The gold standard for diagnosing infectious diseases is culture-based identification of bacterial pathogens, which is time-consuming and labour-intensive. Current advances in molecular diagnostics and microfluidic technologies have made the rapid detection of bacteria or viruses in clinical specimens possible. However, the need for rapid, sensitive and multiplex detection of pathogens in a "sample-in and answer-out" manner has not been fully satisfied. In this study, a self-contained and fully integrated fluidic cassette and its supporting analyser were constructed for multiplex detection of bacteria to accelerate the diagnosis of urinary tract infections (UTIs). The fully integrated cassette contains all the necessary components and reagents for bacterial analysis. All of the bacterial analysis processes, including bacterial lysis, magnetic silica bead-based DNA extraction, DNA elution and multiplex loop-mediated amplification (LAMP), are automatically conducted in the cassette. This cassette was successfully applied for the detection of four major pathogenic bacteria in UTIs, i.e., Escherichia coli, Proteus mirabilis, Salmonella typhimurium and Staphylococcus aureus. The first three were successfully detected with a limit of detection (LoD) of 1 colony-forming unit (CFU) μL-1 and the last was with a LoD of 10 CFU μL-1 in urine samples, demonstrating that the cassette has similar sensitivity compared to that of the manual protocol, which is lower than that required by UTIs. The turnaround time for this cassette-based sample-to-answer system was approximately 100 minutes, and the detection is sensitive, fully automated, and accurate, demonstrating the potential to be a useful diagnostic tool for UTIs.

中文翻译：

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一个独立且完全集成的流体盒系统，用于细菌核酸的多重核酸检测。

诊断传染病的金标准是基于培养的细菌病原体鉴定，这既费时又费力。分子诊断和微流技术的最新进展使得快速检测临床标本中的细菌或病毒成为可能。但是，尚未完全满足对以“样本输入和答案输出”方式快速，灵敏和多重检测病原体的需求。在这项研究中，构建了一个独立且完全集成的流体盒及其支持的分析仪，用于细菌的多重检测，以加快尿路感染（UTI）的诊断。完全集成的试剂盒包含用于细菌分析的所有必需组件和试剂。所有细菌分析过程，包括细菌裂解，在盒中自动进行基于磁性硅胶珠的DNA提取，DNA洗脱和多重环介导的扩增（LAMP）。该盒已成功应用于尿路感染中四种主要致病菌的检测，即大肠杆菌，变形杆菌，鼠伤寒沙门氏菌和金黄色葡萄球菌。在尿液样品中成功检测到前三个，检出限（LoD）为1个菌落形成单位（CFU）μL-1，最后一个为10 CFUμL-1，表明该盒具有相似的灵敏度与手动协议相比，它比UTI所要求的要低。此盒式样品采集系统的周转时间约为100分钟，并且检测灵敏，全自动，准确，