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

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

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.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Primers / genetics
  • Ebolavirus / classification
  • Ebolavirus / genetics*
  • Ebolavirus / isolation & purification
  • Guinea
  • Hemorrhagic Fever, Ebola / diagnosis
  • Hemorrhagic Fever, Ebola / virology*
  • Humans
  • Nucleic Acid Amplification Techniques / instrumentation
  • Nucleic Acid Amplification Techniques / methods*
  • Reverse Transcription
  • Sensitivity and Specificity

Substances

  • DNA Primers

Grant support

This work was funded by the Ministry of Health, Labor, and Welfare of Japan, and the Japan Agency for Medical Research and Development (AMED). Ministry of Health, Labor, and Welfare of Japan: http://www.mhlw.go.jp/stf/seisakunitsuite/bunya/hokabunya/kenkyujigyou/index.html Japan Agency for Medical Research and Development: http://www.amed.go.jp/en/program/. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.