DNAzyme/珠状荧光法检测冷却塔水样中的嗜肺军团菌

IF 3.4 Q2 CHEMISTRY, ANALYTICAL
Shuwen Qian, Dr. Erin M. McConnell, Meghan Rothenbroker, Jimmy Gu, Simina Alungulesa, Louis Godbout, Prof. Yingfu Li
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引用次数: 0

摘要

嗜肺军团菌是致命的水媒疾病军团病的病原体,这种疾病通常通过冷却塔水样中受污染的飞沫传播。缺乏有效的检测方法对嗜肺乳杆菌的疫情控制提出了挑战。此前,一种名为LP1的rna切割DNAzyme被报道专门针对嗜肺乳杆菌。本研究通过链亲和素-生物素相互作用将LP1固定在琼脂糖珠上,建立了一种基于珠状荧光检测嗜肺乳杆菌的方法。该方法在不同的冷却塔水样中表现出良好的稳定性和功能性。为了改善嗜肺乳杆菌在实际样品中的监测,采用溶菌酶处理来增强嗜肺乳杆菌的识别。在没有细胞培养和信号放大步骤的情况下,该dnazyme蛋白头检测方法的检出限可达103 CFUs。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Detecting Legionella pneumophila in Cooling Tower Water Samples with a DNAzyme/Bead-Based Fluorescence Assay

Detecting Legionella pneumophila in Cooling Tower Water Samples with a DNAzyme/Bead-Based Fluorescence Assay

Legionella pneumophila is the causative agent behind the deadly waterborne disease Legionnaires’, which is commonly transmitted by the spread of contaminated droplets from cooling tower water samples. The lack of effective detection methods presents a challenge for L. pneumophila outbreak control. Previously, an RNA-cleaving DNAzyme called LP1 was reported to specifically target L. pneumophila. In this study, LP1 was immobilized onto agarose beads via streptavidin-biotin interaction to develop a bead-based fluorescence assay for L. pneumophila detection. This bead-based assay demonstrated excellent stability and functionality in various cooling tower water samples. To improve L. pneumophila monitoring in real-world samples, a lysozyme treatment was used to enhance L. pneumophila recognition. The limit of detection of this DNAzyme-based bead assay can reach 103 CFUs in cell-spiked cooling tower water samples without cell culturing or signal amplification steps.

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