Aspergillus fumigatus has widely evolved resistance to the most commonly used class of antifungal chemicals, the azoles. Current methods for identifying azole resistance are time-consuming and depend on specialized laboratories. There is an urgent need for rapid detection of these emerging pathogens at point-of-care to provide the appropriate treatment in the clinic and to improve management of environmental reservoirs to mitigate the spread of antifungal resistance. Our study demonstrates the rapid and portable detection of the two most relevant genetic markers linked to azole resistance, the mutations TR34 and TR46, found in the promoter region of the gene encoding the azole target cyp51A. We developed a lab-on-a-chip platform consisting of: (i) tandem-repeat loop-mediated isothermal amplification; (ii) state-of-the-art complementary metal-oxide-semiconductor microchip technology for nucleic acid amplification detection; and (iii) a smartphone application for data acquisition, visualization, and cloud connectivity. Specific and sensitive detection was validated with isolates from clinical and environmental samples from 6 countries across 5 continents, showing a lower limit of detection of 10 genomic copies per reaction in less than 30 min. When fully integrated with a sample preparation module, this diagnostic system will enable the detection of this ubiquitous fungus at the point-of-care, and could help to improve clinical decision making, infection control, and epidemiological surveillance.

中文翻译：

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使用芯片实验室诊断系统快速检测临床和环境分离物中的耐唑类烟曲霉。

烟曲霉对最常用的一类抗真菌化学物质——唑类具有广泛的耐药性。目前识别唑类耐药性的方法非常耗时，并且依赖于专门的实验室。迫切需要在护理点快速检测这些新出现的病原体，以便在临床上提供适当的治疗，并改善对环境水库的管理，以减轻抗真菌耐药性的传播。我们的研究表明，在编码唑靶cyp的基因的启动子区域中发现了与唑类抗性相关的两个最相关的遗传标记物 TR34 和 TR46 突变的快速和便携式检测51A。我们开发了一个芯片实验室平台，包括：（i）串联重复环介导的等温扩增；(ii) 用于核酸扩增检测的最先进的互补金属氧化物半导体微芯片技术；(iii) 用于数据采集、可视化和云连接的智能手机应用程序。对来自 5 大洲 6 个国家的临床和环境样本的分离株进行了特异性和灵敏检测验证，显示每个反应在 30 分钟内检测到 10 个基因组拷贝的下限。当与样品制备模块完全集成时，该诊断系统将能够在护理点检测这种无处不在的真菌，​​并有助于改善临床决策、感染控制和流行病学监测。