Extracts of Plumeria obtusa are widely used in ethnomedicine and have been investigated for a variety of biological activities; however, the antimicrobial activity of P. obtusa flowers is poorly characterized. In this study, the antimicrobial activities of different solvents (petroleum ether, ethyl acetate, chloroform, isobutanol and ethanol) extracts from flowers of P. obtusa were investigated by a disc diffusion method against Gram-positive bacteria, Gram-negative bacteria and a fungus. All extracts exhibited growth inhibition of all microorganisms at variable degrees as measured by relative zones of inhibition, however, the petroleum ether extract was ineffective against Klebsiella pneumonia and ethyl acetate and isobutanol extracts were ineffective against Pseudomonas aeruginosa. The most susceptible Gram-positive bacterium was Bacillus subtilis while the most resistant Gram-positive bacterium was Staphylococcus aureus. Erwinia carotovora was the most susceptible Gram-negative bacterium while P. aeruginosa was highly resistant among the Gram-negative bacteria. In this study, for the first time, we investigated the antimicrobial activity of several different solvent extracts from flowers of P. obtusa against a broad spectrum of human-pathogenic microorganisms. These compounds warrant further investigation by isolation and structural elucidation with the aim to find novel and affordable bioactive compounds for the treatment of infectious diseases.