Rapid Diagnostic Tests as a Source of Dengue Virus RNA for Envelope Gene Amplification: A Proof of Concept

Sarah Cassidy-Seyoum 1, 2 Manivanh Vongsouvath 1 Onanong Sengvilaipaseuth 1 Malee Seephonelee 1 Tehmina Bharucha 1, 3 Xavier de Lamballerie 4 Paul Newton 1, 5, 2 Audrey Dubot-Pérès 4, 1, 5
1 LOMWRU - Lao-Oxford-Mahosot Hospital-Wellcome Trust Research Unit
2 LSHTM - London School of Hygiene and Tropical Medicine
3 UCL - University College of London [London]
4 UVE - Unité des Virus Emergents
5 Nuffield Department of Clinical Medicine [Oxford]
Abstract : Molecular epidemiological data are key for dengue outbreak characterization and preparedness. However, sparse Dengue virus (DENV) molecular information is available in Laos because of limited resources. In this proof-of-concept study, we evaluated whether DENV1 RNA extracted from rapid diagnostic tests (RDTs) could be amplified and sequenced. The protocol for envelope gene amplification from RNA purified from RDTs was first assessed using viral isolate dilutions then conducted using 14 dengue patient sera. Envelope gene amplification was successful from patient sera with high virus titer, as was sequencing but with lower efficiency. Hence, based on our results, RDTs can be a source of DENV1 RNA for subsequent envelope gene amplification and sequencing. This is a promising tool for collecting molecular epidemiology data from rural dengue-endemic areas. However, further investigations are needed to improve assay efficiency and to assess this tool's level of efficacy on a larger scale in the field.
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Sarah Cassidy-Seyoum, Manivanh Vongsouvath, Onanong Sengvilaipaseuth, Malee Seephonelee, Tehmina Bharucha, et al.. Rapid Diagnostic Tests as a Source of Dengue Virus RNA for Envelope Gene Amplification: A Proof of Concept. American Journal of Tropical Medicine and Hygiene, American Society of Tropical Medicine and Hygiene, 2019, 101 (2), pp.451-455. ⟨10.4269/ajtmh.18-0831⟩. ⟨inserm-02375424⟩

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