2026-06-14T03:30:13Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/108772024-09-27T21:49:38Zcom_20.500.12105_2052com_20.500.12105_2051col_20.500.12105_19609
Repisalud
author
Davo, Laura
author
Labiod, Nuria
author
Roiz, David
author
Gómez-Díaz, Elena
author
Figuerola, Jordi
author
Vazquez, Ana
author
Hernandez, Lourdes
author
Herrero-Romero, Laura
author
Sánchez-Seco, María Paz
funder
Instituto de Salud Carlos III
funder
Unión Europea. Comisión Europea
funder
Ministerio de Economía y Competitividad (España)
2020-08-10T08:33:03Z
2020-08-10T08:33:03Z
2020
Parasit Vectors . 2020 May 29;13(1):270.
10.1186/s13071-020-04110-5
1756-3305
Parasites & Vectors
32471505
http://hdl.handle.net/20.500.12105/10877
Background: Granada virus belongs to the genus Phlebovirus within the Naples serocomplex and was detected for the first time in sand flies from Spain in 2003. Seroprevalence studies have revealed that Granada virus may infect humans with most cases being asymptomatic. Moreover, recent studies in vector samples revealed that the related Massilia and Arrabida phleboviruses could be also circulating in Spain. The objective of this study was to develop and assess a new sensitive real-time RT-PCR assay for Granada virus diagnosis able to detect the related phleboviruses Massilia and Arrabida.
Methods: Two specific primers and one unique probe to detect Granada, Massilia and Arrabida viruses, without differentiating between them, were designed targeting the conserved L-segment of their genome. Sensitivity was assessed using 10-fold serial dilutions of quantified in vitro DNA samples. Specificity was evaluated by testing different genomic RNA extracted from other representative phleboviruses. The new assay was used for virus detection in sand flies collected in 2012 from the Balearic Archipelago, a touristic hotspot in the Mediterranean.
Results: The real-time RT-PCR assay exhibited a sensitivity per reaction of 19 copies for Granada and Arrabida, and 16 copies for Massilia. No other related phleboviruses were detected. From the 37 pools of sand fly samples studied from four different Balearic Islands, we detected one positive in the island of Cabrera.
Conclusions: To our knowledge, the method described here is the first real-time RT-PCR designed to detect Granada virus and the related Massilia and Arrabida phleboviruses. The study demonstrated that this is a rapid, robust and reliable assay for the accurate diagnosis of human infections as well as for virus surveillance in vectors.
eng
Real-time RT-PCR assay to detect Granada virus and the related Massilia and Arrabida phleboviruses
journal article
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URL
https://repisalud.isciii.es/bitstreams/eae5a9ef-c4d2-477c-918a-6e4580a035ee/download
File
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application/pdf
RealTimeRTPCR_2020.pdf
URL
https://repisalud.isciii.es/bitstreams/596a03f1-020c-4fbb-8ffc-f33d19c28507/download
File
MD5
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text/plain
RealTimeRTPCR_2020.pdf.txt