Rapid identification of Burkholderia mallei and Burkholderia pseudomalleiby intact cell Matrix-assisted Laser Desorption/Ionisation mass spectrometric typing

Karger, Axel; Stock, Rüdiger; Ziller, Mario; Elschner, Mandy C.; Bettin, Barbara; Melzer, Falk; Maier, Τ.; Kostrzewa, Markus; Scholz, Holger C.; Neubauer, Heinrich; Tomaso, Herbert

doi:10.1186/1471-2180-12-229

Cited by 59 publications

(47 citation statements)

References 40 publications

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“…Additionally, the authors reported that spectra generated from B. mallei exhibited higher levels of homogeneity than did those from B. pseudomallei, with the type strain of B. pseudomallei being separated from more recent isolates of B. pseudomallei. This may be due to genetic modification of the type strain due to significant passage or inappropriate medium combinations (205). This finding represents a potential challenge with regard to standardization of specimen age and levels of manipulation of microbes submitted for MALDI-TOF MS identification.…”

Section: Finally Mencacci Et Al Recognized the Potential Of Maldimentioning

confidence: 98%

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: a Fundamental Shift in the Routine Practice of Clinical Microbiology

et al. 2013

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SUMMARY Within the past decade, clinical microbiology laboratories experienced revolutionary changes in the way in which microorganisms are identified, moving away from slow, traditional microbial identification algorithms toward rapid molecular methods and mass spectrometry (MS). Historically, MS was clinically utilized as a high-complexity method adapted for protein-centered analysis of samples in chemistry and hematology laboratories. Today, matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) MS is adapted for use in microbiology laboratories, where it serves as a paradigm-shifting, rapid, and robust method for accurate microbial identification. Multiple instrument platforms, marketed by well-established manufacturers, are beginning to displace automated phenotypic identification instruments and in some cases genetic sequence-based identification practices. This review summarizes the current position of MALDI-TOF MS in clinical research and in diagnostic clinical microbiology laboratories and serves as a primer to examine the “nuts and bolts” of MALDI-TOF MS, highlighting research associated with sample preparation, spectral analysis, and accuracy. Currently available MALDI-TOF MS hardware and software platforms that support the use of MALDI-TOF with direct and precultured specimens and integration of the technology into the laboratory workflow are also discussed. Finally, this review closes with a prospective view of the future of MALDI-TOF MS in the clinical microbiology laboratory to accelerate diagnosis and microbial identification to improve patient care.

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Section: Finally Mencacci Et Al Recognized the Potential Of Maldimentioning

confidence: 98%

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: a Fundamental Shift in the Routine Practice of Clinical Microbiology

et al. 2013

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“…Overall, novel skills for particular and rapid identification of microorganisms are a very important step toward a appropriate treatment of infectious diseases in veterinary and medical diagnostics and are of major concern today (Hays and Leeuwen, 2012). Identification of different pathogens with mass spectrometry has been known as a technique for acurate and quick recognation of bacteria (Giebel et al, 2010;Karger et al, 2012). Consequently, the most commonly applied technique for mass spectral identification of microorganisms is Matrix Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF-MS) (Biswas and Rolain, 2013).…”

Section: Introductionmentioning

confidence: 99%

Phenotypical and Mass Spectral Assessment Methods for Identification of Some Contagious Mastitis Pathogens

Behiry¹,

Zahran²,

Marzouk³

et al. 2014

American Journal of Microbiology

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Mastitis is one of the most economic disease affecting dairy cows worldwide. Identification of mastitis pathogens still depends principally on culture and phenotypical method, which is a difficult and timeconsuming. Newly, microbiologists have focused their attention on the use of Mass Spectrometry (MS) for microbial identification, especially Matrix Assisted Laser Desorption Ionization Time-Of-Flight (MALDI-TOF). Therefore, this study was designated to evaluate the ability of MALDI-TOF to identify some contagious mastitis pathogens comparing with phenotypical methods such as API panels and VITEK 2 system. A total of one hundred twenty of Staphylococcus aureus (S. aureus), Coagulase Negative Staphylococci (CNS) and Streptococcus agalactiae (Strept. agalactiae) strains isolated from milk of cows affected by clinical and subclinical mastitis were used in the study. According to the results, ~95% of S. aureus, 100% of CNS and Strept. agalactiae were correctly identified by MALDI TOF MS. All S. aureus isolates were then confirmed by a nuc-based PCR technique. While ~92% of S. aureus, 87% of Strept. agalactiae and 76% of CNS were identified by VITEK 2 system. Moreover, ~89% of S. aureus, 80% of Strept. agalactiae and 72% of CNS were identified by API system. In brief, the results demonstrated that MALDI-TOF is a fast and truthful technique which has the capability to replace conventional identification of several bacterial strains usually isolated in clinical laboratories of microbiology. Therefore, it is recomended that MALDI-TOF MS technology can be regularly used in veterinary laboratories for identification of different species of bacteria, particularly when failure of phenotypic methods forces clinical microbiologists.

