TY - JOUR
T1 - Rapid and simple detection of Ebola virus by reverse transcription-loop-mediated isothermal amplification
AU - Kurosaki, Yohei
AU - Takada, Ayato
AU - Ebihara, Hideki
AU - Grolla, Allen
AU - Kamo, Naoki
AU - Feldmann, Heinz
AU - Kawaoka, Yoshihiro
AU - Yasuda, Jiro
N1 - Funding Information:
We thank Toshie Sakuma and Aiko Fukuma for excellent technical assistance. This work was supported by grants from the Japan Science and Technology Agency (JST) and the Japan Society for the Promotion of Science (JSPS).
Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2007/4
Y1 - 2007/4
N2 - Ebola virus (EBOV) causes severe hemorrhagic fever in humans and nonhuman primates with high mortality rates. Rapid identification of the virus is required to prevent spread of the infection. In this study, we developed and evaluated a one-step simple reverse transcription-loop mediated isothermal amplification (RT-LAMP) assay for the rapid detection of Zaire ebolavirus (ZEBOV), the most virulent species of EBOV, targeting the trailer region of the viral genome. The assay could detect 20 copies of the artificial ZEBOV RNA in 26 min with a real time-monitoring detection, and also detect 10-3 FFU of the cell-culture propagated viruses. The reaction time needed to detect 104 FFU of ZEBOV was only 20 min. In addition, the assay was highly specific for ZEBOV. The RT-LAMP assay developed in this study is rapid, simple, highly specific, and sensitive for the detection of ZEBOV, and so may be an effective diagnostic tool. Furthermore, as this technique does not require sophisticated instrumentation, it seems very suitable for diagnosis in the field or laboratories in Ebola outbreak areas such as Central Africa.
AB - Ebola virus (EBOV) causes severe hemorrhagic fever in humans and nonhuman primates with high mortality rates. Rapid identification of the virus is required to prevent spread of the infection. In this study, we developed and evaluated a one-step simple reverse transcription-loop mediated isothermal amplification (RT-LAMP) assay for the rapid detection of Zaire ebolavirus (ZEBOV), the most virulent species of EBOV, targeting the trailer region of the viral genome. The assay could detect 20 copies of the artificial ZEBOV RNA in 26 min with a real time-monitoring detection, and also detect 10-3 FFU of the cell-culture propagated viruses. The reaction time needed to detect 104 FFU of ZEBOV was only 20 min. In addition, the assay was highly specific for ZEBOV. The RT-LAMP assay developed in this study is rapid, simple, highly specific, and sensitive for the detection of ZEBOV, and so may be an effective diagnostic tool. Furthermore, as this technique does not require sophisticated instrumentation, it seems very suitable for diagnosis in the field or laboratories in Ebola outbreak areas such as Central Africa.
KW - Detection
KW - Diagnosis
KW - Ebola virus
KW - Loop-mediated isothermal amplification
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U2 - 10.1016/j.jviromet.2006.11.031
DO - 10.1016/j.jviromet.2006.11.031
M3 - Article
C2 - 17194485
AN - SCOPUS:33847034418
SN - 0166-0934
VL - 141
SP - 78
EP - 83
JO - Journal of Virological Methods
JF - Journal of Virological Methods
IS - 1
ER -