Hotel room cleanliness is based on observation and not on microbial assessment even though recent reports suggest that infections may be acquired while staying in hotel rooms. Exploratory research in the first part of the authors’ study was conducted to determine if contamination of hotel rooms occurs and whether visual assessments are accurate indicators of hotel room cleanliness. Data suggested the presence of microbial contamination that was not reflective of visual assessments. Unfortunately, no standards exist for interpreting microbiological data and other indicators of cleanliness in hotel rooms. The purpose of the second half of the authors’ study was to examine cleanliness standards in other industries to see if they might suggest standards in hotels. Results of the authors’ study indicate that standards from other related industries do not provide analogous criteria, but do provide suggestions for further research.