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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 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

Method used

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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

Embodiment 1

[0078] Embodiment 1: Preparation of NGAL detection kit

[0079] This application exemplarily prepared three kinds of NGAL detection kits, and its preparation raw 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] R2 preparation process is as follows:

[0090] Small particle size latex reagent preparation:

[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 antibody concentration of 2mg / ml;

[0092] 2. Dilute 4ml of 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 prepar...

Embodiment 2

[0188] Example 2: NGAL detection kit measurement steps, drawing of the calibration curve, comparative research with a well-known brand product on the market, the manufacturer’s reagent detection principle is also latex enhanced immune turbidimetry, the main components: 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 assay steps of the present application are shown in Table 1.

[0190] Table 1: Determination steps of the application kit:

[0191]

[0192] Taking the kit 2 of Example 1-(2) as an example, the standard concentration is the horizontal axis, and the corresponding ΔOD 570 As the vertical axis, use nonlinear fitting, such as spline to draw the standard curve, the result is as follows figure 1 shown.

Embodiment 3

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

[0194]Linearity test: using methods known to those skilled in the art, a certain high NGAL concentration sample (urine sample at 2 hours after surgery for acute kidney injury patients) was doubly diluted, using the method shown in Example 1-(2) Kit 2 is a detection kit, which measures the concentration after dilution, and calculates the average value for three times, and compares it with the theoretical concentration to calculate its linear deviation. At the same time, compared with a well-known brand in the above market, the measurement results are shown in Table 2, Table 3, Table 4 and Table 5.

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

[0196]

[0197] Table 3: High range linearity of control reagents: unit ng / ml

[0198] Dilution factor

measurement 1

measurement 1

measurement 1

Measure mean

theoretical value

Linear devia...

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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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