title: Development and Evaluation of Reverse Transcription-Loop-Mediated Isothermal Amplification (RT-LAMP) Assay Coupled with a Portable Device for Rapid Diagnosis of Ebola Virus Disease in Guinea.
abstract: Given the current absence of specific drugs or vaccines for Ebola virus disease (EVD), rapid, sensitive, and reliable diagnostic methods are required to stem the transmission chain of the disease. We have developed a rapid detection assay for Zaire ebolavirus based on reverse transcription-loop-mediated isothermal amplification (RT-LAMP) and coupled with a novel portable isothermal amplification and detection platform. The RT-LAMP assay is based on primer sets that target the untranscribed trailer region or nucleoprotein coding region of the viral RNA. The test could specifically detect viral RNAs of Central and West African Ebola virus strains within 15 minutes with no cross-reactivity to other hemorrhagic fever viruses and arboviruses, which cause febrile disease. The assay was evaluated using a total of 100 clinical specimens (serum, n = 44; oral swab, n = 56) collected from suspected EVD cases in Guinea. The specificity of this diagnostic test was 100% for both primer sets, while the sensitivity was 100% and 97.9% for the trailer and nucleoprotein primer sets, respectively, compared with a reference standard RT-PCR test. These observations suggest that our diagnostic assay is useful for identifying EVD cases, especially in the field or in settings with insufficient infrastructure.
authors.affiliation: Institute of Tropical Medicine (NEKKEN), Nagasaki University, Nagasaki, Japan.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Kurosaki
authors.affiliation: Laboratoire des Fièvres Hémorragiques en Guinée, Conakry, Guinea.|Hôpital National Donka, Service des Maladies Infectieuses et Tropicales, Conakry, Guinea.|Université Gamal Abdel Nasser, Conakry, Guinea.
authors.forename: N'Faly
authors.initials: N
authors.lastname: Magassouba
authors.affiliation: Institute of Tropical Medicine (NEKKEN), Nagasaki University, Nagasaki, Japan.|Graduate School of Biomedical Sciences and Program for Nurturing Global Leaders in Tropical and Emerging Communicable Diseases, Nagasaki University, Nagasaki, Japan.
authors.forename: Olamide K
authors.initials: OK
authors.lastname: Oloniniyi
authors.affiliation: Institute of Tropical Medicine (NEKKEN), Nagasaki University, Nagasaki, Japan.
authors.forename: Mahamoud S
authors.initials: MS
authors.lastname: Cherif
authors.affiliation: Institute of Tropical Medicine (NEKKEN), Nagasaki University, Nagasaki, Japan.|Graduate School of Biomedical Sciences and Program for Nurturing Global Leaders in Tropical and Emerging Communicable Diseases, Nagasaki University, Nagasaki, Japan.
authors.forename: Saori
authors.initials: S
authors.lastname: Sakabe
authors.affiliation: Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan.
authors.forename: Ayato
authors.initials: A
authors.lastname: Takada
authors.affiliation: Institute of Tropical Medicine (NEKKEN), Nagasaki University, Nagasaki, Japan.|Graduate School of Biomedical Sciences and Program for Nurturing Global Leaders in Tropical and Emerging Communicable Diseases, Nagasaki University, Nagasaki, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Hirayama
authors.affiliation: Institute of Tropical Medicine (NEKKEN), Nagasaki University, Nagasaki, Japan.|Graduate School of Biomedical Sciences and Program for Nurturing Global Leaders in Tropical and Emerging Communicable Diseases, Nagasaki University, Nagasaki, Japan.
authors.forename: Jiro
authors.initials: J
authors.lastname: Yasuda
mesh_terms: D017931:DNA Primers / Q000235:genetics; D029043:Ebolavirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D016701:Guinea; D019142:Hemorrhagic Fever, Ebola / Q000175:diagnosis / Q000821:virology*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation / Q000379:methods*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers