Rapid and simple detection of Ebola virus by reverse transcription-loop-mediated isothermal amplification.
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 10(4) 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.
Kurosaki
Yohei
Y
National Research Institute of Police Science, Kashiwa 277-0882, Japan.
Takada
Ayato
A
Ebihara
Hideki
H
Grolla
Allen
A
Kamo
Naoki
N
Feldmann
Heinz
H
Kawaoka
Yoshihiro
Y
Yasuda
Jiro
J
D000818:Animals; D002522:Chlorocebus aethiops; D029043:Ebolavirus / Q000235:genetics / Q000302:isolation & purification*; D005069:Evaluation Studies as Topic; D019142:Hemorrhagic Fever, Ebola / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014709:Vero Cells
D012367:RNA, Viral