Please use this identifier to cite or link to this item: http://hdl.handle.net/1893/29235
Appears in Collections:Aquaculture Journal Articles
Peer Review Status: Refereed
Title: Development, validation and clinical evaluation of a broad-range pan-filovirus RT-qPCR
Author(s): Jääskeläinen, Anne J
Sironen, Tarja
Diagne, Cheikh Tidiane
Diagne, Moussa Moïse
Faye, Martin
Faye, Oumar
Faye, Ousmane
Hewson, Roger
Mölsä, Markos
Weidmann, Manfred W
Watson, Robert
Sall, Amadou Alpha
Vapalahti, Olli
Contact Email: m.w.weidmann@stir.ac.uk
Keywords: Ebola
Marburg
Sudan
Bundibugyo
Pan-Filo
Issue Date: May-2019
Citation: Jääskeläinen AJ, Sironen T, Diagne CT, Diagne MM, Faye M, Faye O, Faye O, Hewson R, Mölsä M, Weidmann MW, Watson R, Sall AA & Vapalahti O (2019) Development, validation and clinical evaluation of a broad-range pan-filovirus RT-qPCR. Journal of Clinical Virology, 114, pp. 26-31. https://doi.org/10.1016/j.jcv.2019.03.010
Abstract: Background During the five decades since their discovery, filoviruses of four species have caused human hemorrhagic fever outbreaks: Marburg (MARV) marburgvirus, and Zaire (EBOV), Sudan (SUDV) and Bundybugyo (BDBV) ebolaviruses. The largest, devastating EBOV epidemic in West Africa in 2014-16, has been followed by outbreaks of MARV in Uganda, 2017, and EBOV in Democratic Republic of Congo, 2018, emphasizing the need to develop preparedness to diagnose all filoviruses. Objectives The aim of this study was to optimize a new filovirus RT-qPCR to detect all filoviruses, define its limits of detection (LOD) and perform a field evaluation with outbreak samples. Study design A pan-filovirus RT-qPCR targeting the L gene was developed and evaluated within the EbolaMoDRAD (Ebola virus: modern approaches for developing bedside rapid diagnostics) project. Specificity and sensitivity were determined and the effect of inactivation and PCR reagents (liquid and lyophilized format) were tested. Results The LODs for the lyophilized pan-filovirus L-RT-qPCR assay were 9.4 copies per PCR reaction for EBOV, 9.9 for MARV, 1151 for SUDV, 65 for BDBV and 289 for Taï Forest virus. The test was set at the Pasteur Institute, Dakar, Senegal, and 83 Ebola patient samples, with viral load ranging from 5 to 5 million copies of EBOV per reaction, were screened. The results for the patient samples were in 100% concordance with the reference EBOV-specific assay. Discussion Overall, the assay showed good sensitivity and specificity, covered all filoviruses known to be human pathogens, performed well both in lyophilized and liquid-phase formats and with EBOV outbreak clinical samples.
DOI Link: 10.1016/j.jcv.2019.03.010
Rights: This item has been embargoed for a period. During the embargo please use the Request a Copy feature at the foot of the Repository record to request a copy directly from the author. You can only request a copy if you wish to use this work for your own research or private study. Accepted refereed manuscript of: Jääskeläinen AJ, Sironen T, Diagne CT, Diagne MM, Faye M, Faye O, Faye O, Hewson R, Mölsä M, Weidmann MW, Watson R, Sall AA & Vapalahti O (2019) Development, validation and clinical evaluation of a broad-range pan-filovirus RT-qPCR. Journal of Clinical Virology, 114, pp. 26-31. DOI: https://doi.org/10.1016/j.jcv.2019.03.010 © 2019, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/

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