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Mobile Platform for Multiplexed Detection and Differentiation of Disease-Specific Nucleic Acid Sequences, Using Microfluidic Loop-Mediated Isothermal Amplification and Smartphone Detection
Analytical Chemistry ( IF 7.4 ) Pub Date : 2017-09-05 00:00:00 , DOI: 10.1021/acs.analchem.7b02478
Weili Chen 1 , Hojeong Yu 1 , Fu Sun 1 , Akid Ornob 2 , Ryan Brisbin 3 , Anurup Ganguli 2 , Vinay Vemuri 1 , Piotr Strzebonski 1 , Guangzhe Cui 1 , Karen J. Allen 4 , Smit A. Desai 4 , Weiran Lin 1 , David M. Nash 5 , David L. Hirschberg 3, 6 , Ian Brooks 4 , Rashid Bashir 2 , Brian T. Cunningham 1, 2
Affiliation  

New tools are needed to enable rapid detection, identification, and reporting of infectious viral and microbial pathogens in a wide variety of point-of-care applications that impact human and animal health. We report the design, construction, and characterization of a platform for multiplexed analysis of disease-specific DNA sequences that utilizes a smartphone camera as the sensor in conjunction with a hand-held “cradle” that interfaces the phone with a silicon-based microfluidic chip embedded within a credit-card-sized cartridge. Utilizing specific nucleic acid sequences for four equine respiratory pathogens as representative examples, we demonstrated the ability of the system to utilize a single 15 μL droplet of test sample to perform selective positive/negative determination of target sequences, including integrated experimental controls, in approximately 30 min. Our approach utilizes loop-mediated isothermal amplification (LAMP) reagents predeposited into distinct lanes of the microfluidic chip, which when exposed to target nucleic acid sequences from the test sample, generates fluorescent products that when excited by appropriately selected light emitting diodes (LEDs), are visualized and automatically analyzed by a software application running on the smartphone microprocessor. The system achieves detection limits comparable to those obtained by laboratory-based methods and instruments. Assay information is combined with the information from the cartridge and the patient to populate a cloud-based database for epidemiological reporting of test results.

中文翻译:

使用微流体环介导的等温扩增和智能手机检测,用于疾病特定核酸序列的多重检测和鉴别的移动平台

需要新的工具来快速检测,鉴定和报告感染人类和动物健康的各种即时医疗应用中的传染性病毒和微生物病原体。我们报告了对疾病特异性DNA序列进行多重分析的平台的设计,构建和表征,该平台利用智能手机摄像头作为传感器,以及将手机与基于硅的微流控芯片对接的手持式“支架”嵌入信用卡大小的墨盒中。以四种马呼吸道病原体的特定核酸序列为代表,我们证明了系统利用单个15μL小滴测试样品对目标序列进行选择性阳性/阴性测定的能力,包括集成的实验对照,在大约30分钟内 我们的方法利用预沉积到微流体芯片,其暴露于目标从测试样品中的核酸序列时的不同车道环介导等温扩增(LAMP)的试剂,产生荧光产物,当通过适当地选择发光二极管(LED)激发,通过运行在智能手机微处理器上的软件应用程序可视化并自动分析。该系统所达到的检测极限可与基于实验室的方法和仪器所获得的极限相媲美。将检测信息与来自药筒和患者的信息相结合,以填充基于云的数据库,以便对检测结果进行流行病学报告。我们的方法利用预沉积到微流体芯片,其暴露于目标从测试样品中的核酸序列时的不同车道环介导等温扩增(LAMP)的试剂,产生荧光产物,当通过适当地选择发光二极管(LED)激发,通过运行在智能手机微处理器上的软件应用程序可视化并自动分析。该系统所达到的检测极限可与基于实验室的方法和仪器所获得的极限相媲美。将检测信息与来自药筒和患者的信息相结合,以填充基于云的数据库,以便对检测结果进行流行病学报告。我们的方法利用预沉积到微流体芯片,其暴露于目标从测试样品中的核酸序列时的不同车道环介导等温扩增(LAMP)的试剂,产生荧光产物,当通过适当地选择发光二极管(LED)激发,通过运行在智能手机微处理器上的软件应用程序可视化并自动分析。该系统所达到的检测极限可与基于实验室的方法和仪器所获得的极限相媲美。将检测信息与来自药筒和患者的信息相结合,以填充基于云的数据库,以便对检测结果进行流行病学报告。产生荧光产物,其在由适当地选择发光二极管(LED)激发,被可视化,并自动由在智能电话微处理器上运行的软件应用程序进行分析。该系统所达到的检测极限可与基于实验室的方法和仪器所获得的极限相媲美。将检测信息与来自药筒和患者的信息相结合,以填充基于云的数据库,以便对检测结果进行流行病学报告。产生荧光产物,其在由适当地选择发光二极管(LED)激发,被可视化,并自动由在智能电话微处理器上运行的软件应用程序进行分析。该系统所达到的检测极限可与基于实验室的方法和仪器所获得的极限相媲美。将检测信息与来自药筒和患者的信息相结合,以填充基于云的数据库,以便对检测结果进行流行病学报告。
更新日期:2017-09-06
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