MALDI-TOF mass spectrometry as a tool for differentiation of invasive and noninvasive<i>Streptococcus pyogenes</i>isolates

Moura, Hércules; Woolfitt, Adrian R.; Carvalho, Maria G. S.; Pavlopoulos, Antonis J.; Teixeira, Lúcia Martins; Satten, Glen A.; Barr, John R.

doi:10.1111/j.1574-695x.2008.00428.x

Cited by 72 publications

(47 citation statements)

References 43 publications

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“…The resulting protein spectra are postulated to be characteristic for each bacterial species. The available literature regarding the identification of and discrimination between subspecies or even single strains by MALDI-TOF MS is limited (1,24). Francisella tularensis subspecies were successfully discriminated with surface-enhanced laser desorption ionization-time of flight MS (SELDI-TOF MS) (19,26), a modified MALDI-TOF MS method which allows selective absorption of proteins on a chromatographic array surface prior to MS (31,32).…”

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confidence: 99%

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Identification of Francisella tularensis by Whole-Cell Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry: Fast, Reliable, Robust, and Cost-Effective Differentiation on Species and Subspecies Levels

et al. 2010

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Francisella tularensis, the causative agent of tularemia, is a potential agent of bioterrorism. The phenotypic discrimination of closely related, but differently virulent, Francisella tularensis subspecies with phenotyping methods is difficult and time-consuming, often producing ambiguous results. As a fast and simple alternative, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was applied to 50 different strains of the genus Francisella to assess its ability to identify and discriminate between strains according to their designated species and subspecies. Reference spectra from five representative strains of Francisella philomiragia, Francisella tularensis subsp. tularensis, Francisella tularensis subsp. holarctica, Francisella tularensis subsp. mediasiatica, and Francisella tularensis subsp. novicida were established and evaluated for their capability to correctly identify Francisella species and subspecies by matching a collection of spectra from 45 blind-coded Francisella strains against a database containing the five reference spectra and 3,287 spectra from other microorganisms. As a reference method for identification of strains from the genus Francisella, 23S rRNA gene sequencing was used. All strains were correctly identified, with both methods showing perfect agreement at the species level as well as at the subspecies level. The identification of Francisella strains by MALDI-TOF MS and subsequent database matching was reproducible using biological replicates, different culture media, different cultivation times, different serial in vitro passages of the same strain, different preparation protocols, and different mass spectrometers.

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confidence: 99%

“…Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been successfully applied to identify bacterial species (2,10,13,15,20,22,24,25) by measuring the mass of peptides and small proteins from whole cells. The resulting protein spectra are postulated to be characteristic for each bacterial species.…”

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confidence: 99%

Identification of Francisella tularensis by Whole-Cell Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry: Fast, Reliable, Robust, and Cost-Effective Differentiation on Species and Subspecies Levels

et al. 2010

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“…By further improvement of the technique, a rapid, accurate, easy-to-use, and inexpensive method has become available for identification of microorganisms (4,14,27). MALDI-TOF MS can be used for accurate and rapid identification of various microorganisms, such as Gram-positive bacteria (2,3,9,10,22,26), Enterobacteriaceae (5), nonfermenting bacteria (6,(19)(20)(21), mycobacteria (12,16,24), anaerobes (10,23), and yeasts (18,25). Most studies have reported on MALDI-TOF MS identification of a single strain or family of microorganisms in a research setting.…”

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confidence: 99%

High-Throughput Identification of Bacteria and Yeast by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry in Conventional Medical Microbiology Laboratories

