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Magnetic bead time resolution fluorescence immunoassay quantitative determination NGAL (neutrophil celatinase associated lipocalin) kit

A time-resolved fluorescence and immunoquantitative technology, applied in biological testing, material inspection products, etc., can solve the problems of being unsuitable for use in grassroots community hospitals, low detection sensitivity, expensive equipment, etc., and achieve long fluorescence decay time and high quantum yield. The effect of low rate and small sample size

Inactive Publication Date: 2018-07-27
GUANGZHOU BIOKEY HEALTH TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although fluorescence immunochromatography and latex-enhanced immunoturbidimetry are simple in operation and short in detection time, they have disadvantages such as low detection sensitivity, poor accuracy, and poor repeatability; although chemiluminescence method has high sensitivity and accuracy, linear range Wide and other advantages, but there are disadvantages such as expensive equipment and long detection time, so it is not suitable for use in grassroots community hospitals

Method used

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  • Magnetic bead time resolution fluorescence immunoassay quantitative determination NGAL (neutrophil celatinase associated lipocalin) kit
  • Magnetic bead time resolution fluorescence immunoassay quantitative determination NGAL (neutrophil celatinase associated lipocalin) kit
  • Magnetic bead time resolution fluorescence immunoassay quantitative determination NGAL (neutrophil celatinase associated lipocalin) kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] In this example, the magnetic bead time-resolved fluorescence immunoquantitative detection kit for NGAL consists of reagent strips and NGAL calibrator.

[0028] Wherein, the shape of the reagent strip is fan-shaped, and 1 to 13 holes are arranged sequentially from left to right. Wells 1 and 2 are test wells, wells 3 and 4 are fluorescent marker wells, wells 6 and 7 are diluent wells, holes 8 and 9 are sample diluent wells, 12th are enhancement solution wells, and 5th and 10th wells are diluent wells. , 11,13 are preparatory holes. A magnet can be stored between the 1st and 2nd wells for magnetic separation experiments. The 1st and 2nd wells can store a liquid volume of 300 μl; the 3rd and 4th wells can be disassembled from the entire reagent strip to become independent components, which is convenient for fluorescent markers. Pack and store. The 3rd, 4th, and 5th wells can store a liquid volume of 400 μl; the 6th and 7th wells can store a liquid volume of 3000 μl; the ...

Embodiment 2

[0060] The making and detection of embodiment 2 reagent strips

[0061] The semi-finished reagent strips in this example are assembled through the following procedures: 50 μL NGAL immunomagnetic beads, 100 μL europium-labeled NGAL antibody, 2000 μL cleaning solution, 2000 μL cleaning solution, 400 μL enhancement solution, and then seal it with a sealing film machine, and the sealing film is coated with product information identifications that can be scanned and identified by an automatic fluorescence detection analyzer, including enterprise standard curves, batches, production dates, and expiration dates. The finished reagent strips, packaged NGAL calibrator and other accessories are assembled into a kit.

[0062] Sample testing:

[0063] ①Add sample:

[0064] Put the sample or calibrator to be tested into the loading system of the automatic instrument, insert the reagent strip into the reagent strip slot, and the instrument will automatically identify the product informatio...

Embodiment 3

[0075] The comparison of embodiment 3 and commercially available kit

[0076] Commercially available neutrophil gelatinase-associated apolipoprotein assay kit (chemiluminescence microparticle immunoassay) (Abbott Trading Co., Ltd.) and the method in Example 1 are used to detect samples (the concentration range of NGAL in the sample is 10 ~1500ng / mL) was repeated three times to verify the correctness of the detection results of the kit of the present invention, and the results are shown in Table 1 below.

[0077] Table 1 Sample test results

[0078]

[0079]

[0080] Compared with commercially available reagents, the deviation of this reagent is less than ±5%, and the detection result of this reagent is accurate and reliable.

[0081] Commercially available neutrophil gelatinase-associated apolipoprotein assay kit (chemiluminescent microparticle immunoassay) (Abbott Trading Co., Ltd.) and the reagents in Example 1 were used to detect 200 cases of clinical urine samples, ...

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Abstract

The invention discloses a magnetic bead time resolution fluorescence immunoassay quantitative determination NGAL kit. The NGAL kit comprises an immunomagnetic bead coating a NGAL monoclonal antibody,a NGAL standardized product solution, an europium-marked NGAL monoclonal antibody solution, washing liquid and enhancement liquid. The immunomagnetic bead coating the NGAL monoclonal antibody is a covalent conjugate of a superpara magnetic bead modified by a functional group and with the diameter being 1-3 microns and the NGAL monoclonal antibody. The kit has the high sensibility, the sensibilityof NGAL is 1ng / mL, and a blood serum (plasma) does not need to be diluted; the determination time is short, and a report can be resulted within 30 minutes; the demanding amount of the sample is less,and only 50 microliters are needed for one-time sample loading; and the kit is equipped with a full-automatic time resolution immune analysis meter, operation is easy, no artificial error exists, andlabor is saved.

Description

technical field [0001] The invention relates to the technical field of in vitro reagent detection, in particular to a magnetic bead time-resolved fluorescence immunoquantitative detection kit for NGAL. Background technique [0002] Acute kidney injury (Acute Kidney Injury, AKI) is a clinical syndrome caused by a sudden decline in renal function in a short period of time (hours to days) caused by various reasons. AKI is clinically frequent and has a high mortality rate. Epidemiological surveys show that the incidence rate is 5% to 7% among the total hospitalized patients, and 25% among the patients in the intensive care unit, of which the mortality rate is about 50%~ 80%. 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. [0003] Neutrophil Gelatinase-associated Lipocalin (Neutrophil Gelatinase-associated Lipoca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 李根平
Owner GUANGZHOU BIOKEY HEALTH TECH CO LTD
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