Internally Controlled, Multiplex Real-Time Reverse Transcription PCR for Dengue Virus and Yellow Fever Virus Detection

• Type of Material: Article
• Authors: Alehandta Rojas, Universidad Nacional de Asunción; Cheikh T. Diagne, Institut Pasteur de Dakar; Victoria D. Stittleburg, Emory University; Alisha Mohamed-Hadley, Stanford University; Yvalena Arevalo de Guillen, Universidad Nacional de Asunción; Angel Balmaseda, Ministry of Health, Nicaragua; Oumar Faye, Institut Pasteur de Dakar; Ousmane Faye, Institut Pasteur de Dakar; Amadou A. Sall, Institut Pasteur de Dakar; Eva Harris, Sustainable Sciences Institute, Nicaragua; Benjamin A. Pinsky, Stanford University; Jesse Waggoner, Emory University
• Language: English
• Date: 2018-01-01
• Publisher: American Society of Tropical Medicine and Hygiene
• Publication Version: Final Published Version
• Copyright Statement: Copyright © 2018 by The American Society of Tropical Medicine and Hygiene.
• Final Published Version (URL): http://dx.doi.org/10.4269/ajtmh.18-0024
• Title of Journal or Parent Work: American Journal of Tropical Medicine and Hygiene
• ISSN: 0002-9637
• Volume: 98
• Issue: 6
• Start Page: 1833
• End Page: 1836
• Grant/Funding Information: The research was supported by the National Institutes of Health (NIH) grant K08AI110528 (J. J. W., salary support); and a Robert E. Shope International Fellowship in Infectious Diseases (J. J. W.) distributed by the American Society of Tropical Medicine and Hygiene.; Research was also supported by a fellowship from the Consejo Nacional de Ciencia y Tecnología (CONACYT) of Paraguay, awarded as part of the Programa de Vinculación de Científicos y Tecnólogos, PVCT 16-66 (A. R.).
• Keywords: GENOME; DIAGNOSIS; Life Sciences & Biomedicine; Science & Technology; Public, Environmental & Occupational Health; ASSAY; Tropical Medicine; INFECTIOUS-DISEASES
• Research Categories: Biology, Virology; Health Sciences, Public Health; Health Sciences, Medicine and Surgery

Abstract

The differential diagnosis of dengue virus (DENV) and yellow fever virus (YFV) infections in endemic areas is complicated by nonspecific early clinical manifestations. In this study, we describe an internally controlled, multiplex real-time reverse transcription polymerase chain reaction (rRT-PCR) for the detection of DENV and YFV. The DENV–YFV assay demonstrated specific detection and had a dynamic range of 2.0–8.0 log10 copies/µL of eluate for each DENV serotype and YFV. Clinical performance was similar to a published pan-DENV assay: 48/48 acute-phase samples from dengue cases were detected in both assays. For YFV detection, mock samples were prepared with nine geographically diverse YFV isolates over a range of concentrations. The DENV–YFV assay detected 62/65 replicates, whereas 54/65 were detected using a reference YFV rRT-PCR. Given the reemergence of DENV and YFV in areas around the world, the DENV–YFV assay should be a useful tool to narrow the differential diagnosis and provide early case detection.