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PLoS Negl Trop Dis. 2016 Feb 22;10(2):e0004472. doi: 10.1371/journal.pntd.0004472. eCollection 2016 Feb.

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.

Author information

1
Institute of Tropical Medicine (NEKKEN), Nagasaki University, Nagasaki, Japan.
2
Laboratoire des Fièvres Hémorragiques en Guinée, Conakry, Guinea.
3
Hôpital National Donka, Service des Maladies Infectieuses et Tropicales, Conakry, Guinea.
4
Université Gamal Abdel Nasser, Conakry, Guinea.
5
Graduate School of Biomedical Sciences and Program for Nurturing Global Leaders in Tropical and Emerging Communicable Diseases, Nagasaki University, Nagasaki, Japan.
6
Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan.

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.

PMID:
26900929
PMCID:
PMC4764121
DOI:
10.1371/journal.pntd.0004472
[Indexed for MEDLINE]
Free PMC Article

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