@article{fc8c0fb66ce64a5884a4e306f0a5a939,
title = "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-assisted and Coupled Amplification) reaction, that consists of a simple one-pot format of {\textquoteleft}sample-in and result-out{\textquoteright} 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.",
author = "Radhika Biyani and Kirti Sharma and Kenji Kojima and Madhu Biyani and Vishnu Sharma and Tarun Kumawat and Juma, {Kevin Maafu} and Itaru Yanagihara and Shinsuke Fujiwara and Eiichi Kodama and Yuzuru Takamura and Masahiro Takagi and Kiyoshi Yasukawa and Manish Biyani",
note = "Funding Information: This work was supported in part by Grant-in-aid funding for the COVID-19 consortium (Grant no. BT/ COVID0020/01/20 to M.B.) from the Biotechnology Industry Research Assistance Council, a Government of India enterprise; Emerging/re-emerging infectious disease project of Japan (Grant no. JP20he0622020 to K.K., I.Y., S.F., E.K., K.Y., and M.B. and Grant no. JP20fk0108143 to I.Y., S.F., and K.Y.) from the Japan Agency for Medical Research and Development; and Adaptable and Seamless Technology Transfer Program through Target-driven R&D (A-STEP, No. JPMJTR20UU to K.Y. and Y.T.) from the Japan Science and Technology Agency (JST) of Japan. We thank Ms. Ruchika Mishra of Tsukahara Laboratory of the Japan Advanced Institute of Science and Technology and Ms. Keiko Kawai of BioSeeds Corporation for supporting the E. coli cell-related experiments. Publisher Copyright: {\textcopyright} 2021, The Author(s).",
year = "2021",
month = dec,
doi = "10.1038/s41598-021-95411-x",
language = "English",
volume = "11",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",
number = "1",
}