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Label-free electrical sensing of bacteria in eye wash samples: A step towards point-of-care detection of pathogens in patients with infectious keratitis

机译:洗眼液样品中细菌的无标签电感应:迈向感染性角膜炎患者病原体现场检测的一步

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摘要

The diagnosis of keratitis is based on visual exam, tissue cytology, and standard microbial culturing to determine the type of the infectious pathogen. To prescribe appropriate therapy, it is important to distinguish between bacterial, fungal, and viral keratitis, as the treatments are quite different. Diagnosis of the causative organism has a substantial prognostic importance. Further, timely knowledge of the nature of the pathogen is also critical to adapt therapy in patients unresponsive to empiric treatment options, which occurs in 10% of all cases. Currently, the identification of the nature of the pathogen that causes keratitis is achieved via microbial culture screening, which is laboratory-based, expensive, and time-consuming. The most frequent pathogens that cause the corneal ulcers are P. aeruginosa and S. aureus. Here, we report a microchip for rapid (<1 h) detection of P. aeruginosa (6294), S. aureus (LAC), through on-chip electrical sensing of bacteria lysate. We evaluated the microchip with spiked samples of PBS with bacteria concentration between 101 to 108 CFU/mL. The least diluted bacteria concentration in bacteria-spiked samples with statistically significant impedance change was 10 CFU/ml. We further validated our assay by comparing our microchip results with the standard culture-based methods using eye washes obtained from 13 infected mice.
机译:角膜炎的诊断基于视觉检查,组织细胞学检查和标准微生物培养,以确定感染性病原体的类型。为了开出适当的治疗方法,重要的是区分细菌性,真菌性和病毒性角膜炎,因为治疗方法大不相同。致病生物的诊断具有重要的预后重要性。此外,及时了解病原体的性质对于适应对经验治疗选择无反应的患者的治疗也很关键,这种情况发生在所有病例中的10%。当前,通过基于实验室的,昂贵且费时的微生物培养筛选来鉴定引起角膜炎的病原体的性质。引起角膜溃疡的最常见病原体是铜绿假单胞菌和金黄色葡萄球菌。在这里,我们报告通过芯片上的细菌裂解物的电感应,快速(<1小时)检测铜绿假单胞菌(6294),金黄色葡萄球菌(LAC)的微芯片。我们用细菌浓度在10 1 至10 8 CFU / mL之间的PBS掺加样品评估了微芯片。具有显着阻抗变化的掺菌样品中的最低稀释细菌浓度为10 CFU / ml。我们通过使用从13只受感染的小鼠那里得到的洗眼液将微芯片结果与基于标准培养的方法进行比较,进一步验证了我们的测定方法。

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