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This study was conducted to explore knowledge, attitudes, and beliefs about cervical cancer, cervical cancer screening, human papillomavirus (HPV), and acceptance of the HPV vaccine. A purposive sample of 40 women was interviewed during August 2012. Fisher’s exact test was used to evaluate differences among rural and urban women, and open-ended questions were coded independently by two individuals (Cohen’s kappa coefficient of 0.816). Among the 22 rural and 18 urban women, there was limited knowledge about cervical cancer, screening, HPV, and the HPV vaccine. Cervical cancer was described in language related to gender, science, severity, or associated with having children, a uterus, or menstruation. All rural and most urban participants were interested in the HPV vaccine for themselves and their daughters. Limited awareness and knowledge about cervical cancer and HPV was common among Guatemalan women, highlighting the need for additional information prior to developing cancer prevention educational materials and programs.