Bacterial endophthalmitis is the most severe form of vision-threatening ocular infection. Mycoplasma pneumoniae is a primary respiratory pathogen that has been associated with several extrapulmonary manifestation, including ocular infection. In this work, we presented the microbiology diagnosis of an endophalmitis by M. pneumoniae using a novel A.R.I. WELL D-ONE® (CPM Scientifica, Italy) commercial micromethod in a Cuban pediatric patient. After positive M. pneumoniae confirmation, treatment with levofloxacin was initiated and successfully eliminated the ocular infection.