Investigators enrolled 430 patients clinically suspected of meningitis who were PCR- and culture-negative for bacterial meningitis. The study ran across five tertiary hospitals in Ethiopia, selected to provide geographic representation: University of Gondar, Tikur Anbessa, Yekatit 12, ALERT, and Hawassa. CSF specimens were collected aseptically, aliquoted and stored at −80 °C, then processed using a commercial viral nucleic acid extraction kit. For RNA targets (including enteroviruses and mumps), a two-step RT‑PCR protocol was applied; DNA targets were amplified by conventional PCR according to manufacturer cycling conditions. Visualisation of amplicons used agarose gel electrophoresis. The study team used certified positive and negative controls to monitor assay performance.
Viral nucleic acid was detected in 227 of 430 participants (52.8%), investigators reported. Enteroviruses accounted for the large majority of detections: 206/430 (47.9%). Herpes simplex virus type 1 was detected in 12 patients (2.8%), HSV‑2 in 6 patients (1.4%), and Epstein‑Barr virus in 3 patients (0.7%). The laboratory screened for varicella-zoster virus and mumps virus but detected no cases for those pathogens. The authors noted that this prevalence represents the highest reported viral meningitis detection rate in Ethiopia to date and attributed part of that yield to the multicenter design and comprehensive molecular screening.
Overall, the cohort presented most commonly with fever (63%), vomiting (60.2%), lowered consciousness (43.7%), and neck stiffness (37.4%). Those proportions were higher among the PCR-confirmed viral meningitis subgroup: fever occurred in 70.9% of PCR-positive patients, vomiting in 61.7%, neck stiffness in 55.5%, and lowered consciousness in 50.7%. Among patients with enterovirus detection, fever was present in 72.8%, vomiting in 64.6%, and neck stiffness in 54.9%. For HSV‑1 positive patients, seizure activity was prominent (reported in 91.7% of HSV‑1 cases), and lowered consciousness was recorded in two-thirds of those patients. The study team observed that pediatric presentations were frequent—90.5% of participants were under 18 years—which may influence the atypicality and detection patterns of clinical signs.
Outcomes were assessed at discharge using the Glasgow Outcome Scale (GOS), dichotomized into favorable (GOS = 5) and unfavorable (GOS = 1–4). The overall rate of unfavorable outcome was 6.7% (29/430). There was one reported death, occurring in an enterovirus-positive patient. All HSV‑2 cases in this cohort had favorable outcomes. The study team ran bivariate and multivariable logistic regression analyses to identify predictors of unfavorable outcome. In the adjusted model, three factors remained independently associated with an unfavorable GOS: male sex (adjusted odds ratio reported by authors 2.684, 95% CI 1.177–6.119), longer duration from symptom onset to hospital visit (AOR 1.433, 95% CI 1.043–1.967), and a history of antibiotic intake prior to hospital presentation (AOR 2.784, 95% CI 1.270–6.104). Being PCR-positive for a viral pathogen did not show a statistically significant independent association with unfavorable outcome in this cohort.
The authors outlined quality-control measures for the molecular assays, including use of external positive controls for enterovirus, EBV, and VZV and RNase/DNase‑free water as a negative control. The cohort was predominantly pediatric, which the investigators reported as a characteristic of their sample and a potential influence on symptom patterns and outcome distribution. The study did not report detailed subtyping or molecular characterization of the enteroviruses detected; the authors explicitly called for further identification and characterization of these enteroviruses. The source did not provide additional information on long-term outcomes beyond discharge or on how antibiotic exposure prior to presentation may have altered initial clinical management. Investigators also acknowledged challenges inherent to resource-limited settings that can hinder routine viral diagnostics and emphasized the need for expanded molecular surveillance to inform clinical practice and public health response.