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“…B. pseudomallei matched Biotyper scores at the species-level threshold (Ͼ2.0) based on the Bruker database, which was only slightly higher than that for the related SSBA bacterial species, Burkholderia mallei ( Table 1). The local database produced higher match scores for B. pseudomallei, and we plan to expand the local MSP entries, as previous studies have recommended the use of an appropriate supplemental reference library for the genus Burkholderia (17).…”

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confidence: 99%

Verification of a Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Method for Diagnostic Identification of High-Consequence Bacterial Pathogens

2016

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bWe examined the utility of a single matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry method for the identification of security-sensitive biological agents (risk group 3 bacterial pathogens). The goal was 2-fold: to verify a method for inclusion into our scope of accreditation, and to assess the biological safety of extractions. We developed our sample flow to include a tube-based chemical extraction, followed by filtration, before processing on MALDI-TOF MS instruments in a containment level 2 laboratory. Matrix-assisted laser desorption-ionization time of flight mass spectrometry (MALDI-TOF MS) is a rapid, robust, and cost-effective technology, which is emerging as a routine bacterial identification method in clinical diagnostic laboratories (1-3). Despite its utility, there are concerns with the safety of sample processing and the quality of spectral databases for the identification of security-sensitive biological agents (SSBAs). As our laboratory predominately works with SSBA bacterial pathogens (the risk group 3 [RG-3] pathogens: Francisella tularensis, Yersinia pestis, Bacillus anthracis, Brucella species, and Burkholderia pseudomallei), our goal was to verify the usefulness and safety of a MALDI-TOF MS method for inclusion into our scope of accreditation (ISO 17025). We chose to pursue a full-tube-based protein extraction method involving cellular disruption and filtration, rather than the direct-colony plate (smear) method (4, 5), in an effort to mitigate the risk of viable organisms.Over a period of 1 year, 146 bacterial samples (including 57 SSBA samples; Table 1) were processed as previously described (6). The bacteria were previously identified by standard genotypic and phenotypic methods. In brief, all cultures were chemically extracted in a microcentrifuge tube using 70% ethanol, 70% formic acid, and acetonitrile, and all SSBA bacteria were extracted in a biosafety level 3 (BSL-3) laboratory. SSBA extracts from sporeforming bacteria were filtered through a 0.1 M microcentrifuge filtration unit (EMD Millipore, Etobicoke, Ontario, Canada), as recommended by Dauphin and Bowen (7). The viability of the extract was performed by inoculating 10% of the extract onto both sheep's blood agar (SBA) (or cysteine heart agar [CHA] for F. tularensis) and into 2 ml of tryptic soy broth (TSB), and incubating at 35°C and 5% CO 2 for 48 h. After 48 h, the TSB was further subcultured to SBA or CHA and monitored for an additional 72 h. Any bacterial growth was tested with agent-specific assays to rule out the SSBA, indicating a contaminant. After a lack of growth was confirmed, extracts were removed from the BSL-3 laboratory, and 1.5 l of the extract was applied (at a minimum of four replicate spots) to an MSP-96 MALDI target plate (Bruker Daltonics, East Milton, Ontario, Canada), with an overlay of 1.5 l of ␣-cyano-4-hydroxycinnamic acid (CHCA) matrix solution (50% acetonitrile and 2.5% trifluoroacetic acid). Mass spectra were acquired with the FlexControl software (version 3.0...

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Rapid identification of Burkholderia mallei and Burkholderia pseudomalleiby intact cell Matrix-assisted Laser Desorption/Ionisation mass spectrometric typing

Cited by 59 publications

References 40 publications

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: a Fundamental Shift in the Routine Practice of Clinical Microbiology

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: a Fundamental Shift in the Routine Practice of Clinical Microbiology

Phenotypical and Mass Spectral Assessment Methods for Identification of Some Contagious Mastitis Pathogens

Verification of a Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Method for Diagnostic Identification of High-Consequence Bacterial Pathogens

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