UV light-emitting diodes (UV-LEDs) offer various wavelength options, while microorganisms have spectral sensitivity, or so-called action spectra, which can be different among species. Accordingly, matching properly the emission spectra of UV-LEDs and the spectral sensitivity of microorganisms is a reasonable strategy to enhance inactivation. In this study, UV-LEDs with nominal peak emissions at 265, 280 and 300 nm were applied to pathogens including Legionella pneumophila, Pseudomonas aeruginosa, Vibrio parahaemolyticus and feline calicivirus, in comparison with indicator species including Escherichia coli, Bacillus subtilis spores, bacteriophage Qβ and MS2. The results indicated that, for all species tested, 265 nm UV-LED was highest in the fluence-based inactivation rate constant k, followed by 280 nm and 300 nm was much lower. The k value at 280 nm was close to that at 265 nm for feline calicivirus and MS2, suggesting that 280 nm UV-LED can be as good an option as 265 nm UV-LED to inactivate these viruses. Bacteria tended to show fluence-response curves with shoulder and tailing, while viruses followed log-linear profiles at all wavelengths tested. This study indicates the fluence-response profiles and the fluence required for a target inactivation of microorganisms, which would serve as reference data for future study and applications of UV-LEDs.