Real-time RT-PCR for Mayaro virus detection in plasma and urine

• Type of Material: Article
• Authors: Jesse Waggoner, Emory University; Alejandra Rojas, Universidad Nacional de Asuncion; Alisha Mohamed-Hadley, Stanford University; Yvalena Arevalo de Guillen, Universidad Nacional de Asuncion; Benjamin A. Pinsky, Stanford University
• Language: English
• Date: 2018-01-01
• Publisher: Elsevier: 12 months
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2017 Elsevier B.V.
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1016/j.jcv.2017.11.006
• Title of Journal or Parent Work: Journal of Clinical Virology
• ISSN: 1386-6532
• Volume: 98
• Start Page: 1
• End Page: 4
• Grant/Funding Information: 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 (ARS).; Research was supported by National Institutes of Health (NIH) grant K08AI110528 (JJW, salary support) and a Robert E. Shope International Fellowship in Infectious Diseases (JJW) distributed by the American Society for Tropical Medicine and Hygiene.
• Keywords: OUTBREAK; DISEASE; DENGUE; DIAGNOSIS; Virology; Science & Technology; FRENCH-GUIANA; TRANSMISSION; CHIKUNGUNYA; Mayaro virus; Quantitation; Life Sciences & Biomedicine; Plasma; Real-time RT-PCR; Urine; INFECTION
• Research Categories: Health Sciences, Public Health; Biology, Virology; Health Sciences, Pathology

Abstract

Background Mayaro virus (MAYV) causes an acute febrile illness which can be difficult to differentiate from dengue or chikungunya. MAYV RNA can be detected in plasma during the first 3–5 days of illness, but only a single rRT-PCR has been fully evaluated in the literature. Objectives To develop an rRT-PCR for MAYV and evaluate assay performance using human plasma and urine samples spiked with different MAYV strains. Study design A MAYV rRT-PCR targeting a region of the 5′UTR and nsp1 gene was designed from the alignment of all complete-genome MAYV sequences to be compatible with existing laboratory protocols. The assay was evaluated using human samples spiked with six MAYV strains, including strains from each of the three genotypes. Results The linear range of the MAYV rRT-PCR extended from 1.0 to 8.0 log10copies/μL, and the lower limit of 95% detection was 8.2 copies/μL. No detection was observed when the MAYV rRT-PCR was tested with genomic RNA from related arboviruses. The assay demonstrated linear amplification of all 6 MAYV strains when spiked into human plasma samples as well as 2 strains spiked into urine. Conclusions We report the design and evaluation of an rRT-PCR for MAYV. Given the concern for MAYV emergence in the Americas and the few molecular tests that have been evaluated in the literature, this assay should provide a useful diagnostic for patients with an acute febrile illness.