Year-end Sale % OFF 
Abstract
Chikungunya was introduced into the Americas in 2015 causing a pandemic across the continent. Testing during the acute phase of infection relies on qRT-PCR, but available assays have a number of limitations. A qRT-PCR assay specific to the chikungunya E1 gene was designed using sequence data from contemporary strains. A probit analysis established the 95% limit of detection as 19.6 copies per reaction. We compared the assay with a US Centers for Disease Control (CDC) chikungunya qRT-PCR as the reference standard. The assay had a sensitivity and specificity of 98.4% and 100% in 90 samples retrospectively collected in Guatemala. In a further 74 febrile samples prospectively collected in Ecuador and Guatemala the test had a sensitivity and specificity of 100% and 98.4%, respectively. Sequencing the nsp4 gene of the discordant positive sample indicated the presence of chikungunya RNA, and mismatches to the primer binding sites of the CDC assay.
Highlights
Chikungunya virus (CHIKV), an alphavirus transmitted by Aedes mosquitoes, causes an acute febrile illness with a wide range of symptoms including fever, rash, and headache
We evaluated its limits of detection (LOD), linearity and reproducibility using RNA extracted from cultured virus, which was quantified via qRT-PCR
The LOD of our assay lies over one log10 below the range of the viral titres reported in the Caribbean (Charles et al, 2015; Gallian et al, 2014) and is comparable to reported CHIKV qRT-PCR assays reporting LODs ranging from 5.3 to 27 RNA copies per reaction (Santiago et al, 2013)
Summary
Chikungunya virus (CHIKV), an alphavirus transmitted by Aedes mosquitoes, causes an acute febrile illness with a wide range of symptoms including fever, rash, and headache. In 2013, the Asian lineage of the CHIKV strain was reported in the island of St. Martin in the Americas. Initial reports were followed by a rapid spread across the Caribbean, causing over 790,000 cases. ☆ Sources of funding: The study was funded through the MRC Confidence in Concept award number MC-PC_14111. The funders had no role in the design of the study, data collection, analysis or preparation of the manuscript. ☆☆ Conflicts of interest statement: Clément Larcher is employed by QIAGEN Manchester Ltd, a commercial company producing diagnostics. QIAGEN, Germany, provided free kits for RNA extraction for the project and loaned an RGQ 6000
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
