A real-time reverse transcription loop-mediated isothermal amplification assay for the rapid detection of yellow fever virus
Autor(es): Kwallah Allan ole, Inoue Shingo, Muigai Anne W T, Kubo Toru, Sang Rosemary, Morita Kouichi, Mwau Matilu
Resumo: Yellow fever, a mosquito-borne disease, is an important viral hemorrhagic fever in Africa - 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 - 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 - other related viruses where only YFV RNA was detected: West Nile virus, dengue viruses, Japanese encephalitis virus, Rift Valley fever virus, - chikungunya virus. In addition, equal sensitivity was also observed when the RT-LAMP - 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 - increased sensitivity tenfold compared to RT-PCR. Therefore, this test offers a simple, rapid - reliable diagnostic tool for yellow fever when there are outbreaks of acute hemorrhagic fever in Kenya - other African countries.
Palavras-Chave: NS1; RT-LAMP; YFV
Imprenta: Journal of Virological Methods, v. 193, n. 1, p. 23-27, 2013
Identificador do objeto digital: 10.1016/j.jviromet.2013.05.004
Descritores: Chikungunya virus - Flaviviridae ; Chikungunya virus - Pathogenesis ; Chikungunya virus - RNA ; Chikungunya virus - Infectious diseases ; Chikungunya virus - Viral infections ; Chikungunya virus - PCR detection ; Chikungunya virus - Real Time PCR ; Chikungunya virus - RT-PCR ; Chikungunya Virus - Virus ; Chikungunya virus - Vaccine ; Chikungunya virus - Chikungunya fever ; Chikungunya virus - Dengue ; Chikungunya virus - Public health
Data de publicação: 2013
