Improvement of a RT-PCR assay for Yellow Fever virus genome detection
Rocha, Tatiana Carneiro da
Silva, Mario Antonio Navarro
Bona, Ana Caroline Dalla
Nunes, Márcio Roberto Teixeira
Svoboda, Walfrido Huhl
Gomes, Eliane Carneiro
The aim of the present study was to describe an improved protocol of reverse transcription polymerase chain reaction (RT-PCR) for Yellow Fever virus genome detection. A strain of ribonucleic acid of Yellow Fever virus was submitted to the improved protocol of RT-PCR and the amplicons were visualized under ultraviolet transilluminator, purifed and sequenced. The nucleotide sequence obtained was compared with sequences available in GenBank using the tblastx tool. The amplicons produced by the strain of ribonucleic acid of Yellow Fever virus exhibited fragments of 400 and 800 base pairs and the consensus sequence exhibited a similarity of 100% with Yellow Fever virus sequences recorded in GenBank. The improved protocol described in this study allowed Yellow Fever virus genome detection and enabled the elimination of the nested-PCR step, which has been frequently associated with contamination. In addition, it reduced the time of reaction, the cost of reagents and the possibility of sample contamination. New methods of investigating these infections must be elaborated and a continuous vigilance of these viruses in their diﬀerent vectors and hosts is required to avoid negative impacts on human health, tourism and trade.
xmlui.dri2xhtml.METS-1.0.item-citationROCHA, Tatiana Carneiro da et al. Improvement of a RT-PCR assay for Yellow Fever virus genome detection. Revista de Ciências Farmacêuticas Básica e Aplicada, v. 38, n. 1, p. 1-4, 2017.
xmlui.dri2xhtml.METS-1.0.item-decsPrimaryFebre Amarela / genética
Reação em Cadeia da Polimerase / métodos
Reação em Cadeia da Polimerase em Tempo Real / métodos
Flavivirus / patogenicidade
Genoma Viral / genética