Use of Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Analysis for Rapid Confirmation of Burkholderia pseudomallei in Septicemic Melioidosis

Inglis, Timothy J. J.; Healy, Paul E.; Fremlin, Leith J.; Golledge, C.L.

doi:10.4269/ajtmh.2012.11-0454

Cited by 35 publications

(30 citation statements)

References 12 publications

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“…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).…”

mentioning

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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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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“…Confirmed B. pseudomallei were then transferred to Perth, Western Australia for detailed characterization. Isolates were sent as suspensions of bacteria in sterile water and duplicated in nutrient agar-embedded bacteriology [13][14][15] Confirmed B. pseudomallei were also subjected to biogeographic attribution PCR assays for Yersinia-like fimbrial (YLF) and Burkholderia thailandensis-like flagellum and chemotaxis (BTFC) gene clusters. 16 Molecular epidemiology.…”

Section: Methodsmentioning

confidence: 99%

“…MALDI-TOF mass spectra were analyzed using proprietary software (Biotyper; Bruker Daltonic GmbH, Bremen, Germany), the manufacturer's bacterial database and an extended in-house Burkholderia species reference library. 13 MLST ST profiles were analyzed and displayed using goeBURST with single-locus variants (SLVs) selected. 18 Available STs and country of origin were obtained from the MLST website and details filled in from original descriptions.…”

Section: Methodsmentioning

confidence: 99%

Sri Lankan National Melioidosis Surveillance Program Uncovers a Nationwide Distribution of Invasive Melioidosis

Corea¹,

Merritt²,

Ler³

et al. 2016

The American Society of Tropical Medicine and Hygiene

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Abstract. The epidemiologic status of melioidosis in Sri Lanka was unclear from the few previous case reports. We established laboratory support for a case definition and started a nationwide case-finding study. Suspected Burkholderia pseudomallei isolates were collated, identified by polymerase chain reaction assay, referred for Matrix Assisted Laser Desorption Ionization-Time of Flight analysis and multilocus sequence typing (MLST), and named according to the international MLST database. Between 2006 and early 2014, there were 32 patients with culture-confirmed melioidosis with an increasing annual total and a falling fatality rate. Patients were predominantly from rural communities, diabetic, and male. The major clinical presentations were sepsis, pneumonia, soft tissue and joint infections, and other focal infection. Burkholderia pseudomallei isolates came from all parts of Sri Lanka except the Sabaragamuwa Province, the south central hill country, and parts of northern Sri Lanka. Bacterial isolates belonged to 18 multilocus sequence types, one of which (ST 1137) was associated with septicemia and a single-organ focus (Fisher's exact, P = 0.004). Melioidosis is an established endemic infection throughout Sri Lanka, and is caused by multiple genotypes of B. pseudomallei, which form a distinct geographic group based upon related sequence types (BURST) cluster at the junction of the southeast Asian and Australasian clades.

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“…In screening analysis of microbial cells the use of these two matrices is common [23][24][25][26][27][28][29][30][31][32]. However, the spectra recorded with different matrices had different distribution of peaks ( Figure 3B, 3C).…”

Section: Spectrometric Evaluation Of Environmental Conditionsmentioning

confidence: 99%

Evaluation of Intact Cell Matrix-Assisted Laser Desorption/Ionization Timeof- Flight Mass Spectrometry for Capillary Electrophoresis Detection of Controlled Bacterial Clumping

Pomastowski¹,

Szultka²,

Kupczyk³

et al. 2015

J Anal Bioanal Tech

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Use of Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Analysis for Rapid Confirmation of Burkholderia pseudomallei in Septicemic Melioidosis

Cited by 35 publications

References 12 publications

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

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

Sri Lankan National Melioidosis Surveillance Program Uncovers a Nationwide Distribution of Invasive Melioidosis

Evaluation of Intact Cell Matrix-Assisted Laser Desorption/Ionization Timeof- Flight Mass Spectrometry for Capillary Electrophoresis Detection of Controlled Bacterial Clumping

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