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

Abstract

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.

Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry in veterinary medicine: Recent advances (2019–present)

“…Developed MALDI method for rapid screening of avian mycoplasma [114] Various fungi Validation of a free online spectral database for veterinary fungi, 90% matching success [115] MALDI-TOF=Matrix-assisted laser desorption ionization-time of flight tarda isolates were obtained from diseased fish. Then, peptides and proteins were extracted from the bacteria and used to create library reference spectra for identification.…”

“…The work demonstrates conclusively that this organism can be found in pets and thus poses a potential for transmission to humans. Baudler et al [114] reported that diagnosis of avian Mycoplasma by conventional methods can be a time-consuming task fraught with difficulty. Hence, these authors developed a MALDI-TOF Mycoplasma spectral database of 36 main spectrum profiles from 23 avian Mycoplasma spp.…”