Here a triple functional sensing chip was created for L. monocytogenes detection by integrating three biomarkers (Listeriolysin O (LLO) at protein level, hly gene at genetic level, and acetoin at metabolic level). Liposome encapsulated catechol was used for LLO detection via LLO pore-forming ability. hly gene was specifically captured by using a thiolated capture probe on nanoporous gold (NPG). As an electroactive label, methylene blue was embedded in double-stranded structures to generate an electrochemical signal for hly detection. Combined with the electrocatalysis of NADH by NPG, the acetoin detection was achieved by measuring the consumption of NADH as a cofactor under acetoin reductase catalysis. Importantly, the L. monocytogenes detection results obtained by detecting three biomarkers using the chip can be mutually verified, which reduces the probability of false positives based on a single marker. Moreover, the detection time was reduced to about 90 min, making it a rapid and reliable tool for L. monocytogenes detection.

中文翻译：

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快速检测病原性李斯特菌的三功能感应芯片

在这里，通过整合三个生物标记（蛋白质水平的李斯特菌溶血素O（LLO），基因水平的hly基因和代谢水平的乙酰丁酸）创建了用于单核细胞增生李斯特氏菌检测的三功能感应芯片。脂质体包封的儿茶酚通过LLO成孔能力用于LLO检测。通过在纳米孔金（NPG）上使用硫醇化捕获探针特异性捕获hly基因。作为电活性标记，将亚甲基蓝嵌入双链结构中，以产生电化学信号进行快速检测。结合NPG对NADH的电催化作用，通过测量在乙酰辅酶还原酶催化下NADH作为辅因子的消耗量来实现乙酰辅酶的检测。重要的是，可以相互验证通过使用该芯片检测三种生物标志物而获得的单核细胞增生李斯特菌检测结果，这样可以减少基于单个标记的误报的可能性。此外，检测时间减少到约90分钟，使其成为单核细胞增生李斯特氏菌检测的快速可靠的工具。