Immuno-chromatography kit for quickly and quantitatively detecting calprotectin in excrement

A calprotectin, quantitative detection technology, applied in biological testing, measuring devices, analytical materials, etc., can solve the problems of unsuitable clinical rapid diagnosis, high requirements for testing equipment, poor repeatability, etc., to achieve reliable diagnosis results, improve The effect of detection stability and improved detection efficiency

Inactive Publication Date: 2016-08-10
江苏奥的特生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

ELISA technology has the following disadvantages: high requirements for detection equipment, high cost; many interference factors, poo

Method used

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  • Immuno-chromatography kit for quickly and quantitatively detecting calprotectin in excrement
  • Immuno-chromatography kit for quickly and quantitatively detecting calprotectin in excrement
  • Immuno-chromatography kit for quickly and quantitatively detecting calprotectin in excrement

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Composition of the Immunochromatographic Kit for Quantitative Detection of Fecal Calprotectin

[0032] combine Figure 1-3 Describe the structure of the immunochromatographic kit for the quantitative detection of fecal calprotectin. The immunochromatography kit includes a test strip, and the test strip includes a base plate 5, a sample pad 1 arranged on the base plate 5, a marker binding pad 2, a nitrocellulose membrane 3 and an absorbent pad 4, and the marker binding pad 2 packs Calprotectin monoclonal antibody A is labeled with colloidal gold particles, and the nitrocellulose membrane 3 contains parallel detection line T and quality control line C. The detection line T is formed by calprotectin monoclonal antibody B, and the quality control line C is formed by Goat anti-mouse IgG was formed, and calprotectin monoclonal antibodies A and B recognized different epitopes of calprotectin.

[0033] Calprotectin monoclonal antibody A was purchased from Zhuhai Bomei...

Embodiment 2

[0038] Example 2 Preparation of an immunochromatographic kit for quantitative detection of fecal calprotectin

[0039] In Example 1, the kit was prepared by the following method:

[0040] 1. Sample pad

[0041] The glass cellulose membrane was soaked in a surfactant buffer solution and dried at 37°C and a relative humidity of less than 20% to obtain a sample pad. The surfactant buffer solution is to add casein with a final concentration of 5% (mass percentage concentration), 5% (mass percentage concentration) of Polyvinylpyrrolidone 10, 1% (mass percentage concentration) in pH9.0 boric acid buffer solution It is formed after sodium cholate, 1.8% (mass percentage concentration) of RHODASURF ON-870 (SIGMA company reagent) and 0.02% (mass percentage concentration) sodium azide.

[0042] pH9.0 boric acid buffer solution: add 980 ml of ultrapure water to 6.18 g of boric acid to dissolve, then adjust the pH to 9.0 with 10 mol / L NaOH aqueous solution, and add ultrapure water to 100...

Embodiment 3

[0053] Example 3 The use method and effect of the immunochromatographic kit for quantitative detection of fecal calprotectin

[0054] 200 clinical stool samples were sampled with a stool specimen box, diluted and mixed with normal saline. In the sample hole 7 of the kit in Example 1, 3-4 drops of the treated sample were added dropwise, and after 10 minutes, the detection line T and the quality control line C were analyzed by an intelligent immune quantitative analyzer or an automatic stool analyzer. Optical density, calculate the concentration of fecal calprotectin in the processed sample according to the standard curve preset in the quantitative analyzer, the unit is ng / ml. Calculate the mass of fecal calprotectin per gram of fecal sample in ug / g.

[0055] The above-mentioned 200 clinical samples were detected by using the fecal calprotectin detection kit (ELISA) produced by Buhlmann Laboratories AG of Switzerland at the same time, and the correlation of the detection result...

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Abstract

The invention provides an immuno-chromatography kit for quickly and quantitatively detecting calprotectin in excrement and belongs to the technical field of clinical in-vitro diagnosis reagents. The kit includes: a test strip, which includes a substrate, and a sample pad, a marker combination pad, a nitrocellulose membrane and a water absorption pad which are all arranged on the substrate. The marker combination pad is coated with a calprotectin monoclonal antibody A marked by colloidal gold particles, colloidal silver particles, colloidal iron particles, magnetic particles, fuel particles, latex particles or fluorescent particles. The nitrocellulose membrane contains a detection line T and a quality control line C, wherein the detection line T is formed by a calprotectin monoclonal antibody B, and the quality control line C is formed by goat-anti-mouse IgG. The calprotectin monoclonal antibody A and the calprotectin monoclonal antibody B respectively recognize different epitopes of the calprotectin. The kit is convenient and quick, is simple in operation, has accurate result and is suitable for clinical quick detection.

Description

technical field [0001] The invention belongs to the technical field of clinical in vitro diagnostic reagents, in particular to an immunochromatographic kit for rapid quantitative detection of fecal calprotectin. Background technique [0002] Calprotectin is a heterozygous calcium-binding protein with a relative molecular mass of 36 000 Da. It is a heterodimer formed by the non-covalent combination of two heavy chains MRP14 and one light chain MRP8. Each chain can bind two calcium ions, has anti-protease activity in the presence of calcium ions, and has heat resistance due to the ability to chelate zinc ions, these properties make calprotectin in the intestinal lumen and the external environment It can remain stable for a long time without being destroyed by various enzymes and bacteria. [0003] Calprotectin is mainly derived from neutrophils and monocytes, and has a variety of biological functions. In vitro studies have shown that it has antibacterial properties, and the a...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/558G01N33/532
CPCG01N33/6893G01N33/532G01N33/558G01N33/577
Inventor 李降龙尹嘉
Owner 江苏奥的特生物技术有限公司
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