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Detection of  Entamoeba histolytica  and its differentiation from  Entamoeba dispar  is an important goal of the clinical parasitology laboratory. The aim of this study was the identification and differentiation of  E .  histolytica  and  E .  dispar  by MALDI-TOF MS, in order to evaluate the application of this technique in routine diagnostic practice. MALDI-TOF MS was applied to 3 amebic reference strains and to 14 strains isolated from feces that had been differentiated by molecular methods in our laboratory. Protein extracts from cultures of these strains (axenic cultures for the 3 reference strains and monoxenic cultures for the 14 field isolates) were analyzed by MALDI-TOF MS and the spectra obtained were analyzed by statistical software. Five peaks discriminating between  E .  histolytica  and  E .  dispar  reference strains were found by protein profile analysis: 2 peaks (8,246 and 8,303 Da) specific for  E .  histolytica  and 3 (4,714; 5,541; 8,207 Da) for  E .  dispar . All clinical isolates except one showed the discriminating peaks expected for the appropriate species. For 2 fecal samples from which 2 strains (1  E .  histolytica  and 1  E .  dispar ) out of the 14 included in this study were isolated, the same discriminating peaks found in the corresponding isolated amebic strains were detected after only 12h ( E .  histolytica ) and 24h ( E .  dispar ) of incubation of the fecal samples in Robinson’s medium without serum. Our study shows that MALDI-TOF MS can be used to discriminate between  E .  histolytica  and  E .  dispar  using  in vitro  xenic cultures and it also could have potential for the detection of these species in clinical samples.

MALDI-TOF Mass Spectrometry for the Detection and Differentiation of Entamoeba histolytica and Entamoeba dispar