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

Marcio R T Nunes, João Lídio Vianez, Keley N B Nunes, Sandro Patroca da Silva, Clayton P S Lima, Hilda Guzman, Lívia C. Martins, Valéria L. Carvalho, Robert B. Tesh, Pedro F C Vasconcelos

Research output: Contribution to journalArticle

8 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)40-51
Number of pages12
JournalJournal of Virological Methods
Volume226
DOIs
StatePublished - Dec 15 2015

Fingerprint

Yellow fever virus
Yellow Fever
Reverse Transcription
South America
Vaccines
Genome
Central America
Livestock
Agriculture
Disease Outbreaks
Minerals
Brazil
Costs and Cost Analysis
Polymerase Chain Reaction
Genes

Keywords

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

ASJC Scopus subject areas

  • Virology

Cite this

Nunes, M. R. T., Vianez, J. L., Nunes, K. N. B., da Silva, S. P., Lima, C. P. S., Guzman, H., ... Vasconcelos, P. F. C. (2015). Analysis of a Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) for yellow fever diagnostic. Journal of Virological Methods, 226, 40-51. https://doi.org/10.1016/j.jviromet.2015.10.003

Analysis of a Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) for yellow fever diagnostic. / Nunes, Marcio R T; Vianez, João Lídio; Nunes, Keley N B; da Silva, Sandro Patroca; Lima, Clayton P S; Guzman, Hilda; Martins, Lívia C.; Carvalho, Valéria L.; Tesh, Robert B.; Vasconcelos, Pedro F C.

In: Journal of Virological Methods, Vol. 226, 15.12.2015, p. 40-51.

Research output: Contribution to journalArticle

Nunes, MRT, Vianez, JL, Nunes, KNB, da Silva, SP, Lima, CPS, Guzman, H, Martins, LC, Carvalho, VL, Tesh, RB & Vasconcelos, PFC 2015, 'Analysis of a Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) for yellow fever diagnostic', Journal of Virological Methods, vol. 226, pp. 40-51. https://doi.org/10.1016/j.jviromet.2015.10.003
Nunes, Marcio R T ; Vianez, João Lídio ; Nunes, Keley N B ; da Silva, Sandro Patroca ; Lima, Clayton P S ; Guzman, Hilda ; Martins, Lívia C. ; Carvalho, Valéria L. ; Tesh, Robert B. ; Vasconcelos, Pedro F C. / Analysis of a Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) for yellow fever diagnostic. In: Journal of Virological Methods. 2015 ; Vol. 226. pp. 40-51.
@article{23b404f5d7594d93acc00ced01354f59,
title = "Analysis of a Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) for yellow fever diagnostic",
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",
author = "Nunes, {Marcio R T} and Vianez, {Jo{\~a}o L{\'i}dio} and Nunes, {Keley N B} and {da Silva}, {Sandro Patroca} and Lima, {Clayton P S} and Hilda Guzman and Martins, {L{\'i}via C.} and Carvalho, {Val{\'e}ria L.} and Tesh, {Robert B.} and Vasconcelos, {Pedro F C}",
year = "2015",
month = "12",
day = "15",
doi = "10.1016/j.jviromet.2015.10.003",
language = "English (US)",
volume = "226",
pages = "40--51",
journal = "Journal of Virological Methods",
issn = "0166-0934",
publisher = "Elsevier",

}

TY - JOUR

T1 - Analysis of a Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) for yellow fever diagnostic

AU - Nunes, Marcio R T

AU - Vianez, João Lídio

AU - Nunes, Keley N B

AU - da Silva, Sandro Patroca

AU - Lima, Clayton P S

AU - Guzman, Hilda

AU - Martins, Lívia C.

AU - Carvalho, Valéria L.

AU - Tesh, Robert B.

AU - Vasconcelos, Pedro F C

PY - 2015/12/15

Y1 - 2015/12/15

N2 - 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.

AB - 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.

KW - Diagnostic

KW - Genome detection

KW - RT-LAMP

KW - Yellow fever virus

UR - http://www.scopus.com/inward/record.url?scp=84944716467&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84944716467&partnerID=8YFLogxK

U2 - 10.1016/j.jviromet.2015.10.003

DO - 10.1016/j.jviromet.2015.10.003

M3 - Article

VL - 226

SP - 40

EP - 51

JO - Journal of Virological Methods

JF - Journal of Virological Methods

SN - 0166-0934

ER -