Analysis of a reverse transcription loop-mediated isothermal amplification (RT-LAMP) for yellow fever diagnostic

• Autor(a) principal: Nunes, Márcio Roberto Teixeira
• Data de Publicação: 2015
• Outros Autores: Vianez Júnior, João Lídio da Silva Gonçalves, Nunes, Keley Nascimento Barbosa, Silva, Sandro Patroca da, Lima, Clayton Pereira Silva de, Guzman, Hilda, Martins, Lívia Caricio, Carvalho, Valéria Lima, Tesh, Robert B, Vasconcelos, Pedro Fernando da Costa
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Digital do Instituto Evandro Chagas (Patuá)
• Texto Completo: https://patua.iec.gov.br/handle/iec/2760
• topic: Vírus da Febre Amarela / isolamento & purificação; Febre Amarela / diagnóstico; Febre Amarela / virologia; Transcrição Reversa; Reação em Cadeia da Polimerase Via Transcriptase Reversa / métodos; Técnicas de Amplificação de Ácido Nucleico / métodos

Yellow Fever virus (YFV) is an important human pathogen in tropical areas of Africa and South America. Although an efficient vaccine is available and has been used since the early 1940s, sylvatic YFV transmission still occurs in forested areas where anthropogenic actions are present, such as mineral extraction, rearing livestock and agriculture, and ecological tourism. In this context, two distinct techniques based on the RT-PCR derived method have been previously developed, however both methods are expensive due to the use of thermo cyclers and labeled probes. We developed isothermal genome amplification, which is a rapid, sensitive, specific and low cost molecular approach for YFV genome detection. This assay used a set of degenerate primers designed for the NS1 gene and was able to amplify, within 30 min in isothermal conditions, the YFV 17D vaccine strain derived from an African wild prototype strain (Asibi),as well as field strains from Brazil, other endemic countries from South and Central America, and the Caribbean. The generic RT-LAMP assay could be helpful for YFV surveillance in field and rapid response during outbreaks in endemic areas.