Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD, USA.
Prompt Diagnostics LLC, Baltimore, MD, USA.
Sci Rep. 2023 Dec 9;13(1):21848. doi: 10.1038/s41598-023-47220-7.
The continued spread of Candida auris in healthcare facilities has increased the demand for widely available screening to aid in containment and inform treatment options. Current methods of detection can be unreliable and require bulky and expensive instruments to lyse and identify fungal pathogens. Here, we present a quick, low-cost, instrument-free method for lysis of C. auris suitable for streamlined sample processing with polymerase chain reaction (PCR) detection. Chemical, thermal, and bead beating lysis techniques were evaluated for lysis performance and compatibility with nucleic acid extraction and downstream PCR reactions. Using only 10 s of manual shaking with glass beads, this method demonstrated a limit of detection (LOD) of C. auris at 500 colony forming units per mL, a 20-fold improvement compared to the LOD without manual shaking, and a 60-fold reduction in time compared to common fungal lysis kits, all while maintaining repeatability and reproducibility across multiple users. This work highlights a simple method for increasing sensitivity and reducing turnaround time of PCR-based C. auris detection and exhibits promise for integration into point-of-care platforms towards real-time triage of colonized patients.
耳念珠菌在医疗机构中的持续传播增加了广泛筛查的需求,以帮助控制并提供治疗选择。目前的检测方法可能不可靠,并且需要庞大而昂贵的仪器来裂解和鉴定真菌病原体。在这里,我们提出了一种快速、低成本、无需仪器的方法,用于裂解耳念珠菌,适用于与聚合酶链反应(PCR)检测相结合的简化样本处理。我们评估了化学、热和珠击裂解技术的裂解性能以及与核酸提取和下游 PCR 反应的兼容性。仅使用玻璃珠手动摇晃 10 秒,该方法将耳念珠菌的检测限(LOD)降低到每毫升 500 个菌落形成单位,与没有手动摇晃相比提高了 20 倍,与常见的真菌裂解试剂盒相比,时间缩短了 60 倍,同时在多个用户之间保持了重复性和再现性。这项工作突出了一种简单的方法,可以提高基于 PCR 的耳念珠菌检测的灵敏度并减少周转时间,并有望整合到即时护理平台中,以实时对定植患者进行分诊。