Mycoplasma hominis can cause life-threatening central nervous system infections in neonates following intrauterine infection or during delivery. In newborns, the dia gnosis and treatment of M. hominis meningitis are challenging, because cultures are often negative and the bacterium is not susceptible to empirical antibiotics. Herein, we describe a case of neonatal M. hominis meningitis diagnosed using real-time polymerase chain reaction (RT-PCR) and treated with ciprofloxacin. The patient was a 3-day-old female hospitalized for a fever and lethargy. Her blood laboratory findings were non-specific; cerebrospinal fluid (CSF) examination showed a white blood cell count of 580/µL and indicated meningitis. Her symptoms could not be controlled with empirical antibiotics. Urine culture on a special medium revealed ciprofloxacinsusceptible M. hominis. Furthermore, the RT-PCR performed with the CSF sample revealed M. hominis. Therefore, the patient was administered cipro­floxacin; after 2 days, the fever subsided. The patient was discharged on day 30 without complications.