Ribeiro, Guilherme de Sousa. Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil. Documento produzido em parceria ou por autor vinculado à Fiocruz, mas não consta à informação no documento.Siqueira, A. M. Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil. Documento produzido em parceria ou por autor vinculado à Fiocruz, mas não consta à informação no documento.Lourenço-de-Oliveira, R.; Rosa-Freitas, M. G. Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Mosquitos Transmissores de Hematozoários, Rio de Janeiro, RJ, Brasil. Documento produzido em parceria ou por autor vinculado à Fiocruz, mas não consta à informação no documento.1 Unité des Virus Émergents (UVE: Aix-Marseille Univ-IRD 190-Inserm 1207-IHU Méditerranée Infection), Marseille, France. 2 EA7310, Laboratoire de Virologie, Université de Corse-Inserm, Corte, France. 3 Centre for Infectious Disease Control, National Institute for Public Health and the Environment (RIVM), Bilthoven, the Netherlands 4 Department Viroscience, Erasmus University Medical Center, Rotterdam, The Netherlands. 5 Institute for Human Infections and Immunity and Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, USA. 6 Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Institute of Virology, 10117 Berlin, Germany. 7 German Centre for Infection Research (DZIF), Germany. 8 Blood Systems Research Institute, San Francisco, and Department of Laboratory Medicine, University of California, San Francisco, USA. 9 Department of Pediatrics, Division of Infectious Diseases, Stanford University School of Medicine, Stanford, USA. 10 Departments of Medicine, Molecular Microbiology, Pathology and Immunology, and The Andrew M. and Jane M. Bursky Center for Human Immunology and Immunotherapy Programs, Washington University School of Medicine, St. Louis, USA. 11 Department of Pathology, Institute of Human Infection and Immunity, University of Texas Medical Branch, Galveston, USA. 12 Zoonotic Diseases and Special Pathogens, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Canada. 13 Instituto Nacional de Infectologia Evandro Chagas - Oswaldo Cruz Foundation, Rio de Janeiro, Brasil. 14 Gonçalo Moniz Institute, Oswaldo Cruz Foundation, and Federal University of Bahia, Salvador, Brazil. 15 MRC-University of Glasgow Centre for Virus Research, Glasgow, UK. 16 Institut Pasteur, Biology of Infection Unit; Inserm U1117; Paris Descartes University, Departement of Infectious Diseases and Tropical Medicine, Necker-Enfants Malades University Hospital, APHP, IHU Imagine, Paris, France. 17 Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR), Singapore. 18 Établissement Français du Sang Alpes Méditerranée, Marseille, France. 19 Medical Entomology Platform, Institut Pasteur du Cambodge, Phnom Penh, Cambodia. 20 Unité d'Entomologie Médicale, Institut Pasteur de Dakar, Dakar, Senegal. 21 Department of Virology, Institut Pasteur, Arboviruses and Insect Vectors Unit, Paris, France. 22 Section Clinical Tropical Medicine, Department of Infectious Diseases, Heidelberg University Hospital, Heidelberg, Germany. 23 Instituto Oswaldo Cruz-Fiocruz, Laboratório de Mosquitos Transmissores de Hematozoários, Rio de Janeiro, Brasil. 24 KU Leuven, Department of Microbiology and Immunology, Rega Institute for Medical Research, Laboratory of Virology and Chemotherapy, Leuven, Belgium. 25 Division of Emerging and Transfusion Transmitted Diseases, Laboratory of Emerging Pathogens, Office of Blood Research and Review, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, USA. 26 Public Health and Infection Research Group, Faculty of Health Sciences, Universidad Tecnologica de Pereira, Pereira, Colombia. 27 Blood Systems Research Institute, San Francisco, USA, and Department of Pathology and Laboratory Medicine, University of California, San Francisco, San Francisco, USA. 28 Laveran Military Teaching Hospital, Marseille, France. 29 Laboratory of Vector Control Research, Environment and Health Unit, Institut Pasteur de la Guadeloupe, Guadeloupe, France.The GloPID-R (Global Research Collaboration for Infectious Disease Preparedness) Chikungunya (CHIKV), O'nyong-nyong (ONNV) and Mayaro virus (MAYV) Working Group is investigating the natural history, epidemiology and medical management of infection by these viruses, to identify knowledge gaps and to propose recommendations for direct future investigations and rectification measures. Here, we present the first report dedicated to diagnostic aspects of CHIKV, ONNV and MAYV. Regarding diagnosis of the disease at the acute phase, molecular assays previously described for the three viruses require further evaluation, standardized protocols and the availability of international standards representing the genetic diversity of the viruses. Detection of specific IgM would benefit from further investigations to clarify the extent of cross-reactivity among the three viruses, the sensitivity of the assays, and the possible interfering role of cryoglobulinaemia. Implementation of reference panels and external quality assessments for both molecular and serological assays is necessary. Regarding sero-epidemiological studies, there is no reported high-throughput assay that can distinguish among these different viruses in areas of potential co-circulation. New specific tools and/or improved standardized protocols are needed to enable large-scale epidemiological studies of public health relevance to be performed. Considering the high risk of future CHIKV, MAYV and ONNV outbreaks, the Working Group recommends that a major investigation should be initiated to fill the existing diagnostic gaps.