A CRISPR mediated point‐of‐care assay for the detection of mucosal calprotectin in an animal model of ulcerative colitis

IF 6.1 2区 医学 Q1 ENGINEERING, BIOMEDICAL
Selena Chia, Tianruo Guo, Ewa M. Goldys, Sophie C. Payne, Wenlu Duan, Nigel H. Lovell, Mohit N. Shivdasani, Fei Deng
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Abstract

Inflammatory bowel disease (IBD) is a chronic disorder associated with inflammation in the gastrointestinal tract, leading to a range of debilitating symptoms. Fecal calprotectin is an established biomarker for ulcerative colitis (UC), one of the main IBD diseases, which provides indications of the presence and severity of inflammation in the digestive tract. Enzyme‐Linked Immunosorbent Assay (ELISA) as a gold standard approach for fecal calprotectin detection is time‐consuming and impractical in point‐of‐care settings. Moreover, obtaining fecal samples from patients is challenging and inhibits longitudinal monitoring. To address these specific problems, we have developed a novel approach for detecting calprotectin which leverages clustered regularly interspaced short palindromic repeats (CRISPR)/Cas technology. We successfully developed a portable tube‐based CRISPR/Cas assay for point‐of‐care testing of calprotectin. This assay showed a detection range from 1 to 10,000 ng/ml (over 4 log units), using both fluorescent and colorimetric analytical techniques. The established assay was further validated through measurements in mucosal samples obtained in an anesthetised preclinical rodent model of UC, with 2–3 times higher calprotectin concentration detected in UC rat samples compared to that of healthy control animals. This point‐of‐care test may provide a rapid, precise, and user‐friendly approach for the diagnosis and monitoring of IBD through mucosal sample testing.
在溃疡性结肠炎动物模型中检测粘膜钙粘蛋白的 CRISPR 介导的护理点检测法
炎症性肠病(IBD)是一种与胃肠道炎症有关的慢性疾病,会导致一系列使人衰弱的症状。粪便钙粘蛋白是溃疡性结肠炎(UC)(主要的 IBD 疾病之一)的一种公认生物标记物,可显示消化道炎症的存在和严重程度。酶联免疫吸附试验(ELISA)是检测粪便钙粘蛋白的金标准方法,但这种方法耗时较长,在护理点环境中并不实用。此外,从患者体内获取粪便样本也具有挑战性,不利于进行纵向监测。为了解决这些具体问题,我们开发了一种新型方法来检测钙黏蛋白,该方法利用了聚类规则间隔短回文重复序列(CRISPR)/Cas 技术。我们成功开发了一种基于 CRISPR/Cas 的便携式试管检测法,用于对犊牛蛋白进行床旁检测。利用荧光和比色分析技术,该检测方法的检测范围为 1 至 10,000 纳克/毫升(超过 4 个对数单位)。通过对麻醉的临床前啮齿动物 UC 模型粘膜样本进行测量,进一步验证了所建立的检测方法,在 UC 大鼠样本中检测到的钙粘蛋白浓度是健康对照动物的 2-3 倍。这种床旁检测法可为通过粘膜样本检测诊断和监测肠道疾病提供一种快速、精确和用户友好的方法。
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来源期刊
Bioengineering & Translational Medicine
Bioengineering & Translational Medicine Pharmacology, Toxicology and Pharmaceutics-Pharmaceutical Science
CiteScore
8.40
自引率
4.10%
发文量
150
审稿时长
12 weeks
期刊介绍: Bioengineering & Translational Medicine, an official, peer-reviewed online open-access journal of the American Institute of Chemical Engineers (AIChE) and the Society for Biological Engineering (SBE), focuses on how chemical and biological engineering approaches drive innovative technologies and solutions that impact clinical practice and commercial healthcare products.
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