Chemiluminiscence quantitative detection for excrement calprotectin, and detection method and application thereof in intestinal health detection

Abstract

The invention discloses chemiluminiscence quantitative detection for excrement calprotectin, a detection method thereof and an application thereof in intestinal health detection. The chemiluminiscencequantitative detection adopts a detection reagent, and comprises an immune complex and an antibody detection system, wherein the immune complex comprises a calprotectin calibrator, a calprotectin control product, and a labeled calprotectin complex antibody formed by compounding a labeled first specific antibody, a labeled second specific antibody and a to-be-detected sample. The combination modeof the first specific antibody, the second specific antibody and calprotectin is as follows: one antibody is specifically combined with an amino acid epitope in an amino acid sequence of a calprotectin S100A8 peptide chain, and the other antibody is specifically combined with an amino acid epitope in an amino acid sequence of a calprotectin S100A9 peptide chain. The chemiluminiscence quantitativedetection for excrement calprotectin, the detection method thereof and the application thereof can precisely and quantitatively detect the content of calprotectin in a human excrement sample, have theadvantages of high sensitivity and good stability, and have high clinical auxiliary value for intestinal health detection.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a chemiluminescent quantitative detection of fecal calprotectin, a detection method thereof, and an application thereof in intestinal health detection. Background technique [0002] Inflammatory bowel disease (IBD) is a chronic, idiopathic intestinal inflammatory disease involving the ileum, rectum, and colon. Clinical manifestations include diarrhea, abdominal pain, and even bloody stools. This disease includes ulcerative colitis (ulcerative colitis, UC) and Crohn's disease (Crohn's disease, CD). Ulcerative colitis is a continuous inflammation of the colonic mucosa and submucosa. The disease usually involves the rectum first and gradually spreads to the entire colon. Crohn's disease can involve the entire gastrointestinal tract and is a non-continuous full-thickness inflammation, most commonly involving the terminal ileum, colon, and perianal area. The etiology and pathoge...

AI Technical Summary

Problems solved by technology

However, colonoscopy and tissue biopsy have the following problems: it is an invasive examination, and the patient has a painful experience; the examination time is relatively long, and the whole process requires the active cooperation of the patient; in clinical work, some patients are difficult to accept and have poor compliance; for children In other words, they cannot tolerate the pain caused by colonoscopy, and it is difficult to cooperate with the examination; it is worth noting that for severe or fulminant patients, colonoscopy may induce toxic megacolon. Multiple colonoscopy examinations are contraindicated; in addition, colonoscopy cannot evaluate the inflammation of the intestinal lumen above the stenosis when the stenosis is complicated and the scope cannot pass through.

Among them, the representative product on the market using colloidal gold chromatography is the calprotectin detection kit of Xiamen Weizheng Biotechnology Co., Ltd., because the colloidal gold and the labeled protein are combined by electrostatic interaction, the combination is unstable, and the transaction is affected by acid and alkali. Factors and other external interference, so its detection sensitivity is low, and it is often used in qualitative or semi-quantitative detection, and cannot be accurately quantitatively detected; the representative product on the market using immunofluorescence chromatography is the calprotectin assay kit of Guangzhou Fengrun Biotechnology Co., Ltd. , because fluorescent light requires an external light source, detection in the direction perpendicular to the light source, molecules such as proteins and amino acids in biological samples will also produce background fluorescence, which will easily lead to problems such as high background and interference factors in the measurement

In addition, when fluorescence immunoassay is used for quantitative determination, it is necessary to select appropriate fluorescent reagents and sample processing methods to reduce the impact of non-specifically adsorbed proteins. The technology is complicated and the cost is high; the marketed product using enzyme-linked immunoassay is Buhlmann Laboratories of Germany AG’s calprotectin detection kit can quantitatively determine the content of calprotectin in feces, but the enzyme-linked immunoassay method requires cumbersome steps such as washing the plate, incubating, and adding chromogenic substrates, and the test process is affected by ambient temperature, etc. The impact is large, professional operation is required, and the test time is long

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