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J Virol Methods ; 193(1): 23-7, 2013 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-23692685

RESUMEN

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 °C using a real-time turbidimeter that allowed detection within 1h. 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 3h to 1h 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.


Asunto(s)
Técnicas de Amplificación de Ácido Nucleico/métodos , Fiebre Amarilla/diagnóstico , Virus de la Fiebre Amarilla/aislamiento & purificación , África , Humanos , Transcripción Reversa , Sensibilidad y Especificidad , América del Sur , Temperatura , Factores de Tiempo , Fiebre Amarilla/virología
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