This article is under the CC BY license (https://creativecommons.org/licenses/by/4.0/). As we have seen from the current COVID-19 pandemic, misconceptions concerning viruses can lead to disease spread and panic. Therefore, it is imperative to determine misconceptions held concerning epidemics and pandemics. One virus that warrants study of misconceptions, albeit given limited attention in the West, is the Ebola virus. An assessment of college students’ knowledge and misconceptions about the Ebola virus was created and validated using data from 203 non-science majors at a Midwestern United States university. The data were analyzed using both classical and Rasch measurement methods to make a case for the validity of the assessment and to explore students’ misconceptions. The assessment was shown to be a valid and useful measure for students’ knowledge and misconceptions concerning Ebola. Integrating a confidence scale into students’ responses made the scale more reliable and assisted in identifying students’ tenacious misconceptions. Students displayed multiple misconceptions about viruses, including confusion between the characteristics of viruses and prokaryotes. Students also displayed misconceptions about Ebola itself, including the overestimation of the number of Ebola strains and the number of patients who experience massive blood loss, misunderstandings about the incubation period, and overestimation of the mortality in comparison to other diseases like Influenza and Anthrax. This assessment can be used as a starting point in future studies to determine what misconceptions people have about Ebola and which types of educational and behavioral interventions need to be undertaken.