title: Survey and Visual Detection of Zaire ebolavirus in Clinical Samples Targeting the Nucleoprotein Gene in Sierra Leone.
abstract: Ebola virus (EBOV) can lead to severe hemorrhagic fever with a high risk of death in humans and other primates. To guide treatment and prevent spread of the viral infection, a rapid and sensitive detection method is required for clinical samples. Here, we described and evaluated a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method to detect Zaire ebolavirus using the nucleoprotein gene (NP) as a target sequence. Two different techniques were used, a calcein/Mn(2+) complex chromogenic method and real-time turbidity monitoring. The RT-LAMP assay detected the NP target sequence with a limit of 4.56 copies/μL within 45 min under 61°C, a similar even or increase in sensitivity than that of real-time reverse transcription-polymerase chain reaction (RT-PCR). Additionally, all pseudoviral particles or non- Zaire EBOV genomes were negative for LAMP detection, indicating that the assay was highly specific for EBOV. To appraise the availability of the RT-LAMP method for use in clinical diagnosis of EBOV, of 417 blood or swab samples collected from patients with clinically suspected infections in Sierra Leone, 307 were identified for RT-LAMP-based surveillance of EBOV. Therefore, the highly specific and sensitive RT-LAMP method allows the rapid detection of EBOV, and is a suitable tool for clinical screening, diagnosis, and primary quarantine purposes.
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.
authors.forename: Huan
authors.initials: H
authors.lastname: Li
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.
authors.forename: Xuesong
authors.initials: X
authors.lastname: Wang
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Wei
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.
authors.forename: Weishi
authors.initials: W
authors.lastname: Lin
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.
authors.forename: Erna
authors.initials: E
authors.lastname: Li
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.
authors.forename: Puyuan
authors.initials: P
authors.lastname: Li
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.
authors.forename: Derong
authors.initials: D
authors.lastname: Dong
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.
authors.forename: Lifei
authors.initials: L
authors.lastname: Cui
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.
authors.forename: Xuan
authors.initials: X
authors.lastname: Hu
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.
authors.forename: Boxing
authors.initials: B
authors.lastname: Li
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.
authors.forename: Xiangna
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Liu
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Yuan
keywords: RT-LAMP; Zaire EBOV; prevalence; rapid detection; sensitivity; specificity