Identification and Subtyping of Clinically Relevant Human and Ruminant Mycoplasmas by Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

Abstract

g Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) recently emerged as a technology for the identification of bacteria. In this study, we aimed to evaluate its applicability to human and ruminant mycoplasmal identification, which can be demanding and time-consuming when using phenotypic or molecular methods. In addition, MALDI-TOF MS was tested as a subtyping tool for certain species. A total of 29 main spectra (MSP) from 10 human and 13 ruminant mycoplasma (sub)species were included in a mycoplasma MSP database to complete the Bruker MALDI Biotyper database. After broth culture and protein extraction, MALDI-TOF MS was applied for the identification of 119 human and 143 ruminant clinical isolates that were previously identified by antigenic or molecular methods and for subcultures of 73 ruminant clinical specimens that potentially contained several mycoplasma species. MALDI-TOF MS resulted in accurate (sub)species-level identification with a score of >1.700 for 96% (251/262) of the isolates. The phylogenetically closest (sub)species were unequivocally distinguished. Although mixtures of the strains were reliably detected up to a certain cellular ratio, only the predominant species was identified from the cultures of polymicrobial clinical specimens. For typing purposes, MALDI-TOF MS proved to cluster Mycoplasma bovis and Mycoplasma agalactiae isolates by their year of isolation and genome profiles, respectively, and Mycoplasma pneumoniae isolates by their adhesin P1 type. In conclusion, MALDI-TOF MS is a rapid, reliable, and cost-effective method for the routine identification of high-density growing mycoplasmal species and shows promising prospects for its capacity for strain typing.

A case report of Mycoplasma hominis brain abscess identified by MALDI-TOF mass spectrometry

“…This has also been shown previously by Pereyre et al who successfully applied this technique for the identification of Mycoplasma spp, including M. hominis isolates. 6 In our case of M. hominis CNS infection, MALDI-TOF MS was a reliable tool and the use of 16S rDNA sequencing could have been avoided.…”

Loss of diversity within Mycoplasma bovis isolates collected in France from bovines with respiratory diseases over the last 35 years

“…Protein extracts were prepared as described previously from 1 ml of 48 h broth cultures (Pereyre et al, 2013). The final pellets were dissolved in an equal volume (20 ll) of 70% formic acid and acetonitrile.…”

“…However DNA rearrangements were shown to occur within the vsp loci in clonal populations (Buchenau et al, 2010;Lysnyansky et al, 1996) so their use for subtyping strains with an epidemiological concordance was limited. In contrast the overall protein patterns obtained by MatrixAssisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) were shown to be promising for subtyping strains of M. bovis (Pereyre et al, 2013).…”

“…Two molecular subtyping techniques, that were expected to cover the diversity of strains in case this was extensive (MLST) or limited (MLVA), were used and optimized to answer this biological question. Strains were also analyzed by MALDI-TOF MS to confirm the preliminary results obtained in an evaluation of the performance of MALDI-TOF MS in identifying mycoplasmas (Pereyre et al, 2013).…”

Novel Strategy for Typing Mycoplasma pneumoniae Isolates by Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Coupled with ClinProTools

“…Recently, a study using MALDI-TOF MS to identify and type Mycoplasma genus members found that 96% of Mycoplasma strains can be identified at the species level. For typing purposes, MALDI-TOF MS was shown to cluster M. bovis and M. agalactiae isolates by their year of isolation and genome profile, respectively, and M. pneumoniae isolates by their adhesion p1 type using the MALDI-TOF Biotyper software (28). Here, a novel strategy for typing type 1 and type 2 M. pneumoniae strains using the MALDI-TOF MS ClinProTools system was developed.…”