Generic RT-nested-PCR for detection of flaviviruses using degenerated primers and internal control followed by sequencing for specific identification

Sánchez-Seco, María Paz; Rosario, Delfina; Domingo, Cristina; Hernández, Lourdes; Valdés, Katia; Guzmán, María G.; Tenorio, Antonio

doi:10.1016/j.jviromet.2005.01.025

Cited by 122 publications

(105 citation statements)

References 27 publications

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“…A generic test (the Generic reverse transcription (RT)-nestedpolymerase chain reaction (PCR) test for fl aviviruses), followed by sequencing for specific viruses was proposed (25) , and specifi c diagnosis of ZIKV in fi eld-collected mosquitoes can be accomplished by quantitative real-time PCR (26) .…”

Section: Zika Virusmentioning

confidence: 99%

Zika virus in Brazil and the danger of infestation by Aedes (Stegomyia) mosquitoes

Marcondes

Ximenes

2015

Rev. Soc. Bras. Med. Trop.

215

157

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Zika virus, already widely distributed in Africa and Asia, was recently reported in two Northeastern Brazilian: State of Bahia and State of Rio Grande do Norte, and one Southeastern: State of São Paulo. This fi nding adds a potentially noxious virus to a list of several other viruses that are widely transmitted by Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus in Brazil. The pathology and epidemiology, including the distribution and vectors associated with Zika virus, are reviewed. This review is focused on viruses transmitted by Aedes (Stegomyia) mosquitoes, including dengue, Chikungunya, Zika, Mayaro, and yellow fever virus, to emphasize the risks of occurrence for these arboviruses in Brazil and neighboring countries. Other species of Aedes (Stegomyia) are discussed, emphasizing their involvement in arbovirus transmission and the possibility of adaptation to environments modifi ed by human activities and introduction in Brazil.

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Section: Zika Virusmentioning

confidence: 99%

Zika virus in Brazil and the danger of infestation by Aedes (Stegomyia) mosquitoes

Marcondes

Ximenes

2015

Rev. Soc. Bras. Med. Trop.

215

157

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“…1 In this region, recent studies indicate that the virus is circulating in birds, horses, and humans. [10][11][12] For the first time in Spain, disease by WNV have been described in 2010 in horses (ProMED-mail, archive no. : 20101119.4203) and humans (ProMED-mail, archive no.…”

Section: Introductionmentioning

confidence: 99%

West Nile and Usutu Viruses in Mosquitoes in Spain, 2008–2009

Vázquez¹,

Ruíz²,

Herrero³

et al. 2011

The American Society of Tropical Medicine and Hygiene

109

106

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“…Los flavivirus son una preocupación real para la salud pública (9). En este estudio se implementaron técnicas de biología molecular que permitieron el estudio de infecciones por flavivirus en individuos con sospecha clínica de dengue y resultado negativo para antígeno NS1 de DENV, provenientes del área metropolitana de Asunción, en el periodo 2011 a 2013.…”

Section: Discussionunclassified

“…La búsqueda del genoma de flavivirus se realizó a través de la reacción de RTnested-PCR descripta por Sánchez Seco et al 2005, con algunas modificaciones (SS1) (9). La retrotranscripción (RT) para la obtención del ácido desoxirribonucleico complementario (cADN) a partir del ARN previamente extraído se realizó utilizando una transcriptasa reversa Moloney Murine Leukemia Virus (M-MLV) (Promega, EEUU) y random primers (Byodinamics, AR).…”

Section: Búsqueda Del Genoma De Flavivirus Mediante Rt-nested Pcr Genunclassified

Search of flavivirus in individuals with clinical suspicion of dengue and negative for the NS1 antigen in Paraguay

Segovia

Franco

et al. 2017

Mem. Inst. Investig. Cienc. Salud

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R E S U M E NLos flavivirus son responsables de una considerable morbi-mortalidad a nivel mundial. Entre ellos, el virus del dengue (DENV) es causante de graves problemas de salud pública en Paraguay. El objetivo del estudio fue detectar infecciones por flavivirus a través de una reacción de RT-nested PCR genérica para flavivirus en 195 muestras de individuos con sospecha de dengue, negativos por el test inmunocromatográfico (antígeno NS1 -DENV), provenientes del área metropolitana de Asunción entre 2011 y 2013. Las muestras positivas para flavivirus fueron sometidas a dos reacciones de RT-nested PCRs específicas para DENV. El límite de detección (LD) para flavivirus fue de 0,2 UFP/reacción. En total 43/195 muestras fueron positivas para flavivirus. De estas, 38/43 (88,4%) correspondieron a DENV (6 DENV-1, 30 DENV-2 y 2 DENV-3). Además, 5/43 casos (11,6%) positivos para flavivirus fueron negativos para DENV por ambas reacciones específicas, pudiendo deberse a infecciones por otros flavivirus. Los resultados sugieren que la utilización de una reacción genérica seguida de otras reacciones específicas para DENV en casos febriles negativos para NS1 por el método inmunocromatográfico permitiría detectar más casos de infecciones por DENV y además, podría contribuir a la identificación de casos debido a infecciones por otros flavivirus. Flaviviruses are responsible for considerable worldwide morbidity and mortality. Among them, the dengue virus (DENV) causes serious public health problems in Paraguay. The objective of the study was to detect flavivirus infections using a generic RT-nested -PCR in 195 samples of individuals with suspected dengue and negative for

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Generic RT-nested-PCR for detection of flaviviruses using degenerated primers and internal control followed by sequencing for specific identification

Cited by 122 publications

References 27 publications

Zika virus in Brazil and the danger of infestation by Aedes (Stegomyia) mosquitoes

Zika virus in Brazil and the danger of infestation by Aedes (Stegomyia) mosquitoes

West Nile and Usutu Viruses in Mosquitoes in Spain, 2008–2009

Search of flavivirus in individuals with clinical suspicion of dengue and negative for the NS1 antigen in Paraguay

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