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Device for combined quantitative detection of Ungal and UCR based on up-conversion luminescence technology and its preparation method

A quantitative detection and technology technology, applied in the field of immunological detection, to achieve the effect of convenient detection process, fast detection process and less sample consumption

Active Publication Date: 2017-10-03
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has great application potential in laboratory research and clinical testing for early diagnosis of renal injury and other diseases such as acute bacterial infection

Method used

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  • Device for combined quantitative detection of Ungal and UCR based on up-conversion luminescence technology and its preparation method
  • Device for combined quantitative detection of Ungal and UCR based on up-conversion luminescence technology and its preparation method
  • Device for combined quantitative detection of Ungal and UCR based on up-conversion luminescence technology and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Combined quantitative detection of total uNGAL and uCr device composition based on up-conversion luminescence technology

[0047] 1) 1 part of sample diluent lyophilized powder (reagent Ⅰ)

[0048] 2) 1 tube of standard D solution freeze-dried powder (Reagent Ⅱ)

[0049] 3) 1 tube of UCP-labeled anti-NGAL antibody against three molecular forms and UCP-labeled Cr solution (reagent Ⅲ)

[0050] 4) Class A immunochromatographic test strips 96 pieces

[0051] 5) Class A immunochromatographic test strip supporting test strip holder 1

[0052] 6) One piece of 96-well plate, one piece of sealing film for 96-well plate

Embodiment 2

[0053] Example 2: Combined quantitative detection of total NGAL and urinary creatinine device composition based on up-conversion luminescence technology

[0054] 1) 1 part of sample diluent lyophilized powder (reagent Ⅰ)

[0055] 2) 1 tube of standard D solution freeze-dried powder (Reagent Ⅱ)

[0056] 3) Type B immunochromatographic test strips 100 pieces

Embodiment 3

[0057] Example 3: Preparation and use of a device based on up-conversion luminescence technology for combined quantitative detection of total uNGAL and uCr

[0058] 1) Preparation of device

[0059] a) Preparation of sample diluent lyophilized powder (reagent Ⅰ)

[0060] Sample diluent 20mL, composed of 100mmol / L HEPES, pH 7.2, 200mmol / L NaCl, 0.25% (vol / vol) Tween-20, 0.5% (wt / vol) BSA; after sufficient magnetic stirring, use 0.45μm The filter was sterilized by filtration and insoluble impurities were removed, and then freeze-dried; stored at room temperature.

[0061] b) Preparation of standard D solution freeze-dried powder (reagent Ⅱ)

[0062] The pH= After the 10mmol / L PBS solution of 7.4 is fully stirred by magnetic force, use a 0.22μm filter to filter and sterilize and remove insoluble impurities, then freeze-dry; store at room temperature.

[0063] c) Preparation of UCP-labeled anti-NGAL antibody against three molecular forms and 1 tube of UCP-labeled Cr solution (...

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Abstract

The invention discloses an immunochromatographic device for jointly quantitatively detecting urinary neutrophil gelatinase associated lipocalin (uNGAL) in a disease marker and urinary creatinine (uCr) serving as a dilution influence correcting factor for urine based on up-converting phosphor technology (UPT) and a preparation method of the device, and belongs to the technical field of immunodetection. The immunochromatographic device is prepared from sample diluent freeze-dried powder, NGAL (Neutrophil Gelatinase Associated Lipocalin) and Cr (Creatinine) standard substance freeze-dried powder and an immunochromatographic test paper strip, wherein the immunochromatographic test paper strip consists of a lining board, a sample pad or a UCP (Up-Converting Phosphor) conjugate pad, an analytical film and an absorbent pad; two parallel detecting lines T1 and T2 and a quality control line C are arranged on the analytical film; the two detecting lines T1 and T2 encrust an anti-NGAL antibody and an anti-Cr antibody respectively; the quality control line encrusts an antibody, which comes from animal immune globulin, of an anti-UCP labeled antibody; the test paper strip can quantify the uNGAL and the uCr by using a UCP readout instrument. The immunochromatographic device can be used for jointly quantifying the uNGAL and the uCr in different molecular forms, and has the advantages of quickness, simpleness, convenience and the like. After the uNGAL is corrected by the uCr, the capacity, for the early diagnosis of diseases of renal injury and the like, of the NGAL can be improved, and therefore, the immunochromatographic device has enormous scientific-research and clinical-application value.

Description

technical field [0001] The invention belongs to the technical field of immunological detection, and in particular relates to the combined quantitative detection of disease biomarkers urinary neutrophil gelatinase-associated apolipoprotein (uNGAL) and urine dilution effect correction factor urinary creatinine based on up-conversion luminescence technology (UPT) (uCr) immunochromatography device and preparation method thereof. Background technique [0002] Neutrophil gelatinase associated lipocalin (NGAL), also known as human neutrophil lipocalin (HNL), was discovered in the second granule of neutrophils in the early 1990s A glycoprotein of lipocalin, a member of the lipocalin superfamily, has a similar spatial structure to other lipocalins (J.Biol.Chem.1993, 268:10425-10432; Scand.J.Clin.Lab Invest. 1994, 54:365-376). NGAL exists in various molecular forms such as monomer, homodimer and heterodimer with MMP9, and different molecular forms of NGAL have differential correlati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/92G01N33/70G01N33/577G01N33/558G01N21/63
CPCG01N21/63G01N33/558G01N33/577G01N33/70G01N33/92G01N2333/775
Inventor 蔡林君陈雷杨津赵冰姜春来孔维
Owner JILIN UNIV