Mass spectrometry: Pneumococcal meningitis verified and Brucella species identified in less than half an hour

Hartmeyer, Gitte Nyvang; Jensen, Anne K.; Böcher, S.; Bartels, Mette Damkjær; Pedersen, Michael; Clausen, Marianne Engell; Abdul-Redha, Rawaa Jalil; Dargis, Rimtas; Schouenborg, P; Højlyng, Niels; Kemp, Michael; Christensen, Jens Jørgen

doi:10.3109/00365541003754493

Cited by 42 publications

(21 citation statements)

References 2 publications

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“…One such report describes the diagnosis of pneumococcal meningitis in a 46-yearold man (304). Following sample acquisition, the CSF was processed in a manner similar to that discussed above for urine samples: low-speed centrifugation to remove leukocytes followed by high-speed centrifugation to pellet bacteria.…”

Section: Cerebrospinal Fluidmentioning

confidence: 99%

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: Cerebrospinal Fluidmentioning

confidence: 99%

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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“…Although the Biotyper reference library does not contain select agents, Bruker's securityrelevant (SR) library does. The SR library can be obtained by users and searched simultaneously with the Biotyper reference library (6).…”

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confidence: 99%

Importance of Using Bruker's Security-Relevant Library for Biotyper Identification of Burkholderia pseudomallei, Brucella Species, and Francisella tularensis

Cunningham¹,

Patel

2013

J Clin Microbiol

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Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is an ideal platform for rapid and accurate identification of bacteria in clinical microbiology (1). Expeditious and inerrant identification of potentially hazardous agents protects laboratory staff from inadvertent exposure and enables prompt recognition of potential select agents. With the exception of experienced laboratorians who frequently encounter such agents (e.g., veterinary, public health, and reference laboratory staff), most laboratorians rely on excluding more commonly encountered bacteria before considering less common and potentially hazardous isolates. The ensuing testing may take several days, placing personnel at risk of laboratory-acquired infection.Although the standard Biotyper reference library (Bruker, Billerica, MA) does not include select agents, several groups have demonstrated the high accuracy of MALDI-TOF MS in identification of multiple species of potentially hazardous bacteria when using appropriately constructed spectral libraries. Using a supplemented Burkholderia library, Inglis et al. identified Burkholderia pseudomallei isolated from blood cultures, reducing time to identification by 24 h (2). Using a supplemented library covering Brucella species, Ferreira et al. used MALDI-TOF MS to rapidly identify clinical isolates of Brucella species (3), and using a custom-made Brucella library, Lista et al. were able to identify Brucella species to the species level using MALDI-TOF MS (4). Finally, using a supplemented library, Seibold et al. reported use of MALDI-TOF MS for differentiation of species and even subspecies of Francisella (5). Although library supplementation enhances the performance of commercial MALDI-TOF MS libraries, given challenges obtaining and working with select agents in the United States, this is not an easy endeavor. Although the Biotyper reference library does not contain select agents, Bruker's securityrelevant (SR) library does. The SR library can be obtained by users and searched simultaneously with the Biotyper reference library (6).As a large reference laboratory, we regularly encounter bacteria classified as select agents. We have historically used phenotypic tests and 16S rRNA gene sequencing to identify Brucella species, Francisella tularensis, and B. pseudomallei. Recent implementation of the Biotyper system provided an opportunity to evaluate identification of select agents with the MALDI Biotyper reference library (version 3.3.1.2; 4,612 entries) alone and with concomitant analysis against the SR library (version 1.0; 107 entries).We studied 20 isolates, including Brucella melitensis (n ϭ 6), Brucella suis (n ϭ 3), Francisella tularensis (n ϭ 9), and Burkholderia pseudomallei (n ϭ 2). Isolates were extracted in a biological safety cabinet prior to MALDI-TOF MS. Extraction was performed as previously described (7), except that the 70% ethanol exposure time was extended to 10 min to sterilize the samples (8).(This strategy will not sterilize spore-forming bacte...

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“…Demarco et al recently described a diafiltration method using desalting, fractionation, and concentration for preparing urine samples before MALDI-TOF MS analysis (58 ). Other sample types, such as cerebrospinal fluid (59,60 ), may also be tested directly.…”

Section: Direct Testing Of Clinical Samplesmentioning

confidence: 99%

MALDI-TOF MS for the Diagnosis of Infectious Diseases

2015

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BACKGROUND:First introduced into clinical microbiology laboratories in Europe, MALDI-TOF MS is being rapidly embraced by laboratories around the globe. Although it has multiple applications, its widespread adoption in clinical microbiology relates to its use as an inexpensive, easy, fast, and accurate method for identification of grown bacteria and fungi based on automated analysis of the mass distribution of bacterial proteins.CONTENT: This review provides a historical perspective on this new technology. Modern applications in the clinical microbiology laboratory are reviewed with a focus on the most recent publications in the field. Identification of aerobic and anaerobic bacteria, mycobacteria, and fungi are discussed, as are applications for testing urine and positive blood culture bottles. The strengths and limitations of MALDI-TOF MS applications in clinical microbiology are also addressed.

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Mass spectrometry: Pneumococcal meningitis verified and Brucella species identified in less than half an hour

Cited by 42 publications

References 2 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

Importance of Using Bruker's Security-Relevant Library for Biotyper Identification of Burkholderia pseudomallei, Brucella Species, and Francisella tularensis

MALDI-TOF MS for the Diagnosis of Infectious Diseases

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