Development of robust isothermal RNA amplification assay for lab-free testing of RNA viruses

Abstract Simple tests of infectiousness that return results in minutes and directly from samples even with low viral loads could be a potential game-changer in the fight against COVID-19. Here, we describe an improved isothermal nucleic acid amplification assay, termed the RICCA (RNA Isothermal Co-a...

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Autores principales:	Radhika Biyani, Kirti Sharma, Kenji Kojima, Madhu Biyani, Vishnu Sharma, Tarun Kumawat, Kevin Maafu Juma, Itaru Yanagihara, Shinsuke Fujiwara, Eiichi Kodama, Yuzuru Takamura, Masahiro Takagi, Kiyoshi Yasukawa, Manish Biyani
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/bcb2e1192fba4d1c9c295200b180c00e
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spelling	oai:doaj.org-article:bcb2e1192fba4d1c9c295200b180c00e2021-12-02T14:53:49ZDevelopment of robust isothermal RNA amplification assay for lab-free testing of RNA viruses10.1038/s41598-021-95411-x2045-2322https://doaj.org/article/bcb2e1192fba4d1c9c295200b180c00e2021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-95411-xhttps://doaj.org/toc/2045-2322Abstract Simple tests of infectiousness that return results in minutes and directly from samples even with low viral loads could be a potential game-changer in the fight against COVID-19. Here, we describe an improved isothermal nucleic acid amplification assay, termed the RICCA (RNA Isothermal Co-assisted and Coupled Amplification) reaction, that consists of a simple one-pot format of ‘sample-in and result-out’ with a primary focus on the detection of low copy numbers of RNA virus directly from saliva without the need for laboratory processing. We demonstrate our assay by detecting 16S rRNA directly from E. coli cells with a sensitivity as low as 8 CFU/μL and RNA fragments from a synthetic template of SARS-CoV-2 with a sensitivity as low as 1740 copies/μL. We further demonstrate the applicability of our assay for real-time testing at the point of care by designing a closed format for paper-based lateral flow assay and detecting heat-inactivated SARS-COV-2 virus in human saliva at concentrations ranging from 28,000 to 2.8 copies/μL with a total assay time of 15–30 min.Radhika BiyaniKirti SharmaKenji KojimaMadhu BiyaniVishnu SharmaTarun KumawatKevin Maafu JumaItaru YanagiharaShinsuke FujiwaraEiichi KodamaYuzuru TakamuraMasahiro TakagiKiyoshi YasukawaManish BiyaniNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Radhika Biyani Kirti Sharma Kenji Kojima Madhu Biyani Vishnu Sharma Tarun Kumawat Kevin Maafu Juma Itaru Yanagihara Shinsuke Fujiwara Eiichi Kodama Yuzuru Takamura Masahiro Takagi Kiyoshi Yasukawa Manish Biyani Development of robust isothermal RNA amplification assay for lab-free testing of RNA viruses
description	Abstract Simple tests of infectiousness that return results in minutes and directly from samples even with low viral loads could be a potential game-changer in the fight against COVID-19. Here, we describe an improved isothermal nucleic acid amplification assay, termed the RICCA (RNA Isothermal Co-assisted and Coupled Amplification) reaction, that consists of a simple one-pot format of ‘sample-in and result-out’ with a primary focus on the detection of low copy numbers of RNA virus directly from saliva without the need for laboratory processing. We demonstrate our assay by detecting 16S rRNA directly from E. coli cells with a sensitivity as low as 8 CFU/μL and RNA fragments from a synthetic template of SARS-CoV-2 with a sensitivity as low as 1740 copies/μL. We further demonstrate the applicability of our assay for real-time testing at the point of care by designing a closed format for paper-based lateral flow assay and detecting heat-inactivated SARS-COV-2 virus in human saliva at concentrations ranging from 28,000 to 2.8 copies/μL with a total assay time of 15–30 min.
format	article
author	Radhika Biyani Kirti Sharma Kenji Kojima Madhu Biyani Vishnu Sharma Tarun Kumawat Kevin Maafu Juma Itaru Yanagihara Shinsuke Fujiwara Eiichi Kodama Yuzuru Takamura Masahiro Takagi Kiyoshi Yasukawa Manish Biyani
author_facet	Radhika Biyani Kirti Sharma Kenji Kojima Madhu Biyani Vishnu Sharma Tarun Kumawat Kevin Maafu Juma Itaru Yanagihara Shinsuke Fujiwara Eiichi Kodama Yuzuru Takamura Masahiro Takagi Kiyoshi Yasukawa Manish Biyani
author_sort	Radhika Biyani
title	Development of robust isothermal RNA amplification assay for lab-free testing of RNA viruses
title_short	Development of robust isothermal RNA amplification assay for lab-free testing of RNA viruses
title_full	Development of robust isothermal RNA amplification assay for lab-free testing of RNA viruses
title_fullStr	Development of robust isothermal RNA amplification assay for lab-free testing of RNA viruses
title_full_unstemmed	Development of robust isothermal RNA amplification assay for lab-free testing of RNA viruses
title_sort	development of robust isothermal rna amplification assay for lab-free testing of rna viruses
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/bcb2e1192fba4d1c9c295200b180c00e
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Development of robust isothermal RNA amplification assay for lab-free testing of RNA viruses

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