Latex enhanced immunoturbidimetry NGAL detection kit

A detection kit and immunoturbidimetric technology, which is applied in the field of latex-enhanced immunoturbidimetric NGAL detection kits, can solve the problems of high reagent background detection linear range, poor reagent stability, and poor anti-interference ability, and achieve The effect of maintaining uniformity, maintaining stability, and wide linear range

Inactive Publication Date: 2014-12-17
上海睿康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the latex immunoturbidimetric method has great research value, but the latex-enhanced immunoturbidimetric assay kits currently on the market have poor anti-interference ability, low specificity, poor reagent stability, too high reagent background or detection Problems such as narrow linear range need to be improved

Method used

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  • Latex enhanced immunoturbidimetry NGAL detection kit
  • Latex enhanced immunoturbidimetry NGAL detection kit
  • Latex enhanced immunoturbidimetry NGAL detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] This embodiment relates to a NGAL detection kit and its preparation:

[0035] 1. Reagent R1:

[0036] pH7.0~7.6, 50~400mM buffer

MES buffer

pH7.4, 150mmol / L

Surfactant

Tween-20

50ml / L

Chelating agent

Disodium edetate

50mmol / L

Accelerator

Dextran 6000

200mmol / L

preservatives

NaN 3

1‰

[0037] Various ingredients can be added sequentially at room temperature, sealed and stored for later use. Alternatively, they are packaged individually and prepared immediately prior to testing.

[0038] 2. Reagent R2 is prepared as follows:

[0039] Step 1: The hydroxyl latex with a particle size of 80 nm was mixed with the hydroxylated latex of 160 nm, and activated with p-toluenesulfonyl cyanogen bromide chloride (50 mg / ml) in MOPSO (40 mM) solution at pH 6.0.

[0040] Step 2: Wash to remove the p-toluenesulfonyl chloride cyanogen bromide in the above solution, add pH 6.0 MOPSO (40mM) buffer, add ...

Embodiment 2

[0048] The NGAL detection kit described in this example is applicable to various types of automatic biochemical analyzers. Taking Hitachi 7170 automatic biochemical analyzer as an example, its operation is shown in Table 1. Analysis method: Two-point endpoint method, that is, the dosage of reagent R1 and reagent R2 is 150ul and 50ul respectively, and the sample volume is 3ul; 150ul reagent R1 is added to 3ul sample, and 50ulR2 is added after 3min at 37°C, that is, the point is read, and the reaction is read after 7min Another point; the main wavelength of the detection wavelength is 570nm.

[0049] Adopt this reagent and above-mentioned assay method, adopt the curve of the NGAL standard substance (self-made) of 6 kinds of different contents that Hitachi 7170 biochemical analyzer records (such as figure 1 Shown), each point represents a content of the reference standard, wherein the X-axis represents the NGAL content (ng / ml); Y-axis represents the absorbance.

[0050] Table 1 ...

Embodiment 3

[0052] Embodiment 3: Correlation test of detection reagent

[0053] Use the inventive reagent of this method (the specific formula is the same as embodiment 1) and the NGAL latex-enhanced immune turbidimetric reagent of a well-known company as a contrast reagent, adopt automatic 7170 full-automatic biochemical analyzer to test 50 parts of human urine (normal and abnormal specimens) by The respective parameters were measured at the same time, and the correlation analysis was carried out on the measured values. Measure according to the parameters in the above-mentioned NGAL determination method, and the determination results are shown in figure 2 , X, Y axis are measured value (the content ng / mL of NGAL), by figure 2 The results show that the correlation between the two reagents is R 2 =0.9914, the regression equation is y=0.9855x+0.9299. The results show that this reagent has a good correlation with a well-known reagent in the determination of patient urine, and has good...

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Abstract

The invention provides a latex enhanced immunoturbidimetry NGAL detection kit. The latex enhanced immunoturbidimetry NGAL detection kit comprises a reagent R1, a reagent R2 and a standard substance; the reagent R1 comprises a buffer solution, a surfactant, a chelating agent, an accelerator and an antiseptic; the reagent R2 comprises the buffer solution, NGAL antibody-coated latex particles, and an unrelated protein; and the standard substance is a solution containing 6 NGAL recombinant protein concentration gradients. Compared with kits in the prior art, the neutral granulocyte gelatinase related lipid transporter content detection kit used on a fully automatic biochemical analyzer has the advantages of rapid determination of the content of the NGAL in urine or blood plasma, guarantee of the sensitivity and the linearity, and improved sensitivity, accuracy and linearity of the determination result.

Description

technical field [0001] The invention belongs to the detection field of neutrophil gelatinase-related lipid carrier egg content in human body, relates to a latex-enhanced immune turbidimetric NGAL detection kit, and specifically relates to a method for using the detection kit in detecting NGAL content in a human body . 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 injury. 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 NGAL is a sensitive marker of early acute kidney injury. [0003] NGAL is a distinct protein combined with gelatinase discovered in animal neutrophils in 1993, with a rel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/54313G01N33/68
Inventor 李伟奇赵小凤房君江张秀文林清玉
Owner 上海睿康生物科技有限公司
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