Development of a Multiplex Loop-Mediated Isothermal Amplification (LAMP) Method for Simultaneous Detection of Spotted Fever Group Rickettsiae and Malaria Parasites by Dipstick DNA Chromatography

Moonga, Lavel Chinyama; Hayashida, Kyoko; Kawai, Naoko et al.

Diagnostics, 2020, 10(11), 897

Permalink : https://hdl.handle.net/2115/80170

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アクセス権
open access
URI
https://hdl.handle.net/2115/80170
タイトル
Development of a Multiplex Loop-Mediated Isothermal Amplification (LAMP) Method for Simultaneous Detection of Spotted Fever Group Rickettsiae and Malaria Parasites by Dipstick DNA Chromatography
著者
Moonga, Lavel Chinyama ; Hayashida, Kyoko ; Kawai, Naoko ; Nakao, Ryo ; Sugimoto, Chihiro ; Namangala, Boniface ; Yamagishi, Junya
著者名
Moonga, Lavel Chinyama
著者名
Hayashida, Kyoko
著者名
Kawai, Naoko
著者名
Nakao, Ryo
著者名
Sugimoto, Chihiro
著者名
Namangala, Boniface
著者名
Yamagishi, Junya
言語
英語
キーワード
spotted fever group rickettsia
SFG rickettsiosis
malaria
multiplex detection
loop-mediated isothermal amplification (LAMP)
dipstick DNA chromatography
発行日
2020-11
出版者
MDPI
収録物名
Diagnostics
巻(号)
10(11)
ページ
897
抄録
Spotted fever group (SFG) rickettsiae causes febrile illness in humans worldwide. Since SFG rickettsiosis's clinical presentation is nonspecific, it is frequently misdiagnosed as other febrile diseases, especially malaria, and complicates proper treatment. Aiming at rapid, simple, and simultaneous detection of SFG Rickettsia spp. and Plasmodium spp., we developed a novel multiple pathogen detection system by combining a loop-mediated isothermal amplification (LAMP) method and dipstick DNA chromatography technology. Two primer sets detecting SFG Rickettsia spp. and Plasmodium spp. were mixed, and amplified products were visualized by hybridizing to dipstick DNA chromatography. The multiplex LAMP with dipstick DNA chromatography distinguished amplified Rickettsia and Plasmodium targeted genes simultaneously. The determined sensitivity using synthetic nucleotides was 1000 copies per reaction for mixed Rickettsia and Plasmodium genes. When genomic DNA from in vitro cultured organisms was used, the sensitivity was 100 and 10 genome equivalents per reaction for Rickettsia monacensis and Plasmodium falciparum, respectively. Although further improvement will be required for more sensitive detection, our developed simultaneous diagnosis technique will contribute to the differential diagnosis of undifferentiated febrile illness caused by either SFG Rickettsia spp. or Plasmodium spp. in resource-limited endemic areas. Importantly, this scheme is potentially versatile for the simultaneous detection of diverse infectious diseases.
資源タイプ
学術雑誌論文
出版タイプ
Version of Record
権利情報
Creative Commons Attribution 4.0 International
関連情報 (isIdenticalTo)
出版社版DOI
https://doi.org/10.3390/diagnostics10110897