An NGAL time-resolved fluoroimmunoassay nanometer immunochromatographic quantitative detection test paper strip and a preparing method thereof

A technology of time-resolved fluorescence and immunochromatography, applied in analytical materials, measuring devices, instruments, etc., to achieve good accuracy and precision, avoid overlapping, and slow down penetration.

Inactive Publication Date: 2016-04-27
XIAMEN YIKELISI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Up to now, there is no NGAL time-resolved fluorescent na

Method used

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  • An NGAL time-resolved fluoroimmunoassay nanometer immunochromatographic quantitative detection test paper strip and a preparing method thereof
  • An NGAL time-resolved fluoroimmunoassay nanometer immunochromatographic quantitative detection test paper strip and a preparing method thereof
  • An NGAL time-resolved fluoroimmunoassay nanometer immunochromatographic quantitative detection test paper strip and a preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0039] The preparation method of all kinds of solutions adopted in the above-mentioned method is as follows:

[0040] 0.05M MES buffer solution: Weigh 4.265g of MES (2-(N-morpholino)ethanesulfonic acid) salt, dissolve it in 80ml of water, adjust the pH value to 6.0 with 2M aqueous sodium hydroxide solution, and set the volume to 100ml. Obtain a 0.2M MES solution, which can be diluted four times with water before use.

[0041] 20mg / ml EDC solution: Weigh 0.020g EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide), dissolve it in 1ml of water, and obtain.

[0042] The boric acid buffer solution of pH7.5 of 0.05M: weigh 1.2366g borax (H 3 BO 3 ), add 80ml of water to dissolve completely, adjust the pH value to 7.5 with 2M aqueous sodium hydroxide solution, and set the volume to 100ml to obtain 0.2M boric acid buffer solution, which can be diluted four times with water for use.

[0043] Blocking solution: 40 mM glycine solution, adjusted to pH 7.5 with TB buffer, added BSA (bovi...

experiment example

[0048] Experimental example: Quantitative detection of samples

[0049] 1. Standard curve creation

[0050] 1.1 Take a set of NGAL calibration products, the specific values ​​are shown in the table below

[0051]

[0052] 1.2 Detection method

[0053] 1.2.1 Take 75 μl of calibrator and add it to the sample window in the chromatographic strip prepared in Example 1;

[0054] 1.2. After 210 minutes, use a time-resolved fluorescence quantitative analyzer to quantitatively detect the luminescence value.

[0055] 1.2.3 Each calibrator is tested three times, and the average value of T / C value is taken. The specific results are shown in the table below:

[0056]

[0057]

[0058] 1.3 Standard curve preparation

[0059] According to the above detection results, the logarithmic value of the T / C value is used as the X-axis, and the logarithmic value of the concentration is used as the Y-axis to perform linear regression to obtain a linear equation y=-0.054x 2 +1.0185x-4.92...

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Abstract

The invention discloses an NGAL time-resolved fluoroimmunoassay nanometer immunochromatographic quantitative detection test paper strip and a preparing method thereof. The test paper strip comprises a backing substrate plate. The backing substrate plate is provided with a nitrocellulose membrane, a sample combining pad and a water absorbing membrane. The sample combining pad and the water absorbing membrane are respectively laminated on two ends of the nitrocellulose membrane. The sample combining pad is covered with an NGAL monoclonal antibody labeled with a time-resolved fluorescent substance. The nitrocellulose membrane is provided with a detection line and a quality control line. The detection line is covered with an NGAL monoclonal antibody of another epitope. The quality control line is covered with goat anti-rat IgG. The test paper strip and the method are characterized by high sensitivity, high specificity, good stability, no radioactive contamination, and the like and can be widely applied to NGAL clinical immunoassay and scientific research.

Description

technical field [0001] The invention belongs to the technical field of immunochromatography detection, and in particular relates to a NGAL time-resolved fluorescence nano-immunochromatography quantitative detection test strip and a preparation method thereof. Background technique [0002] Acute kidney injury (Acute kidney injury, AKI) is a clinical syndrome caused by the decline of renal function in a short period of time due to various reasons. So far, AKI is still a very common and serious clinical problem. If AKI is treated in the early stages, the occurrence of renal failure can be greatly reduced. Therefore, early diagnosis of AKI has become a key factor in the treatment of renal disease. However, the lack of specific early clinical diagnostic indicators is the main obstacle to the treatment of the disease, resulting in high incidence and mortality of AKI. Human neutrophil gelatinase-associated apolipoprotein (NGAL, apolipoprotein-2) is a trace protein expressed by n...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/558
Inventor 卢志珍许元峰钟伟波
Owner XIAMEN YIKELISI MEDICAL TECH CO LTD
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