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Serotyping dengue virus with isothermal amplification and a portable sequencer

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Title: Serotyping dengue virus with isothermal amplification and a portable sequencer
Authors: Yamagishi, Junya Browse this author →KAKEN DB
Runtuwene, Lucky R. Browse this author
Hayashida, Kyoko Browse this author
Mongan, Arthur E. Browse this author
Lan Anh Nguyen Thi Browse this author
Linh Nguyen Thuy Browse this author
Cam Nguyen Nhat Browse this author
Limkittikul, Kriengsak Browse this author
Sirivichayakul, Chukiat Browse this author
Sathirapongsasuti, Nuankanya Browse this author
Frith, Martin Browse this author
Makalowski, Wojciech Browse this author
Eshita, Yuki Browse this author
Sugano, Sumio Browse this author
Suzuki, Yutaka Browse this author
Issue Date: 14-Jun-2017
Publisher: Nature Publishing Group
Journal Title: Scientific reports
Volume: 7
Start Page: 3510
Publisher DOI: 10.1038/s41598-017-03734-5
Abstract: The recent development of a nanopore-type portable DNA sequencer has changed the way we think about DNA sequencing. We can perform sequencing directly in the field, where we collect the samples. Here, we report the development of a novel method to detect and genotype tropical disease pathogens, using dengue fever as a model. By combining the sequencer with isothermal amplification that only requires a water bath, we were able to amplify and sequence target viral genomes with ease. Starting from a serum sample, the entire procedure could be finished in a single day. The analysis of blood samples collected from 141 Indonesian patients demonstrated that this method enables the clinical identification and serotyping of the dengue virus with high sensitivity and specificity. The overall successful detection rate was 79%, and a total of 58 SNVs were detected. Similar analyses were conducted on 80 Vietnamese and 12 Thai samples with similar performance. Based on the obtained sequence information, we demonstrated that this approach is able to produce indispensable information for etiologically analyzing annual or regional diversifications of the pathogens.
Type: article
Appears in Collections:人獣共通感染症リサーチセンター (Research Center for Zoonosis Control) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 山岸 潤也

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