Microwave (MW) sensing offers noninvasive, real-time detection of the electromagnetic properties of biological materials via the highly concentrated electromagnetic fields, for which advantages include wide bandwidth, small size, and cost-effective fabrication. In this paper, we present the application of MW broadband dielectric spectroscopy (BDS) coupled to a fabricated biological thin film for evaluating ultraviolet-C (UV-C) exposure effects. The BDS thin film technique could be deployed as a biological indicator for assessing whole-room UV-C surface disinfection. The disinfection process is monitored by BDS as changes in the electrical properties of surface-confined biological thin films photodegraded with UV-C radiation. Fetal bovine serum (FBS, a surrogate for protein) and bacteriophage lambda double-stranded deoxyribonucleic acid (dsDNA) were continuously monitored with BDS during UV-C radiation exposure. The electrical resistance of FBS films yielded promising yet imprecise readings, whereas the resistance of dsDNA films discernibly decreased with UV-C exposure. The observations are consistent with the expected photo-oxidation and photodecomposition of protein and DNA. While further research is needed to characterize these measurements, this study presents the first application of BDS to evaluate the electrical properties of solid-state biological thin films. This technique shows promise toward the development of a test method and a standard biological test to determine the efficacy of UV-C disinfection. Such a test with biological indicators could easily be applied to hospital rooms between patient occupancy for a multipoint evaluation to determine if a room meets a disinfection threshold set for new patients.