The accurate and sensitive detection of glucose from secretory clinical samples, such as tears and saliva, remains a great challenge. In this research, a novel ultrasensitive glucose detection method consisting of a glucose oxidase (GOx), pistol-like DNAzyme (PLDz), and CRISPR–Cas12a system is proposed. First, the oxidation of glucose catalyzed by GOx leads to the production of H₂O₂; the self-cleavage activity of PLDz is activated after recognition of the produced H₂O₂. The two procedures triggered by COx and PLDz play an important role in accurately identifying glucose and converting glucose signals to nucleic acids. The obtained PLDz fragments can be recognized by the Cas12 enzyme and thus activate the trans-cleavage activity of the Cas12a enzyme. Finally, the surrounding reporter probes are cut by the Cas12a enzyme to produce fluorescence signals. In summary, an ultra-sensitive and specific fluorescence method has been developed for glucose detection from secretory clinical samples, which could potentially contribute to the noninvasive diagnosis of diabetes mellitus.

中文翻译：

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通过整合葡萄糖氧化酶偶联脱氧核糖核酸酶和 CRISPR-Cas12a 从眼泪和唾液中进行超灵敏葡萄糖检测

从泪液和唾液等分泌性临床样本中准确、灵敏地检测葡萄糖仍然是一个巨大的挑战。在本研究中，提出了一种由葡萄糖氧化酶 (GOx)、手枪样脱氧核糖核酸酶 (PLDz) 和 CRISPR-Cas12a 系统组成的新型超灵敏葡萄糖检测方法。首先，GOx催化葡萄糖氧化产生H ₂ O ₂；PLDz的自切割活性在识别产生的H ₂ O ₂后被激活。COx 和 PLDz 触发的两个程序在准确识别葡萄糖和将葡萄糖信号转化为核酸方面发挥着重要作用。得到的 PLDz 片段可以被 Cas12 酶识别从而激活反式-Cas12a 酶的切割活性。最后，周围的报告探针被 Cas12a 酶切割以产生荧光信号。总之，已经开发出一种超灵敏和特异的荧光方法来检测分泌性临床样本中的葡萄糖，这可能有助于糖尿病的无创诊断。