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

Autor: Nunes MR; Center for Technological Innovation, Evandro Chagas Institute, Brazilian Ministry of Health, Ananindeua, Pará, Brazil. Electronic address: marcionunes@iec.pa.gov.br., Vianez JL Jr; Center for Technological Innovation, Evandro Chagas Institute, Brazilian Ministry of Health, Ananindeua, Pará, Brazil., Nunes KN; Center for Technological Innovation, Evandro Chagas Institute, Brazilian Ministry of Health, Ananindeua, Pará, Brazil., da Silva SP; Center for Technological Innovation, Evandro Chagas Institute, Brazilian Ministry of Health, Ananindeua, Pará, Brazil., Lima CP; Center for Technological Innovation, Evandro Chagas Institute, Brazilian Ministry of Health, Ananindeua, Pará, Brazil., Guzman H; Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA., Martins LC; Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, Brazilian Ministry of Health, Ananindeua, Pará, Brazil., Carvalho VL; Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute, Brazilian Ministry of Health, Ananindeua, Pará, Brazil., Tesh RB; Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA., Vasconcelos PF; 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.
Jazyk: angličtina
Zdroj: Journal of virological methods [J Virol Methods] 2015 Dec 15; Vol. 226, pp. 40-51. Date of Electronic Publication: 2015 Oct 13.
DOI: 10.1016/j.jviromet.2015.10.003
Abstrakt: 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.
(Copyright © 2015 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: