We developed an original nucleic acid amplification technology, “Smart-15* The development was performed in collaboration with the RIKEN Preventive Medicine & Diag-nosis Innovation Program (PMI), Kanagawa Prefecture and K.K. DNAFORM.Amp”, which is an isothermal rapid DNA amplification technology. In 2020, the worldwide pandemic caused by the novel coronavirus, SARS-CoV-2 resulted in a dramatic change in our lifestyle. To contribute to resolving this once-in-a-century epidemic situation, we concentrated almost all of our efforts in this year to develop a SARS-CoV-2 rapid detection kit* using the SmartAmp method.On 17 January 2020, the first novel SARS-CoV-2 genome sequence (strain Wuhan-Hu-1, NC_045512.2) was deposited in NCBI/GenBank, and we im-mediately began to construct a SARS-CoV-2-specific SmartAmp primer set by selecting the Orf1b-Nsp15 region as a target of the primers. We evaluated the primer set with authentic genomic RNA isolated from a passenger on the Diamond Princess cruise ship (provided by the Kanagawa Prefectural Institute of Public Health). Ultimately, our newly-developed SmartAmp SARS-CoV-2 detection kit was approved as a IVD (in vitro diagnosis) kit on 17 August 2020. The detection kit is shipped from a collaborating company “K.K. DNAFORM”. The kit was used not only at hospitals in Kanagawa, but also by several Japanese sport federations and associations (All Japan Judo Federation, Japan Profession-al Bowling Association, and Japan Football Association, etc.) for SARS-CoV-2 testing.The SARS-CoV-2 detection kit can be used with our pretreatment method, “SmartExtract”, which is able to isolate viral RNA from nasopharyngeal swabs or saliva. Currently, a collaborating company has provided several devices based on our systems that allow for the initial RNA isolation all the way to identification of the virus. Thus, the convenience of the testing kit has been im-proved considerably. Additionally, we have developed a prototype of a “Direct-SmartAmp method”, which does not require the RNA-isolation step, i.e., the sample can be directly mixed in the testing reagent.Figure: SmartAmp SARS-CoV-2 detection kitA, Reagents of the detection kit. The kit consists of only two reagents, Reagent E (left tube, includes enzyme, dNTP and reaction buffer) and Reagent P (right tube, includes primer mix with fluorescence oligonucleotide, Eprimer™).B, Amplification results using the detection kit. The SARS-CoV-2 virus in the isolated sample is rapidly identified, within 30 min, by increasing fluorescence intensity.Recent Major PublicationsSoma T and Usui K. Nucleic acid amplification test for SARS-CoV-2 in Japan. Clin Immunol & Allergol 75, 101-108 (2021)Hanami T, Tanabe T, Hanashi T, Yamaguchi M, Nakata H, Mitani Y, Kimura Y, Soma T, Usui K, Isobe M, Ogawa T, Itoh M, Hayashizaki Y and Kondo S. Scanning single-molecule counting system for Eprobe with highly simple and effective approach. Plos One 15, e0243319 (2020)Tokumoto S, Miyata Y, Usui K, Deviatiiarov R, Ohkawa T, Kondratieva S, Shagimardanova E, Gusev O, Cornette R, Itoh M, Hayashizaki Y and Kikawada T. Development of a Tet-On Inducible Expression System for the Anhydrobi-otic Cell Line, Pv11. Insects 11, 781 (2020)Genetic Diagnosis Technology UnitNucleic Acid Diagnosis System Developing UnitUnit Leader: Kengo Usui
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