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タイトル: | Development of a point-of-care test to detect SARS-CoV-2 from saliva which combines a simple RNA extraction method with colorimetric reverse transcription loop-mediated isothermal amplification detection |
著者: | Yamazaki, Wataru https://orcid.org/0000-0002-7874-8014 (unconfirmed) Matsumura, Yasufumi https://orcid.org/0000-0001-8595-8944 (unconfirmed) Thongchankaew-Seo, Uraiwan Yamazaki, Yasuko Nagao, Miki https://orcid.org/0000-0002-8886-6145 (unconfirmed) |
著者名の別形: | 山崎, 渉 松村, 康史 山崎, 安子 長尾, 美紀 |
キーワード: | COVID-19 LAMP Point-of-care test POCT SARS-CoV-2 semi alkaline proteinase |
発行日: | Mar-2021 |
出版者: | Elsevier BV |
誌名: | Journal of Clinical Virology |
巻: | 136 |
論文番号: | 104760 |
抄録: | The new coronavirus infection (COVID-19) is a major public health concern, with a high burden and risk for infection among patients and healthcare workers. Saliva droplets containing SARS-COV-2 are a major vector for COVID-19 infection, making saliva a promising alternative for COVID-19 testing using nasopharyngeal swab samples. To diagnose COVID-19 patients in the field, a point-of-care test (POCT) using saliva was conceptualized. We have developed a simple method for extracting RNA from saliva samples using semi-alkaline proteinase, a sputum homogenizer typically used for preparing samples for tuberculosis testing, and a subsequent simple heating step with no need for centrifugation or RNA extraction. Further, we newly developed a triplex reverse transcription loop-mediated isothermal amplification approach (RT-LAMP) which utilizes colorimetric readout using a heat block, with results evaluated with the unaided eye. In 44 clinical patients suspected of having COVID-19 infection, the test took 45 min, and resulted in a diagnostic sensitivity of 82.6% (19/23) and diagnostic specificity of 100% (21/21), compared to the reference standard. The limit of detection was 250 copies/reaction (25, 000 copies/mL). Our newly developed POCT approach achieved simple RNA extraction and constant RT-LAMP detection. This POCT has the potential to be used for simple inspection stations in a field setting, helping reduce the risk of infection by simplifying and accelerating testing for COVID-19. |
著作権等: | © 2021 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY license |
URI: | http://hdl.handle.net/2433/263826 |
DOI(出版社版): | 10.1016/j.jcv.2021.104760 |
PubMed ID: | 33610926 |
出現コレクション: | 学術雑誌掲載論文等 |
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