Kit for detecting calprotectin in human fecal sample and preparation method thereof

Abstract

The invention discloses a calprotein detection kit as well as a preparation method and a detection method thereof. The kit comprises a box body and a piece of detection test paper arranged in the box body, wherein the detection test paper comprises a base plate as well as a sample pad, a first conjugate pad, a second conjugate pad, a coating membrane and a piece of water absorbing paper which are sequentially connected from left to right and are arranged on the base plate; the first conjugate pad contains a Eu<3+> latex microsphere-labeled anti-mouse calprotein antibody, and the second conjugate pad contains a biotinylated anti-mouse calprotein antibody; and a detection line T and a quality control line C which are in parallel arrangement are arranged on the coating membrane from left to right, the detection line T is coated with immobilized streptavidin, and the quality control line C is coated with an immobilized goat anti-mouse IgG antibody. The calprotein detection kit is convenient and efficient in detection, high in detection sensitivity, strong in specificity, high in repeatability, objective and reliable and can quantitatively detect calprotein.

Description

technical field [0001] The invention relates to the technical field of detection kits for intestinal inflammatory disease marker proteins, in particular to a kit for detecting calprotectin in human feces samples and a preparation method thereof. Background technique [0002] Calprotectin is also called MRP8 / 14, L1, (p8,14), p34. It is composed of two heteromers of calgranulin A (MRP8) and calgranulin B (MRP14). Calprotectin is a calcium-binding protein secreted primarily by neutrophils and monocytes. Calprotectin in feces is a marker of neoplastic and inflammatory bowel disease. Elevated levels of calprotectin indicate recurrence of the corresponding disease, and very low levels of calprotectin in feces largely indicate the absence of organic bowel disease. In clinical practice, fecal calprotectin is often used to assess the degree of intestinal inflammation, monitor the status of patients with Crohn's disease, ulcerative colitis or polypectomy, and distinguish between in...

AI Technical Summary

Problems solved by technology

Its disadvantages are: 1. Enzyme-catalyzed reaction is greatly affected by temperature, so its detection process is limited by temperature control and cannot be applied to field use

3. The whole immune reaction process takes a long time, generally more than 1.5h; washing is required during the reaction process, and the operation is cumbersome

[0004] In the immunochromatography method, the marker substances of colloidal gold and colored latex are mainly judged by visual observation without any equipment assistance, but the sensitivity of this detection method is not high, the repeatability is poor, the detection range is narrow, and quantitative detection is not possible. It is only used for qualitative items that do not require high detection sensitivity

Fluorescent microspheres are latex microspheres mixed with organic fluorescent molecules. Due to the use of fluorescent labels, the detection sensitivity is 10-100 times higher than that of colloidal gold. Strong fluorescence background interference, especially the reflection effect of the nitrocellulose membrane (NC membrane) used in chromatography on the excited fluorescence when it is in a wet state, these interferences limit the detection sensitivity and specificity of fluorescent microspheres

Since the Stokes shift between the excitation spectrum and the emission spectrum of most organic fluorescent molecules is small, a filter is required to filter the signal of the excitation light in the detection, which greatly increases the manufacturing cost of the detection instrument, making the detection equipment more expensive and limited. wide application of the technology

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