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 overcome these limitations, we designed a new 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 1-10000 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.

中文翻译：

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CRISPR 介导的即时检测，用于检测溃疡性结肠炎动物模型中的粘膜钙卫蛋白

炎症性肠病 (IBD) 是一种与胃肠道炎症相关的慢性疾病，会导致一系列虚弱症状。粪便钙卫蛋白是溃疡性结肠炎 (UC) 的既定生物标志物，溃疡性结肠炎是主要的 IBD 疾病之一，可指示消化道炎症的存在和严重程度。酶联免疫吸附测定 (ELISA) 作为粪便钙卫蛋白检测的金标准方法，在现场护理环境中既耗时又不切实际。此外，从患者身上获取粪便样本具有挑战性并且阻碍了纵向监测。为了克服这些限制，我们设计了一种检测钙卫蛋白的新方法，该方法利用成簇规则间隔短回文重复序列 (CRISPR)/Cas 技术。我们成功开发了一种便携式管式 CRISPR/Cas 检测方法，用于钙卫蛋白的即时检测。该测定使用荧光和比色分析技术，显示检测范围为 1-10000 ng/mL（超过 4 个对数单位）。通过对 UC 麻醉临床前啮齿动物模型中获得的粘膜样本进行测量，进一步验证了所建立的测定方法，与健康对照动物相比，UC 大鼠样本中检测到的钙卫蛋白浓度高 2-3 倍。这种即时检测可以为通过粘膜样本检测诊断和监测 IBD 提供快速、精确且用户友好的方法。