0190] Table 1: The determination steps of the kit of this application:

[0191]

[0192] Take the kit 2 of Example 1-(2) as an example, the concentration of the standard is on the horizontal axis, and the corresponding ΔOD 570 Is the vertical axis, using nonlinear fitting, such as spline to draw a standard curve, the result is as figure 1 Shown.

Example Embodiment

[0193] Example 3: Linearity and minimum detection limit of NGAL kit

[0194] Linearity experiment: using a method known to those skilled in the art, a sample with a high NGAL concentration (a urine sample of a patient with acute kidney injury after surgery at 2 hours) is diluted by multiples, as shown in Example 1-(2) Reagent kit 2 is a detection kit. Determine the concentration after dilution, determine the average value for three times, compare with the theoretical concentration, and calculate the linear deviation. At the same time, it was compared with a well-known brand in the above-mentioned market, and the measurement results are shown in Table 2, Table 3, Table 4 and Table 5.

[0195] Table 2 Linearity of high value range of kit 2 of this application: unit ng / ml

[0196]

[0197] Table 3: Linearity of the high value range of the control reagent: unit ng / ml

[0198] Dilution factor

[0199] Table 4: Linearity of low value range of kit 2 of this application: unit ng / ml

[0200...

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Abstract

The invention relates to an assay kit for neutrophil gelatinase-associated lipocalin (NGAL), in particular to an assay kit for detecting the concentration of the NGAL in human body fluids and a preparation method for the assay kit. The kit comprises a reagent R1, a reagent R2 and a working calibrator, wherein the reagent R1 is a buffer solution which is suitable for antigen-antibody reaction; and the reagent R2 is a solution simultaneously containing nanospheres which have two particle sizes and are combined with an anti-human NGAL antibody, and ensures that the nanospheres are coagulated to cause the change of the turbidity through antigen-antibody specific reaction, and the NGAL can be quantificationally determined by contrasting calibration curves. The kit has relatively high sensitivity when determining low NGAL concentration and has a relatively wide linear range when determining high NGAL concentration, so a prozone phenomenon is avoided. The kit has stable physical and chemical properties, is convenient to use, and has a broad application prospect in clinical laboratory science.

Description

technical field [0001] The application relates to a neutrophil gelatinase-associated lipocalin assay kit, more specifically, the application relates to a method for detecting neutrophil gelatinase-associated lipocalin (NGAL) in human body fluid A detection reagent for concentration and its preparation method, the reagent includes reagent R1, reagent R2 and working calibrator, reagent R1 is a buffer solution suitable for antigen-antibody reaction, R2 is a combination of anti-human NGAL containing two particle sizes Antibody nano-microsphere solution, the reagent causes the nano-microspheres to aggregate through the specific reaction of the antigen and antibody, resulting in a change in turbidity, and the calibration curve can be used for quantitative determination of NGAL. Background technique [0002] Neutrophil gelatinase-associated lipocalin (NGAL), also known as lipocalin-2 (Lipocalin-2), is a new member of the lipocalin family, which is the Early biomarkers of functiona...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/53G01N33/52G01N33/54346
Inventor 张小锐蔡华雅刘希
Owner BEIJING STRONG BIOTECH
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