Real-time RT-PCR for Venezuelan equine encephalitis complex, Madariaga, and Eastern equine encephalitis viruses: application in human and mosquito public health surveillance in Panama.
Jean-Paul Carrera, Dimelza Araúz, Alejandra Rojas, Fátima Cardozo, Victoria Stittleburg, Ingra Morales Claro, Josefrancisco Galue, Carlos Lezcano-Coba, Filipe Romero Rebello Moreira, Luis Felipe -Rivera, Maria Chen-Germán, Brechla Moreno, Zeuz Capitan-Barrios, Sandra López-Vergès, Juan Miguel Pascale, Ester C Sabino, Anayansi Valderrama, Kathryn A Hanley, Christl A Donnelly, Nikos Vasilakis, Nuno R Faria, Jesse J Waggoner
Author Information
Jean-Paul Carrera: Department of Biology, University of Oxford , Oxford, United Kingdom. ORCID
Dimelza Araúz: Department of Research in Virology and Biotechnology, Gorgas Memorial Institute of Health Studies , Panama City, Panama.
Alejandra Rojas: Departamento de Producción, Instituto de Investigaciones en Ciencias de la Salud, Universidad Nacional de Asunción , San Lorenzo, Paraguay.
Fátima Cardozo: Departamento de Producción, Instituto de Investigaciones en Ciencias de la Salud, Universidad Nacional de Asunción , San Lorenzo, Paraguay.
Victoria Stittleburg: Division of Infectious Diseases, Department of Medicine, Emory University , Atlanta, Georgia, USA.
Ingra Morales Claro: Instituto de Medicina Tropical, Faculdade de Medicina da Universidade de São Paulo , São Paulo, Brazil.
Josefrancisco Galue: Department of Research in Virology and Biotechnology, Gorgas Memorial Institute of Health Studies , Panama City, Panama.
Carlos Lezcano-Coba: Department of Research in Virology and Biotechnology, Gorgas Memorial Institute of Health Studies , Panama City, Panama.
Filipe Romero Rebello Moreira: MRC Centre for Global Infectious Disease Analysis (MRC-GIDA), Department of Infectious Disease Epidemiology, Imperial College London , London, United Kingdom.
Luis Felipe -Rivera: Department of Research in Virology and Biotechnology, Gorgas Memorial Institute of Health Studies , Panama City, Panama.
Maria Chen-Germán: Department of Research in Virology and Biotechnology, Gorgas Memorial Institute of Health Studies , Panama City, Panama.
Brechla Moreno: Department of Research in Virology and Biotechnology, Gorgas Memorial Institute of Health Studies , Panama City, Panama.
Zeuz Capitan-Barrios: Department of Research in Virology and Biotechnology, Gorgas Memorial Institute of Health Studies , Panama City, Panama.
Sandra López-Vergès: Department of Research in Virology and Biotechnology, Gorgas Memorial Institute of Health Studies , Panama City, Panama.
Juan Miguel Pascale: Clinical of Tropical Diseases and Research Unit, Gorgas Memorial Institute of Health Studies , Panama City, Panama.
Ester C Sabino: Instituto de Medicina Tropical, Faculdade de Medicina da Universidade de São Paulo , São Paulo, Brazil.
Anayansi Valderrama: Viral Emerging Disease Dynamics Group, Gorgas Memorial Institute of Health Studies , Panama City, Panama.
Kathryn A Hanley: Department of Biology, New Mexico State University , Las Cruces, New Mexico, USA.
Christl A Donnelly: Pandemic Sciences Institute, Nuffield Department of Medicine, University of Oxford , Oxford, United Kingdom.
Nikos Vasilakis: Department of Pathology, The University of Texas Medical Branch , Galveston, Texas, USA.
Nuno R Faria: Department of Biology, University of Oxford , Oxford, United Kingdom. ORCID
Jesse J Waggoner: Division of Infectious Diseases, Department of Medicine, Emory University , Atlanta, Georgia, USA.
Eastern equine encephalitis virus (EEEV), Madariaga virus (MADV), and Venezuelan equine encephalitis virus complex (VEEV) are New World alphaviruses transmitted by mosquitoes. They cause febrile and sometimes severe neurological diseases in human and equine hosts. Detecting them during the acute phase is hindered by non-specific symptoms and limited diagnostic tools. We designed and clinically assessed real-time reverse transcription polymerase chain reaction assays (rRT-PCRs) for VEEV complex, MADV, and EEEV using whole-genome sequences. Validation involved 15 retrospective serum samples from 2015 to 2017 outbreaks, 150 mosquito pools from 2015, and 118 prospective samples from 2021 to 2022 surveillance in Panama. The rRT-PCRs detected VEEV complex RNA in 10 samples (66.7%) from outbreaks, with one having both VEEV complex and MADV RNAs. VEEV complex RNA was found in five suspected dengue cases from disease surveillance. The rRT-PCR assays identified VEEV complex RNA in three () pools, leading to VEEV isolates in two. Phylogenetic analysis revealed the VEEV ID subtype in positive samples. Notably, 11.9% of dengue-like disease patients showed VEEV infections. Together, our rRT-PCR validation in human and mosquito samples suggests that this method can be incorporated into mosquito and human encephalitic alphavirus surveillance programs in endemic regions.
