Determination of Bacterial Protein Profiles by Matrix-assisted Laser Desorption/Ionization Mass Spectrometry with High-performance Liquid Chromatography

Liang, Xiaoli; Zheng, Kefei; Qian, Masrk G.; Lubman, David M.

doi:10.1002/(sici)1097-0231(19960731)10:10<1219::aid-rcm660>3.0.co;2-r

Cited by 72 publications

(56 citation statements)

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“…In the past decade, extraction of bacterial proteins for sequencing using tandem MS/MS (8 -11) or for proteome profiling followed by matching MS spectrum results to databases (fingerprinting) have been used for bacterial identification (6,8,(12)(13)(14)(15). Despite much improvement (10, 11, 16 -20), neither de novo amino acid sequencing nor protein fingerprinting has been applied to clinical or epidemiologic uses because they are relatively time-consuming and technique-demanding or have low reproducibility (16,19).…”

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confidence: 99%

Highly Efficient Classification and Identification of Human Pathogenic Bacteria by MALDI-TOF MS

Hsieh

Tseng

Lee

et al. 2008

Molecular & Cellular Proteomics

165

111

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Accurate and rapid identification of pathogenic microorganisms is of critical importance in disease treatment and public health. Conventional work flows are time-consuming, and procedures are multifaceted. MS can be an alternative but is limited by low efficiency for amino acid sequencing as well as low reproducibility for spectrum fingerprinting. We systematically analyzed the feasibility of applying MS for rapid and accurate bacterial identification. Directly applying bacterial colonies without further protein extraction to MALDI-TOF MS analysis revealed rich peak contents and high reproducibility. The MS spectra derived from 57 isolates comprising six human pathogenic bacterial species were analyzed using both unsupervised hierarchical clustering and supervised model construction via the Genetic Algorithm. Hierarchical clustering analysis categorized the spectra into six groups precisely corresponding to the six bacterial species. Precise classification was also maintained in an independently prepared set of bacteria even when the numbers of m/z values were reduced to six. In parallel, classification models were constructed via Genetic Algorithm analysis. A model containing 18 m/z values accurately classified independently prepared bacteria and identified those species originally not used for model construction. Moreover bacteria fewer than 10 4 cells and different species in bacterial mixtures were identified using the classification model approach. In conclusion, the application of MALDI-TOF MS in combination with a suitable model construction provides a highly accurate method for bacterial classification and identification. The approach can identify bacteria with low abundance even in mixed flora, suggesting that a rapid and accurate bacterial identification using MS techniques even before culture can be attained in the near future. Molecular & Cellular Proteomics 7: 448 -456, 2008.Currently the most popular methods for bacterial identification are based on microbiologic procedures, antibody recognition, and PCR amplification. Traditionally microbiologic methods are culture-based assays that examine the presence of bacterial species. These methods provide high sensitivity and specificity, but their efficiency is limited by the complexity of the procedures, including culture, selection, isolation, and morphologic and biochemical characterization, which usually take 48 h or longer. Serologic methods are presumptive and confined to the availability of antibodies and to bacteria that are included ahead in the assays. Molecular biology techniques, particularly PCR, have been regarded as non-culturebased methods with high efficiency and specificity (1). However, they are completely dependent on the known genetic sequences of the target bacteria.MS with its capability of de novo protein/peptide sequencing (such as electrospray ionization or MALDI-TOF MS for tandem MS/MS) or its high efficiency for proteome profiling (particularly MALDI-TOF MS) has been suggested as an alternative for microbial identification (2-7...

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confidence: 99%

Highly Efficient Classification and Identification of Human Pathogenic Bacteria by MALDI-TOF MS

Hsieh

Tseng

Lee

et al. 2008

Molecular & Cellular Proteomics

165

111

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“…The analysis of intact bacteria has thus far been dominated by matrix-assisted laser desorption-ionization (MALDI)-MS (1,7,9,10,19,21,23). Protein biomarkers have been shown to be readily accessible with MALDI (1,8,24), and computer programs are under development to identify the microorganisms in the sample by searching libraries of reference spectra (18) and by searching genome or proteome databases (phyloproteomics) (9; F. Pineda, J. Linn, C. Fenselau, and P. A. Demirev, submitted for publication).…”

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confidence: 99%

Rapid Characterization of Spores of Bacillus cereus Group Bacteria by Matrix-Assisted Laser Desorption-Ionization Time-of-Flight Mass Spectrometry

Ryzhov

Hathout

Fenselau

2000

Appl Environ Microbiol

137

110

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Matrix-assisted laser desorption-ionization (MALDI) time-of-flight mass spectrometry was used to characterize the spores of 14 microorganisms of the Bacillus cereus group. This group includes the four Bacillus species B. anthracis, B. cereus, B. mycoides, and B. thuringiensis. MALDI mass spectra obtained from whole bacterial spores showed many similarities between the species, except for B. mycoides. At the same time, unique mass spectra could be obtained for the different B. cereus and B. thuringiensis strains, allowing for differentiation at the strain level. To increase the number of detectable biomarkers in the usually peak-poor MALDI spectra of spores, the spores were treated by corona plasma discharge (CPD) or sonicated prior to MALDI analysis. Spectra of sonicated or CPD-treated spores displayed an ensemble of biomarkers common for B. cereus group bacteria. Based on the spectra available, these biomarkers differentiate B. cereus group spores from those of Bacillus subtilis and Bacillus globigii. The effect of growth medium on MALDI spectra of spores was also explored.

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“…As concerns the MS detection of biomarkers, the matter is more complicated. SDS PAGE shows the presence of compounds with an MW well above that observed with MALDI in a comparative analysis of the same bacteria lysates (see for example [81,83]). Discrimination effects in MALDI are likely to result from variation in the ionisation efficiency, which, in turn, depends on experimental conditions (e.g.…”

Section: 63mentioning

confidence: 94%

“…The authors note that reproducibility, speed and sensitivity of analysis are major advantages of MALDI MS over SDS-PAGE. After this initial study, the characterisation method was extended with a capillary micro-LC separation between cell lysis and MALDI MS analysis [81]. The LC fractions obtained after gradient elution were mixed with matrix solution and deposited on a MALDI plate or on nitrocellulose substrate; the substrate reportedly improves ion yields [82].…”

Section: Analysis Of Proteinsmentioning

confidence: 99%

Characterisation of bacteria by matrix-assisted laser desorption/ionisation and electrospray mass spectrometry

Baar¹

2000

FEMS Microbiol Rev

176

143

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Determination of Bacterial Protein Profiles by Matrix-assisted Laser Desorption/Ionization Mass Spectrometry with High-performance Liquid Chromatography

Cited by 72 publications

References 0 publications

Highly Efficient Classification and Identification of Human Pathogenic Bacteria by MALDI-TOF MS

Highly Efficient Classification and Identification of Human Pathogenic Bacteria by MALDI-TOF MS

Rapid Characterization of Spores of Bacillus cereus Group Bacteria by Matrix-Assisted Laser Desorption-Ionization Time-of-Flight Mass Spectrometry

Characterisation of bacteria by matrix-assisted laser desorption/ionisation and electrospray mass spectrometry

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