Point-of-Care Multiplex Detection of Respiratory Viruses,ACS Sensors

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Point-of-Care Multiplex Detection of Respiratory Viruses
ACS Sensors ( IF 8.2 ) Pub Date : 2024-08-05 , DOI: 10.1021/acssensors.4c00992
Jongwon Lim _{1,

2} , Katherine Koprowski _{1,

2} , Robert Stavins ₃ , Nhat Xuan ₄ , Trung-Hieu Hoang ₄ , Janice Baek _{2,

5} , Victoria Kindratenko _{1,

2} , Liliana Khaertdinova ₁ , Alicia Yeun Kim ₁ , Minh Do ₄ , William P King _{3,

4,

5} , Enrique Valera _{1,

2,

6} , Rashid Bashir _{1,

2,

3,

4,

5,

6,

7,

8}

Affiliation

The COVID-19 pandemic, in addition to the co-occurrence of influenza virus and respiratory syncytial virus (RSV), has emphasized the requirement for efficient and reliable multiplex diagnostic methods for respiratory infections. While existing multiplex detection techniques are based on reverse transcription quantitative polymerase chain reaction (RT-qPCR) and extraction and purification kits, the need for complex instrumentation and elevated cost limit their scalability and availability. In this study, we have developed a point-of-care (POC) device based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) that can simultaneously detect four respiratory viruses (SARS-CoV-2, Influenza A, Influenza B, and RSV) and perform two controls in less than 30 min, while avoiding the use of the RNA extraction kit. The system includes a disposable microfluidic cartridge with mechanical components that automate sample processing, with a low-cost and portable optical reader and a smartphone app to record and analyze fluorescent images. The application as a real point-of-care platform was validated using swabs spiked with virus particles in nasal fluid. Our portable diagnostic system accurately detects viral RNA specific to respiratory pathogens, enabling deconvolution of coinfection information. The detection limits for each virus were determined using virus particles spiked in chemical lysis buffer. Our POC device has the potential to be adapted for the detection of new pathogens and a wide range of viruses by modifying the primer sequences. This work highlights an alternative approach for multiple respiratory virus diagnostics that is well-suited for healthcare systems in resource-limited settings or at home.

中文翻译：

呼吸道病毒的即时多重检测

除了流感病毒和呼吸道合胞病毒 (RSV) 的共存之外，COVID-19 大流行也强调了对呼吸道感染高效、可靠的多重诊断方法的需求。虽然现有的多重检测技术基于逆转录定量聚合酶链反应 (RT-qPCR) 以及提取和纯化试剂盒，但对复杂仪器的需求和高昂的成本限制了其可扩展性和可用性。在这项研究中，我们开发了一种基于逆转录环介导等温扩增（RT-LAMP）的即时检测（POC）设备，可以同时检测四种呼吸道病毒（SARS-CoV-2、甲型流感、乙型流感）和 RSV），并在 30 分钟内执行两个对照，同时避免使用 RNA 提取试剂盒。该系统包括一个带有机械部件的一次性微流体盒，可自动处理样品，并配有低成本便携式光学读取器和用于记录和分析荧光图像的智能手机应用程序。该应用程序作为一个真正的即时护理平台，使用在鼻液中掺有病毒颗粒的拭子进行了验证。我们的便携式诊断系统可准确检测呼吸道病原体特异的病毒 RNA，从而实现双重感染信息的反卷积。每种病毒的检测限是使用化学裂解缓冲液中掺入的病毒颗粒来确定的。我们的 POC 设备有潜力通过修改引物序列来适应新病原体和多种病毒的检测。这项工作强调了多种呼吸道病毒诊断的替代方法，该方法非常适合资源有限的环境或家庭中的医疗保健系统。

更新日期：2024-08-05

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