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Development of robust isothermal RNA amplification assay for lab-free testing of RNA viruses.
Biyani, Radhika; Sharma, Kirti; Kojima, Kenji; Biyani, Madhu; Sharma, Vishnu; Kumawat, Tarun; Juma, Kevin Maafu; Yanagihara, Itaru; Fujiwara, Shinsuke; Kodama, Eiichi; Takamura, Yuzuru; Takagi, Masahiro; Yasukawa, Kiyoshi; Biyani, Manish.
  • Biyani R; Department of Bioscience and Biotechnology, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi City, Ishikawa, 923-1292, Japan.
  • Sharma K; BioSeeds Corporation, JAIST Venture Business Laboratory, Ishikawa Create Labo, Asahidai 2-13, Nomi City, Ishikawa, 923-1211, Japan.
  • Kojima K; Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan.
  • Biyani M; BioSeeds Corporation, JAIST Venture Business Laboratory, Ishikawa Create Labo, Asahidai 2-13, Nomi City, Ishikawa, 923-1211, Japan.
  • Sharma V; Biyani BioSolutions Pvt. Ltd., Biyani Group of Colleges Venture Business Laboratory, R-4, Sector 3, Vidhyadhar Nagar, Jaipur, 302039, India.
  • Kumawat T; Biyani BioSolutions Pvt. Ltd., Biyani Group of Colleges Venture Business Laboratory, R-4, Sector 3, Vidhyadhar Nagar, Jaipur, 302039, India.
  • Juma KM; Biyani BioSolutions Pvt. Ltd., Biyani Group of Colleges Venture Business Laboratory, R-4, Sector 3, Vidhyadhar Nagar, Jaipur, 302039, India.
  • Yanagihara I; Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan.
  • Fujiwara S; Department of Developmental Medicine, Research Institute, Osaka Women's and Children's Hospital, 840 Murodocho, Izumi, Osaka, 594-1101, Japan.
  • Kodama E; Department of Biosciences, School of Biological and Environmental Sciences, Kwansei-Gakuin University, 2-1 Gakuen, Sanda, Hyogo, 669-1337, Japan.
  • Takamura Y; Division of Infectious Diseases, International Research Institute of Disaster Science, Tohoku University, 2-1 Seiryocho Aoba-ku, Sendai, Miyagi, 980-8575, Japan.
  • Takagi M; Department of Bioscience and Biotechnology, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi City, Ishikawa, 923-1292, Japan.
  • Yasukawa K; Department of Bioscience and Biotechnology, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi City, Ishikawa, 923-1292, Japan.
  • Biyani M; Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan.
Sci Rep ; 11(1): 15997, 2021 08 06.
Article in English | MEDLINE | ID: covidwho-1345579
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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA Viruses / RNA, Viral / Point-of-Care Systems / Nucleic Acid Amplification Techniques / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study Limits: Humans Language: English Journal: Sci Rep Year: 2021 Document Type: Article Affiliation country: S41598-021-95411-x

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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA Viruses / RNA, Viral / Point-of-Care Systems / Nucleic Acid Amplification Techniques / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study Limits: Humans Language: English Journal: Sci Rep Year: 2021 Document Type: Article Affiliation country: S41598-021-95411-x