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Background: Direct or indirect hand contact plays a significant role in health-care-associated infections. Family members of pediatric patients may have various hand hygiene practices. We aimed to evaluate the perceptions of hand hygiene and handshaking practices among family caregivers of hospitalized children in a tertiary care hospital. Methods: A cross-sectional study was conducted using a pretested two-part questionnaire for a randomly selected caregiver of hospitalized children at King Khalid University Hospital, Riyadh, Saudi Arabia, between January 21 and March 8, 2018. The survey data were analyzed using SPSS version 21. Results: One hundred and eighty caregivers voluntarily participated in the study. The mean age was 35.1 years, and female parents comprised 85.6% of the sample. The majority of participants (82.8%) did not receive any previous formal education on hand hygiene. Most of the participants (relative importance index of 84.6%) had correct answers on a modified version of the World Health Organization “Perception Survey for Health-Care Workers.” However, handshaking avoidance was low in general, with caregivers reporting their handshaking practices did not change even when dealing with people who have flu-like symptoms. There was no significant correlation between the participants' characteristics and handshaking avoidance practice, except for those who were working in the medical field, who showed a significantly higher handshaking avoidance. Conclusion: Formal education for hospitalized children's caregivers on hand hygiene and handshaking practice is lacking. Information on appropriate hand hygiene and potential risks of infection spread is required, especially for those parents without a previous medical background.