Molecular diagnostics for clinical respiratory virus on a total integrated centrifugal microsystem using reverse transcription-loop-mediated isothermal amplification

被引：10

作者：

Phan V.M. ^{[1
]}

Kang S.W. ^{[2
]}

Kim Y.H. ^{[2
]}

Lee M.Y. ^{[3
]}

Nguyen H.V. ^{[1
]}

Jeon Y.L. ^{[4
]}

Lee W.I. ^{[3
]}

Seo T.S. ^{[1
]}

机构：

[1] Department of Chemical Engineering (BK21 FOUR Integrated Engineering Program), Kyung Hee University, Yongin

[2] Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Kyung Hee University Hospital at Gangdong, College of Medicine, Kyung Hee University, Seoul

[3] Department of Laboratory Medicine, Kyung Hee University College of Medicine, Kyung Hee University Hospital at Gangdong, Seoul

[4] Department of Laboratory Medicine, Green Cross Laboratories, Yongin-si

来源：

Sensors and Actuators B: Chemical | 2023年 / 390卷

基金：

新加坡国家研究基金会;

关键词：

Centrifugal microfluidics; COVID-19; Loop-mediated isothermal amplification; Point-of-care; Respiratory virus;

D O I：

10.1016/j.snb.2023.133962

中图分类号：

学科分类号：

摘要：

Respiratory viruses are highly contagious agents that can cause endemic and epidemic infections in humans. Early detection of these viruses is crucial in preventing economic damage and reducing mortality rates. In this study, we present a total integrated genetic analyzer to perform a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for the simultaneous detection of 7 respiratory viruses (Influenza A H1N1 and H3N2, Influenza B, Respiratory syncytial virus A and B, Adenovirus, and COVID-19). The primer sets for the RT-LAMP assay were designed and evaluated in comparison with the RT-PCR assay using clinical samples, confirming high specificity and efficiency. The entire process of viral RNA extraction, reagent mixing, gene amplification, and detection was completed on the device in 1 hr 20 min. The constructed portable diagnostic instrument is equipped with a rotary motor, two sets of peltier heaters, a fluorescence detector, and a touch screen for inputting experimental parameters and displaying result. The proposed point-of-care (POC) diagnostic platform correctly analyzed a total of 21 clinical samples (3 for each of the 7 viruses). The limit-of-detection (LOD) for Influenza A subtype H3N2 was 101 pfu/mL, which demonstrates the high performance of our proposed centrifugal microsystem for on-site molecular diagnostics in medical centers. © 2023 Elsevier B.V.

引用

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