Please use this identifier to cite or link to this item: http://hdl.handle.net/1893/31450
Appears in Collections:Aquaculture Journal Articles
Peer Review Status: Refereed
Title: Comparison of Zaire ebolavirus realtime RT-PCRs targeting the nucleoprotein gene
Author(s): Jääskeläinen, Anne J
Sironen, Tarja
Kaloinen, Minttu
Kakkola, Laura
Julkunen, Ilkka
Hewson, Roger
Weidmann, Manfred
Mirazimi, Ali
Watson, Robert
Vapalahti, Olli
Contact Email: m.w.weidmann@stir.ac.uk
Keywords: Ebola
Zaire
PCR
nucleoprotein
Issue Date: Oct-2020
Citation: Jääskeläinen AJ, Sironen T, Kaloinen M, Kakkola L, Julkunen I, Hewson R, Weidmann M, Mirazimi A, Watson R & Vapalahti O (2020) Comparison of Zaire ebolavirus realtime RT-PCRs targeting the nucleoprotein gene. Journal of Virological Methods, 284, Art. No.: 113941. https://doi.org/10.1016/j.jviromet.2020.113941
Abstract: In last five years, the Africa has faced two outbreaks of Zaire ebolavirus. These outbreaks have been the largest so far, and latest outbreak is still ongoing and affecting the Democratic Republic of the Congo. We tested in parallel three different Zaire ebolavirus (EBOV) realtime RT-PCRs targeting the nucleoprotein gene (EBOV NP-RT-qPCRs) described by Trombley et al. (2010), Huang et al. (2012) and Weidmann et al. (2004). These assays are used regularly in diagnostic laboratories. The limit of detection (LOD), intra-assay repeatability using different matrixes, sensitivity and specificity were determined. In addition, the primers and probes were aligned with the sequences available in ongoing and past outbreaks in order to check the mismatches. The specificity of all three EBOV NP-RT-qPCRs were excellent (100%), and LODs were under or 10 copies per PCR reaction. Intra-assay repeatability was good in all assays, however the Ct-values were bit higher using the EDTA-blood based matrix. All of the primers and probes in EBOV NP-RTqPCR assays have one or more mismatches in the probes and primers when the 2267 Zaire EBOV NP sequences, including strains Ituri from DRC outbreak (year 2018), was aligned. The EBOV strain of Bikoro (year 2018) circulating in DRC was 100% match in Trombley and Weidmann assay, but had one mismatch in Huang assay.
DOI Link: 10.1016/j.jviromet.2020.113941
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, Kaloinen M, Kakkola L, Julkunen I, Hewson R, Weidmann M, Mirazimi A, Watson R & Vapalahti O (2020) Comparison of Zaire ebolavirus realtime RT-PCRs targeting the nucleoprotein gene. Journal of Virological Methods, 284, Art. No.: 113941. https://doi.org/10.1016/j.jviromet.2020.113941 © 2020, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/
Licence URL(s): http://creativecommons.org/licenses/by-nc-nd/4.0/

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