The aim of this work was to report the molecular diagnosis of Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum' in a domestic feline. Mycoplasma spp. are pleomorphic bacteria that parasitize the surface of red blood cells in several domestic species, mainly cats, being responsible for feline mycoplasmosis, which can cause hemolytic anemia. Cats can become infected through blood transfusions, social interactions via fights and flea bites. In this report, the patient exhibited in the clinical exam, pale and icteric mucous membranes, and enlargement of the popliteal and submandibular lymph nodes. Based on the symptoms presented, a blood sample was collected to search for hemoparasites using blood smears and also molecular examination by polymerase chain reaction (PCR). The blood smear revealed inclusions characteristic of Mycoplasma spp. Observed in red blood cells, and the diagnosis of mycoplasmosis was confirmed by PCR. This report highlights the importance of screening for hemoparasites in the feline clinical routine, with molecular tests being the most recommended due to their high sensitivity and specificity.