Neurovirus Emerging in the Americas Study (NEAS): Neurologic Manifestations of the Arbovirus Dengue, Chikungunya and Zika Infections in Colombia

NEAS is led by Dr. Carlos Pardo-Villamizar at Johns Hopkins University (JHU) and Drs. Beatriz Parra and Lyda Osorio at Universidad del Valle (UV) in Colombia. They have established a collaborative network with physicians, neurologists and experts in microbiology and epidemiology in that country. NEAS is actively recruiting patients in seven university-based hospitals in five Colombian cities. The protocol of the study has been approved in each of the participating centers.

NEAS prospective case-control study aims to define the association between the exposure to arbovirus (Zika, Dengue and Chikungunya) and the development of a neurological syndrome in patients from Colombia, a country with endemic transmission of arbovirus. The cases are defined as individuals that present with new onset of a neurological syndrome of unknown etiology, including but not limited to encephalitis, myelitis, meningitis, polyneuropathy/Guillain-Barre syndrome and cranial nerve involvement. There are two age-matched control groups: 1) household controls and 2) controls with a febrile syndrome of unknown etiology that do not present neurological involvement. The estimated sample size in the study is 50 participants in each group (cases and controls 1 and 2). The time to collect the complete sample size is between 12-24 months, and depends on the local incidence of cases.

Each center has a coordinator team that includes a neurologist, a physician - neurology or internal medicine resident -, and a research coordinator. The team is constantly screening the patients admitted to the center. The patients that fulfill the criteria for cases or controls are approached by the research coordinator who is in charge to inform the participant about the study and complete the informed consent. Then, the medical team perform the neurological evaluation, including the history and the physical exam according to the NEAS forms. Samples of blood and urine are collected in all participants. If a lumbar puncture is done as part of the standard of care, spinal fluid is also collected. Based on resources availability, the participants are followed every week for a 4-week period and in each visit a clinical assessment and sample collection are completed.

Demographic and clinical information, laboratory and imaging results are stored in a secured web-based database in the platform REDcap,based at JHU. Each center has a unique combination of a username and a password to enter the database and include the information of each participant in real time. Each center has access to the included information by the same center and no by other centers. Standardized forms (NEAS forms) were created by the study team that include relevant information necessary to achieved the aims of this study and guarantee the data consistency. Laboratory testing and imaging results are obtained from the medical records. A permanent monitoring of the data base is done by the teams at JHU and UV.

After the samples are collected they are shipped to the study core laboratory located at UV (Dr. Beatriz Parra). Each center has been trained in the correct handling, processing, and shipping of the samples. Once the samples are in the core laboratory, they are aliquoted and stored. Blood, urine and spinal fluid are tested by Real Time/Reverse Transcriptase (RT)- Polymerase Chain Reaction (PCR) for the presence of viral RNA (Ribonucleic Acid) of Zika virus and Dengue virus. Serum and spinal fluid are tested by ELISA for the detection of anti-flavivirus IgM and IgG. The results are shared with the local study team and with the medical team taking care of the participant. In addition, aliquots of samples are sent to JHU for additional immunological and virological assessments (Dr. Carlos Pardo-Vllamizar).