Multiplexed detection of respiratory pathogens with a portable analyzer in a “raw-sample-in and answer-out” manner

Abstract Coronavirus disease 2019 (COVID-19) has emerged, rapidly spread and caused significant morbidity and mortality worldwide. There is an urgent public health need for rapid, sensitive, specific, and on-site diagnostic tests for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infec...

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Autores principales:	Nan Li, Minjie Shen, Jiajia Liu, Li Zhang, Huili Wang, Youchun Xu, Jing Cheng
Formato:	article
Lenguaje:	EN
Publicado:	Nature Publishing Group 2021
Materias:	Technology T Engineering (General). Civil engineering (General) TA1-2040
Acceso en línea:	https://doaj.org/article/25f0ce16bc89460f9bb0f2ae40bbfa80
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spelling	oai:doaj.org-article:25f0ce16bc89460f9bb0f2ae40bbfa802021-11-28T12:08:00ZMultiplexed detection of respiratory pathogens with a portable analyzer in a “raw-sample-in and answer-out” manner10.1038/s41378-021-00321-72055-7434https://doaj.org/article/25f0ce16bc89460f9bb0f2ae40bbfa802021-11-01T00:00:00Zhttps://doi.org/10.1038/s41378-021-00321-7https://doaj.org/toc/2055-7434Abstract Coronavirus disease 2019 (COVID-19) has emerged, rapidly spread and caused significant morbidity and mortality worldwide. There is an urgent public health need for rapid, sensitive, specific, and on-site diagnostic tests for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. In this study, a fully integrated and portable analyzer was developed to detect SARS-CoV-2 from swab samples based on solid-phase nucleic acid extraction and reverse transcription loop-mediated isothermal amplification (RT-LAMP). The swab can be directly inserted into a cassette for multiplexed detection of respiratory pathogens without pre-preparation. The overall detection process, including swab rinsing, magnetic bead-based nucleic acid extraction, and 8-plex real-time RT-LAMP, can be automatically performed in the cassette within 80 min. The functionality of the cassette was validated by detecting the presence of a SARS-CoV-2 pseudovirus and three other respiratory pathogens, i.e., Klebsiella pneumoniae, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. The limit of detection (LoD) for the SARS-CoV-2 pseudovirus was 2.5 copies/μL with both primer sets (N gene and ORF1ab gene), and the three bacterial species were successfully detected with an LoD of 2.5 colony-forming units (CFU)/μL in 800 μL of swab rinse. Thus, the analyzer developed in this study has the potential to rapidly detect SARS-CoV-2 and other respiratory pathogens on site in a “raw-sample-in and answer-out” manner.Nan LiMinjie ShenJiajia LiuLi ZhangHuili WangYouchun XuJing ChengNature Publishing GrouparticleTechnologyTEngineering (General). Civil engineering (General)TA1-2040ENMicrosystems & Nanoengineering, Vol 7, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Technology T Engineering (General). Civil engineering (General) TA1-2040
spellingShingle	Technology T Engineering (General). Civil engineering (General) TA1-2040 Nan Li Minjie Shen Jiajia Liu Li Zhang Huili Wang Youchun Xu Jing Cheng Multiplexed detection of respiratory pathogens with a portable analyzer in a “raw-sample-in and answer-out” manner
description	Abstract Coronavirus disease 2019 (COVID-19) has emerged, rapidly spread and caused significant morbidity and mortality worldwide. There is an urgent public health need for rapid, sensitive, specific, and on-site diagnostic tests for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. In this study, a fully integrated and portable analyzer was developed to detect SARS-CoV-2 from swab samples based on solid-phase nucleic acid extraction and reverse transcription loop-mediated isothermal amplification (RT-LAMP). The swab can be directly inserted into a cassette for multiplexed detection of respiratory pathogens without pre-preparation. The overall detection process, including swab rinsing, magnetic bead-based nucleic acid extraction, and 8-plex real-time RT-LAMP, can be automatically performed in the cassette within 80 min. The functionality of the cassette was validated by detecting the presence of a SARS-CoV-2 pseudovirus and three other respiratory pathogens, i.e., Klebsiella pneumoniae, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. The limit of detection (LoD) for the SARS-CoV-2 pseudovirus was 2.5 copies/μL with both primer sets (N gene and ORF1ab gene), and the three bacterial species were successfully detected with an LoD of 2.5 colony-forming units (CFU)/μL in 800 μL of swab rinse. Thus, the analyzer developed in this study has the potential to rapidly detect SARS-CoV-2 and other respiratory pathogens on site in a “raw-sample-in and answer-out” manner.
format	article
author	Nan Li Minjie Shen Jiajia Liu Li Zhang Huili Wang Youchun Xu Jing Cheng
author_facet	Nan Li Minjie Shen Jiajia Liu Li Zhang Huili Wang Youchun Xu Jing Cheng
author_sort	Nan Li
title	Multiplexed detection of respiratory pathogens with a portable analyzer in a “raw-sample-in and answer-out” manner
title_short	Multiplexed detection of respiratory pathogens with a portable analyzer in a “raw-sample-in and answer-out” manner
title_full	Multiplexed detection of respiratory pathogens with a portable analyzer in a “raw-sample-in and answer-out” manner
title_fullStr	Multiplexed detection of respiratory pathogens with a portable analyzer in a “raw-sample-in and answer-out” manner
title_full_unstemmed	Multiplexed detection of respiratory pathogens with a portable analyzer in a “raw-sample-in and answer-out” manner
title_sort	multiplexed detection of respiratory pathogens with a portable analyzer in a “raw-sample-in and answer-out” manner
publisher	Nature Publishing Group
publishDate	2021
url	https://doaj.org/article/25f0ce16bc89460f9bb0f2ae40bbfa80
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Multiplexed detection of respiratory pathogens with a portable analyzer in a “raw-sample-in and answer-out” manner

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