Skip header navigation

University of Stirling

×

Article

Development of a single-tube one-step RT-LAMP assay to detect the Chikungunya virus genome

Alternative title Detection of Chikunguny a virus by RT-LAMP

Citation
Lopez Jimena B, Wehner S, Harold G, Bakheit M, Frischmann S, Bekaert M, Faye O, Sall AA & Weidmann M (2018) Development of a single-tube one-step RT-LAMP assay to detect the Chikungunya virus genome [Detection of Chikunguny a virus by RT-LAMP]. PLoS Neglected Tropical Diseases, 12 (5), Art. No.: e0006448. https://doi.org/10.1371/journal.pntd.0006448; https://doi.org/10.1371/journal.pntd.0006448

Abstract
Background A single-tube one-step real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid detection of chikungunya virus (CHIKV) targeting the conserved 6K-E1 target region was developed. The assay was validated with sera collected from a CHIKV outbreak in Senegal in 2015. Methodology/Principal findings A novel design approach by combining Principal Component Analysis and phylogenetic analysis of 110 available CHIKV sequences and the LAMP oligonucleotide design software LAVA was used. The assay was evaluated with an External Quality Assessment panel from the European Network for Diagnostics of "Imported" Viral Diseases and was shown to be sensitive and specific and did not cross-detect other arboviruses. The limit of detection as determined by probit analysis, was 163 molecules, and 100% reproducibility in the assays was obtained for 103 molecules (7/8 repetitions were positive for 102 molecules). The assay was validated using 35 RNA samples extracted from sera, and results were compared with those obtained by quantitative RT-PCR carried out at the Institut Pasteur Dakar, demonstrating that the RT-LAMP is 100% sensitive and 80% specific, with a positive predictive value of 97% and negative predictive value of 100%. Conclusions/Significance The RT-LAMP appeared to show superior performance with material stored for months compared to qRT-PCR and can be therefore recommended for use in infrastructures with poor settings.

Journal
PLoS Neglected Tropical Diseases: Volume 12, Issue 5

StatusPublished
Author(s)Lopez Jimena, Benjamin; Wehner, Stefanie; Harold, Graham; Bakheit, Mohammed; Frischmann, Sieghard; Bekaert, Michaël; Faye, Oumar; Sall, Amadou A; Weidmann, Manfred
FundersEuropean Commission
Publication date29/05/2018
Publication date online29/05/2018
Date accepted by journal12/04/2018
URLhttp://hdl.handle.net/1893/27312
PublisherPublic Library of Science
Publisher URLhttps://doi.org/10.1371/journal.pntd.0006448
Scroll back to the top