Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 193, Issue 1, Pages 23-27Publisher
ELSEVIER
DOI: 10.1016/j.jviromet.2013.05.004
Keywords
YFV; RT-LAMP; NS1
Funding
- Consortium for National Health Research [RLG-09/19-001]
- Ministry of Education, Culture, Sports, Science and Technology of Japan
- 21st Century Centers of Excellence program on Global Strategies (GCOE) for the Control of Tropical Emerging and Infectious Diseases at Nagasaki University
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Yellow fever, a mosquito-borne disease, is an important viral hemorrhagic fever in Africa and South America where it is endemic. Detection of yellow fever virus (YFV) in Africa remains a challenge due to a lack of highly specific tests. The aim of this study was to develop and optimize a rapid detection reverse transcription loop-mediated isothermal amplification (RT-LAMP) for YFV. The RT-LAMP was done isothermally at 62 degrees C using a real-time turbidimeter that allowed detection within 1 h. Specificity of the RT-LAMP was determined using RNA from flaviviruses and other related viruses where only YFV RNA was detected: West Nile virus, dengue viruses, Japanese encephalitis virus, Rift Valley fever virus, and chikungunya virus. In addition, equal sensitivity was also observed when the RT-LAMP and the real-time RT-PCR were compared using YFV-spiked human serum samples with a detection limit of 0.29 PFU/ml. Two Kenyan YFV wild strains showed an equal detection limit as the vaccine strain 17D in this study. The RT-LAMP reduced the time of reaction from 3 h to 1 h and increased sensitivity tenfold compared to RT-PCR. Therefore, this test offers a simple, rapid and reliable diagnostic tool for yellow fever when there are outbreaks of acute hemorrhagic fever in Kenya and other African countries. (c) 2013 Elsevier B.V. All rights reserved.
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