Rotavirus is the most common cause of severe diarrhea. Worldwide prevalence of the virus is high and Nigeria is yet to adopt the World Health Organisation (WHO) recommendation for global inclusion of rotavirus vaccine into the national program on immunization due to the high cost of the vaccine. The study assessed Nigerian mothers' awareness of rotavirus infection, the importance of the rotavirus vaccine, its acceptance, and their willingness-to-pay (WTP) value for the vaccine. A quantitative study was carried out at eligible immunization units in Anambra State, using a self-administered validated questionnaire. Yamane's sample size formula was used to get the sample size and data collected from eligible mothers were collated using Microsoft Excel, 2010. Various outcomes were determined with a Chi-square test using SPSS (Ver. 20.0 for Windows, Inc., Chicago, IL, USA). Of the 376 mothers assessed, the maximum amount most of the women were willing to pay was less than 1000 naira per dose of the vaccine. WTP amount and knowledge of RV infection were statistically associated with the level of education and average total income per month of the mothers (p< 0.05). There was no statistically significant difference in outcomes observed with the different study settings. There was poor awareness of rotavirus infection among mothers as well as poor awareness of the importance of the rotavirus vaccine, but most mothers were willing to pay a maximum of 1, 000.00 Naira [$3.0] per dose for the vaccine.