Identification and differentiation of avian Mycoplasma species using MALDI-TOF MS.

The identification of avian Mycoplasma spp. by conventional immunologic, phenotypic, and molecular methods can be demanding and time-consuming. We evaluated MALDI-TOF MS for its suitability to identify avian mycoplasmas at the species level. We generated a mycoplasma spectral database of 36 main spectrum profiles (MSPs) representing 23 avian Mycoplasma spp. using 23 type and reference strains, 1 live vaccine strain, and 8 clinical isolates. We then used 112 avian Mycoplasma clinical isolates of different avian mycoplasmas, 4 Mycoplasma live vaccine strains, and 1 Mycoplasma type strain, previously cultured and identified to the species level by molecular methods, to evaluate the MSP database. Protein extraction and MALDI-TOF MS analysis were performed with a maximum of 3 repetitions per isolate. MALDI-TOF MS resulted in accurate species-level identification with a score of ≥2.0 for 112 of 117 (96%) isolates. The MALDI-TOF MS analysis of 4 of 5 isolates that did not yield a score of ≥2.0 resulted in best-match identifications that were still concordant at species level with the molecular method used for previous identification. Therefore, MALDI-TOF MS is a promising tool for reliable identification of avian Mycoplasma spp.