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J Virol Methods. 2015 Dec 15;226:40-51. doi: 10.1016/j.jviromet.2015.10.003. Epub 2015 Oct 13.

Analysis of a Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) for yellow fever diagnostic.

Author information

1
Center for Technological Innovation, Evandro Chagas Institute, Brazilian Ministry of Health, Ananindeua, Pará, Brazil. Electronic address: marcionunes@iec.pa.gov.br.
2
Center for Technological Innovation, Evandro Chagas Institute, Brazilian Ministry of Health, Ananindeua, Pará, Brazil.
3
Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA.
4
Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, Brazilian Ministry of Health, Ananindeua, Pará, Brazil.
5
Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, Brazilian Ministry of Health, Ananindeua, Pará, Brazil; Department of Pathology, University of Para State, Belem, Brazil.

Abstract

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.

KEYWORDS:

Diagnostic; Genome detection; RT-LAMP; Yellow fever virus

PMID:
26459206
DOI:
10.1016/j.jviromet.2015.10.003
[Indexed for MEDLINE]
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