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Kit for detecting calprotectin in human feces and preparation method thereof

A technology of calprotectin and kits, applied in the biological field, can solve problems such as cumbersome operation, susceptibility to environmental interference, and long extraction time by shaking

Inactive Publication Date: 2018-07-27
SHENZHEN BLOT BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the German Buhlmann Laboratories AG calprotectin kit using ELISA can accurately test the content of calprotectin in feces, but ELISA needs to wash the plate, incubate, add enzymes and substrates, and the temperature environment is strict. , the operation is cumbersome and cannot be tested on site, and the test time is long
The calprotectin detection kit prepared by Xiamen Weizheng Biotechnology Co., Ltd. uses the colloidal gold method to test the content of calprotectin. Since the colloidal gold particles and the labeled protein are combined by electrostatic interaction, the combination is unstable and susceptible to environmental interference. Low sensitivity, unable to quantitatively detect
Moreover, before these existing detection kits detect calprotectin in feces, the extraction liquid is usually required to dissolve the feces and shake for more than 10 minutes, and the shaking extraction time is longer

Method used

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  • Kit for detecting calprotectin in human feces and preparation method thereof
  • Kit for detecting calprotectin in human feces and preparation method thereof
  • Kit for detecting calprotectin in human feces and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] The preparation of embodiment 1 kit of the present invention

[0048] Use thermo company EZ- Sμlfo-NHS-LC-Biotin Biotinylation Kit Bovine serum albumin (BSA) is biotinylated, purified by dialysis to obtain the Bi-BSA conjugate and temporarily stored for future use.

[0049] 1. Preparation of the sample pad: soak the sample pad in 0.25-0.5M Tris-HCl buffer solution containing 5-10% trehalose for 5-10 minutes, dry at 37 degrees for 2 hours and store in a sealed manner.

[0050] 2. Preparation of the reaction membrane: Coat the biotin-bovine serum albumin conjugate on the nitrocellulose membrane as the T line, and coat the diluted goat anti-mouse IgG antibody on the nitrocellulose membrane as the C line , dried at 37°C for 16-22h, sealed and stored for later use.

[0051] 3. Preparation of Conjugate Pad 1

[0052] 3.1 Preparation of anti-calprotectin antibody biotinylated conjugates

[0053] use thermo company Sμlfo-NHS-LC-Biotin Biotinylation Kit biotinylated anoth...

Embodiment 2

[0071] Embodiment 2 Preparation of kit of the present invention

[0072] In the preparation of the detection test strip in the kit, the preparation of the binding pad 1 in step 3, the sealing agent used is 2wt% gelatin (w / w), 2wt% NH 2 - 0.5M MES buffer solution of PEG and 0.5wt% trehalose, other steps are the same as in Example 1.

Embodiment 3

[0073] Embodiment 3 Preparation of kit of the present invention

[0074] The formula of feces extraction buffer in this kit is: 4.0M urea, 0.025M CaCl 2 , 0.25M Tris-HCl buffer solution with a pH of 8.0 of 0.5wt% SDS, 10mM EDTA, 0.5M citric acid, 0.05wt% sodium azide and 5wt% BSA, and other steps were the same as in Example 1.

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Abstract

The invention relates to the biological technical field, and in particular, relates to a kit for detecting calprotectin in human feces and a preparation method thereof. The kit uses a specific microsphere conjugate sealing reagent instead of a traditional sealant, and can accurately, quantitatively, quickly and sensitively detect the calprotectin in the human feces.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit and a preparation method for detecting calprotectin in human feces. Background technique [0002] Calprotectin is composed of two heavy chains with a molecular weight of 14kD and a light chain with a molecular weight of 8kD. It is composed of a calcium-binding protein heterotrimer linked by a covalent bond. It is an important protein in the cytoplasm of neutrophils and activated macrophages. protein, levels are increased in many inflammatory conditions. As a marker of intestinal inflammation, it can clearly distinguish the organic inflammatory disease IBD from the functional disease IBS, and can be used as an active index to evaluate inflammatory bowel disease, evaluate the mucosa of inflammatory bowel disease, and evaluate the effect of drug therapy , cure and predict disease recurrence. Routine endoscopic examination and biopsy histological analysis are invasive examination...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/558G01N33/533G01N21/64
CPCG01N21/6486G01N33/533G01N33/558G01N33/6893G01N2800/065G01N2333/4727G01N33/582G01N33/54387
Inventor 肖川张大准张永顶马伟民
Owner SHENZHEN BLOT BIOTECH
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