Research Abstract

等温増幅および携行可能な塩基配列解読装置を用いたデングウイルスの血清型判定 ブックマーク

Serotyping dengue virus with isothermal amplification and a portable sequencer

2017年6月14日 Scientific Reports 7 : 3510 doi: 10.1038/s41598-017-03734-5 (2017)


Junya Yamagishi, Lucky R. Runtuwene, Kyoko Hayashida, Arthur E. Mongan, Lan Anh Nguyen Thi, Linh Nguyen Thuy, Cam Nguyen Nhat, Kriengsak Limkittikul, Chukiat Sirivichayakul, Nuankanya Sathirapongsasuti, Martin Frith, Wojciech Makalowski, Yuki Eshita, Sumio Sugano and Yutaka Suzuki

Corresponding Author

鈴木 穣
東京大学大学院 新領域創成科学研究科 メディカル情報生命専攻

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.