2010

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Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is suitablefor high-throughput and rapid diagnostics at low costs and can be considered an alternative for conventional biochemical and molecular identification systems in a conventional microbiological laboratory. First, we evaluated MALDI-TOF MS using 327 clinical isolates previously cultured from patient materials and identified by conventional techniques (Vitek-II, API, and biochemical tests). Discrepancies were analyzed by molecular analysis of the 16S genes. Of 327 isolates, 95.1% were identified correctly to genus level, and 85.6% were identified to species level by MALDI-TOF MS. Second, we performed a prospective validation study, including 980 clinical isolates of bacteria and yeasts. Overall performance of MALDI-TOF MS was significantly better than conventional biochemical systems for correct species identification (92.2% and 83.1%, respectively) and produced fewer incorrect genus identifications (0.1% and 1.6%, respectively). Correct species identification by MALDI-TOF MS was observed in 97.7% of Enterobacteriaceae, 92% of nonfermentative Gram-negative bacteria, 94.3% of staphylococci, 84.8% of streptococci, 84% of a miscellaneous group (mainly Haemophilus, Actinobacillus, Cardiobacterium, Eikenella, and Kingella [HACEK]), and 85.2% of yeasts. MALDI-TOF MS had significantly better performance than conventional methods for species identification of staphylococci and genus identification of bacteria belonging to HACEK group. Misidentifications by MALDI-TOF MS were clearly associated with an absence of sufficient spectra from suitable reference strains in the MALDI-TOF MS database. We conclude that MALDI-TOF MS can be implemented easily for routine identification of bacteria (except for pneumococci and viridans streptococci) and yeasts in a medical microbiological laboratory.

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“…A study using an AB 4700 Proteomics Analyzer (Applied Biosystems, Foster City, USA) combined with the Tag-ident proteomics tool and the ExPASy sequence retrieval system (http://us.expasy.org) has shown that S. pyogenes isolates from necrotizing fasciitis clustered together and could be separated from noninvasive isolates, despite being the same emm types (5). However, the number of strains in the study was small (n ϭ 9) and, even using a larger collection, different emm types were not all separated by MS.…”

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Microbial Typing by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: Do We Need Guidance for Data Interpretation?

et al. 2015

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eThe integration of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) in clinical microbiology has revolutionized species identification of bacteria, yeasts, and molds. However, beyond straightforward identification, the method has also been suggested to have the potential for subspecies-level or even type-level epidemiological analyses. This minireview explores MALDI-TOF MS-based typing, which has already been performed on many clinically relevant species. We discuss the limits of the method's resolution and we suggest interpretative criteria allowing valid comparison of strain-specific data. We conclude that guidelines for MALDI-TOF MS-based typing can be developed along the same lines as those used for the interpretation of data from pulsed-field gel electrophoresis (PFGE). Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has recently been integrated into the routine diagnostic workflow of many industrial, pharmaceutical, and medical microbiology laboratories. It has already been thoroughly evaluated for the identification of clinically relevant bacterial species, including anaerobes, Gram-positive rods and cocci, Enterobacteriaceae, and miscellaneous Gram-negative (including nonfermentative) rods, with adequate to excellent results (1, 2).Species identification, however, is only a first step in the diagnostic workflow. The ability to quickly and reliably distinguish or "type" related bacterial isolates is essential for bacterial transmission studies and larger-scale epidemiological surveillance projects. "Conventional" phenotyping methods, such as multiple-susceptibility testing, phage typing, serotyping, biochemical typing, and several others, have been important contributors to our understanding of the epidemiology of community-and health careassociated infections. However, these methods all have practical limitations which render them largely unsuitable for comprehensive bacterial population analyses as well as for scientifically less ambitious but critical infection surveillance. Furthermore, most phenotypic methods have been developed for individual bacterial taxa and are not transferable to other taxa without considerable adaptation.Hence, over the past 2 decades, phenotyping has been largely replaced by "molecular" genotyping. Clonal reproduction by binary fission imprints the evolutionary history of the organism in genotypic markers amenable to analysis by nucleic acid-mediated methods. However, in practice, the ease with which recombination, transfection, and transformation can take place in bacteria necessitates that data from multiple genetic markers are analyzed in defining a "precise" genotype and even then there is no guarantee that an adequate natural taxonomy will be derived. Polyphasic taxonomy currently uses combinations of different phenotypic and/or genotypic data sets to define genera, species, and entities at or below the subspecies level. Still, all these approaches remain time-consuming and relatively...

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MALDI-TOF mass spectrometry as a tool for differentiation of invasive and noninvasiveStreptococcus pyogenesisolates

Cited by 72 publications

References 43 publications

Identification of Francisella tularensis by Whole-Cell Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry: Fast, Reliable, Robust, and Cost-Effective Differentiation on Species and Subspecies Levels

Identification of Francisella tularensis by Whole-Cell Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry: Fast, Reliable, Robust, and Cost-Effective Differentiation on Species and Subspecies Levels

High-Throughput Identification of Bacteria and Yeast by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry in Conventional Medical Microbiology Laboratories

Microbial Typing by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: Do We Need Guidance for Data Interpretation?

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