Implementation of a Multiplex rRT-PCR for Zika, Chikungunya, and Dengue Viruses: Improving Arboviral Detection in an Endemic Region

• Type of Material: Article
• Authors: César Cantero Cantero, Universidad Nacional de Asunción; Fatima Cardozo, Universidad Nacional de Asunción; Jesse Waggoner, Emory University; Benjamin A. Pinsky, Stanford University; Anibal Espinola, Universidad Nacional de Asunción; Belen Infanzon, Universidad Nacional de Asunción; Maria Eugenia Acosta, Universidad Nacional de Asunción; Laura Aria, Universidad Nacional de Asunción; Yvalena Arevalo de Guillen, Universidad Nacional de Asunción; Teresa Cuevas, Universidad Nacional de Asunción; Vicenta Rojas, Universidad Nacional de Asunción; Clotilde Segovia, Universidad Nacional de Asunción; Ana Centurion, Universidad Nacional de Asunción; Alejandra Rojas, Universidad Nacional de Asunción
• Language: English
• Date: 2020-03-01
• Publisher: AMER SOC TROP MED & HYGIENE
• Publication Version: Final Published Version
• Copyright Statement: © The American Society of Tropical Medicine and Hygiene
• Final Published Version (URL): http://dx.doi.org/10.4269/ajtmh.19-0707
• Title of Journal or Parent Work: AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE
• Volume: 102
• Issue: 3
• Start Page: 625
• End Page: 628
• Grant/Funding Information: The research was 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.). The research was also supported by a grant from the Dirección General de Investigación Científica y Tecnológica, Rectorado, Universidad Nacional Asunción (DGICT-UNA).
• Keywords: Public, Environmental & Occupational Health; Science & Technology; Tropical Medicine; VIREMIA; Life Sciences & Biomedicine
• Research Categories: Health Sciences, Pathology

Abstract

Arboviral diagnosis has been complicated throughout the tropical and subtropical Americas by the recent co-circulation of Zika virus (ZIKV), chikungunya virus (CHIKV), and dengue virus (DENV). The aim of this study was to implement a multiplex real-time RT-PCR (rRT-PCR) for ZIKV, CHIKV, and DENV in Paraguay to test patients who were clinically suspected of having dengue. We tested 110 sera from patients who presented to the Hospital de Clínicas in 2016 and had testing for DENV nonstructural protein 1 (NS1; 40 positive and 70 negative). Using a composite reference standard, we confirmed 51 dengue cases (46.4%): 38/40 NS1 positive and 13/70 NS1 negative. Chikungunya virus and ZIKV were detected in one sample each, both were DENV NS1 negative. The NS1 test demonstrated good agreement with rRT-PCR for DENV. However, multiplex rRT-PCR identified a subset of dengue cases and additional arboviral infections that would not be detected if NS1 assays are relied upon for diagnosis.