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一种用于多种呼吸道病毒筛选的集成双层微流控平台。

An integrated dual-layer microfluidic platform for multiple respiratory viruses screening.

机构信息

Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.

Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.

出版信息

Anal Chim Acta. 2023 Feb 15;1242:340812. doi: 10.1016/j.aca.2023.340812. Epub 2023 Jan 7.

DOI:10.1016/j.aca.2023.340812
PMID:36657884
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9822946/
Abstract

Currently, the coronavirus disease 2019 (COVID-19) caused by the outbreak of a novel coronavirus (SARS-CoV-2) is spreading rapidly worldwide. Due to the high incidence of influenza coinciding with SARS-CoV-2, rapid detection is crucial to prevent spreading. Here, we present an integrated dual-layer microfluidic platform for specific and highly sensitive SARS-CoV-2, influenza viruses A (FluA) H1N1, H3N2, and influenza virus B (FluB) simultaneous detection. The platform includes a dual microchip (Dμchip) and a portable detection device for real-time fluorescence detection, temperature control and online communication. The Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) and Cas12a cleavage were performed on the Dμchip. The limit of detection (LoD) of the Dμchip assay was 10 copies for SARS-CoV-2, FluA H1N1, H3N2, and FluB RNAs. The Dμchip assay yielded no cross-reactivity against other coronaviruses, so it was suitable for the screening of multiple viruses. Moreover, the positive percentage agreement (PPA) and negative percentage agreement (NPA) of the assay were 97.9% and 100%, respectively, in 75 clinical samples compared to data from RT-PCR-based assays. Furthermore, the assay allowed the detection SARS-CoV-2 and influenza viruses in spiked samples. Overall, the present platform would provide a rapid method for the screening of multiple viruses in hospital emergency, community and primary care settings and facilitate the remote diagnosis and outbreak control of the COVID-19.

摘要

目前，由新型冠状病毒（SARS-CoV-2）引发的 2019 年冠状病毒病（COVID-19）在全球范围内迅速传播。由于流感与 SARS-CoV-2 的高发率，快速检测对于防止传播至关重要。在这里，我们提出了一种用于特异性和高灵敏度 SARS-CoV-2、甲型流感病毒（FluA）H1N1、H3N2 和乙型流感病毒（FluB）同时检测的集成双层微流控平台。该平台包括一个双层微芯片（Dμchip）和一个便携式检测设备，用于实时荧光检测、温度控制和在线通信。逆转录环介导等温扩增（RT-LAMP）和 Cas12a 切割在 Dμchip 上进行。Dμchip 测定的检测限（LoD）为 SARS-CoV-2、FluA H1N1、H3N2 和 FluB RNA 的 10 个拷贝。Dμchip 测定法对其他冠状病毒没有交叉反应，因此适用于多种病毒的筛选。此外，该测定法在与基于 RT-PCR 的测定法相比，在 75 个临床样本中的阳性百分比一致率（PPA）和阴性百分比一致率（NPA）分别为 97.9%和 100%。此外，该测定法允许在添加样本中检测 SARS-CoV-2 和流感病毒。总的来说，该平台将为医院急诊、社区和初级保健环境中的多种病毒的筛选提供一种快速方法，并有助于 COVID-19 的远程诊断和疫情控制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/c9b0c7a81535/gr7_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/22a94f3298dd/ga1_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/e0b226ba78cf/gr1_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/399640db1e53/gr2_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/475807506709/gr3_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/6ce4d46cab14/gr4_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/4b32b2de9b1c/gr5_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/1313d1a1e4b9/gr6_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/c9b0c7a81535/gr7_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/22a94f3298dd/ga1_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/e0b226ba78cf/gr1_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/399640db1e53/gr2_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/475807506709/gr3_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/6ce4d46cab14/gr4_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/4b32b2de9b1c/gr5_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/1313d1a1e4b9/gr6_lrg.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2be/9822946/c9b0c7a81535/gr7_lrg.jpg

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