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基于 RT-LAMP 的分子诊断试剂盒用于快速丙型肝炎病毒检测。

RT-LAMP-Based Molecular Diagnostic Set-Up for Rapid Hepatitis C Virus Testing.

机构信息

Department of Electrical Engineering and Computer Science, Florida Atlantic University, Boca Raton, FL 33431, USA.

Asghar-Lab: Micro and Nanotechnology in Medicine, College of Engineering and Computer Science, Boca Raton, FL 33431, USA.

出版信息

Biosensors (Basel). 2022 May 5;12(5):298. doi: 10.3390/bios12050298.

Abstract

Hepatitis C virus (HCV) infections occur in approximately 3% of the world population. The development of an enhanced and extensive-scale screening is required to accomplish the World Health Organization's (WHO) goal of eliminating HCV as a public health problem by 2030. However, standard testing methods are time-consuming, expensive, and challenging to deploy in remote and underdeveloped areas. Therefore, a cost-effective, rapid, and accurate point-of-care (POC) diagnostic test is needed to properly manage the disease and reduce the economic burden caused by high case numbers. Herein, we present a fully automated reverse-transcription loop-mediated isothermal amplification (RT-LAMP)-based molecular diagnostic set-up for rapid HCV detection. The set-up consists of an automated disposable microfluidic chip, a small surface heater, and a reusable magnetic actuation platform. The microfluidic chip contains multiple chambers in which the plasma sample is processed. The system utilizes SYBR green dye to detect the amplification product with the naked eye. The efficiency of the microfluidic chip was tested with human plasma samples spiked with HCV virions, and the limit of detection observed was 500 virions/mL within 45 min. The entire virus detection process was executed inside a uniquely designed, inexpensive, disposable, and self-driven microfluidic chip with high sensitivity and specificity.

摘要

丙型肝炎病毒 (HCV) 感染约占世界人口的 3%。为了实现世界卫生组织 (WHO) 到 2030 年消除 HCV 作为公共卫生问题的目标,需要进行强化和大规模的筛查。然而,标准检测方法耗时、昂贵,并且在偏远和欠发达地区难以部署。因此,需要一种经济高效、快速且准确的即时护理 (POC) 诊断测试,以正确管理该疾病并减轻高病例数带来的经济负担。在此,我们提出了一种全自动基于逆转录环介导等温扩增 (RT-LAMP) 的分子诊断装置,用于快速检测 HCV。该装置由一个自动化的一次性微流控芯片、一个小型表面加热器和一个可重复使用的磁驱动平台组成。微流控芯片包含多个腔室,用于处理血浆样本。该系统利用 SYBR 绿染料通过肉眼检测扩增产物。我们用人血浆样本中添加 HCV 病毒颗粒对微流控芯片的效率进行了测试,在 45 分钟内观察到的检测限为 500 个病毒颗粒/mL。整个病毒检测过程在一个独特设计的、廉价的、一次性的、自驱动的微流控芯片内执行,具有高灵敏度和特异性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de1a/9138684/9118e8eb2682/biosensors-12-00298-g001.jpg

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