2023 Fiscal Year Annual Research Report
Lyophilization of RPA reagents for detection of SARS CoV-2 DNA and RNA
| Project/Area Number |
23KJ1201
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| Research Institution | Kyoto University |
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Principal Investigator |
Juma Kevin 京都大学, 農学研究科, 特別研究員(DC2)
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| Project Period (FY) |
2023-04-25 – 2024-03-31
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| Keywords | DNA amplification / Thermostable enzyme / Lyophilization / Pyruvate kinase |
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| Outline of Annual Research Achievements |
My research focused on establishing an isothermal recombinase polymerase amplification (RPA), a method conducted at around 41°C, utilizing recombinase, single-stranded DNA binding protein, strand-displacing DNA polymerase, and an ATP regenerating enzyme. I engineered a pyruvate kinase (Tma-PK) from thermophilic bacterium Thermotoga maritima as the ATP regenerating enzyme in RPA. I successfully optimized lyophilization conditions for RPA. The lyophilized RPA reagents remained stable for over 28 days at room temperature. RPA reagents containing Tma-PK exhibited superior performance to those with human PK suggesting that thermostable enzymes are preferable to mesophilic ones in RPA. I disseminated these findings in two domestic conferences and published two peer-reviewed articles.
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Research Products
(5 results)
