MALDI-TOF MS-based analysis of dried seed proteins immobilized on filter paper

Abstract Matrix-assisted laser-desorption and ionization time-of-flight mass spectroscopy (MALDI-TOF MS) is commonly used for the characterization of protein-containing biological samples. For this, we have previously developed sample-preparation methods that can be used for discrimination between Impatiens species and also between regional biotypes of Himalayan balsam (Impatiens glandulifera), initially using leaf samples and, more recently, using seed material. In the current article, we have developed a further MALDI-TOF MS-based method that can be used with seeds that uses only simple equipment and minimally hazardous reagents prior to storing and/or shipping dried seed proteins immobilized on filter paper for MALDI-TOF MS analysis. We have investigated I. glandulifera regional-biotype seeds originating from four different sites within the UK for which the parent plants differ in their susceptibility to the biological control agent Puccinia komarovii var. glanduliferae. Using a combination of time-course comparisons and principal-component analysis, we have demonstrated good MALDI-TOF MS spectral conservation, even after storage for 1 month at 35°C, of dried seed-protein samples immobilized on filter paper. This method may provide a further useful tool for the matching of biological control agents optimally to susceptible (regional) target-plant biotypes, and for seed characterization and/or identification in general.

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MALDI-TOF MS-Based Analysis of Seed Proteins from Catalogue Varieties of Solanum lycopersicum/Lycopersicon esculentum

Horticulturae ◽

10.3390/horticulturae5030048 ◽

2019 ◽

Vol 5 (3) ◽

pp. 48

Author(s):

Michael Reeve

Keyword(s):

Lycopersicon Esculentum ◽

Sample Preparation ◽

Solanum Lycopersicum ◽

Seed Proteins ◽

Impatiens Glandulifera ◽

Invasive Weed ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Commercial Plant ◽

Tof Ms

Matrix-assisted laser-desorption and ionization time-of-flight mass spectroscopy (MALDI-TOF MS) is a flexible technique for the analysis of protein-containing biological samples. Simple and inexpensive methods have previously been developed for MALDI-TOF MS sample preparation that are able to discriminate between Impatiens species that are closely related and also between regional biotypes of the invasive weed Impatiens glandulifera (Himalayan balsam) with leaf material and also seed material. The current article investigates whether MALDI-TOF MS, through acid-soluble protein ‘fingerprinting’, can be used to analyze plant seeds that result from intensive commercial plant-breeding activity. As an initial proof-of-concept study, tomato seeds from eleven seed-catalogue varieties (F1 Pink Baby Plum, F1 Fantasio, F1 Lizzano, F1 Sungold, F1 Tumbler, Faworyt, Golden Sunrise, Hundreds and Thousands, Indigo Rose, Moneymaker, and Red Alert), listed as Solanum lycopersicum or under the synonym Lycopersicon esculentum were analyzed using MALDI-TOF MS. Whilst peak-rich and highly-reproducible spectra were obtained, with very high Bruker comparison scores and low MALDI-TOF MS variance, sample-preparation variance, and seed-to-seed variance, the spectral differences between varieties were only slightly greater than the above combined variances, indicating very close similarity between all eleven varieties studied. These results are discussed in comparison with those previously observed with the naturally-evolving invasive species I. glandulifera.

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Discrimination between Asian populations of the parasitoid wasp Ganaspis cf. brasiliensis using a simple MALDI-TOF MS-based method for use with insects

Biology Methods and Protocols ◽

10.1093/biomethods/bpz002 ◽

2019 ◽

Vol 4 (1) ◽

Cited By ~ 3

Author(s):

Michael A Reeve ◽

M Lukas Seehausen

Keyword(s):

Reference Sample ◽

Principal Component ◽

Parasitoid Wasp ◽

Test Sample ◽

Invasive Pest ◽

Drosophila Suzukii ◽

Maldi Tof Ms ◽

Chinese Populations ◽

Maldi Tof ◽

Tof Ms

Abstract The fruit fly Drosophila suzukii has recently become an invasive pest insect of significant economic impact in Europe and the USA. In contrast to other Drosophila species, D. suzukii is able to infest intact fruit by means of a saw-like ovipositor, which allows females to deposit eggs beneath the skin of the fruit. Classical biological control using the parasitoid wasp Ganaspis cf. brasiliensis is currently being researched as an environmentally sustainable option for the control of D. suzukii. In particular, the host specificity of this parasitoid has been assessed for populations from different regions in China and Japan. In order to study the relationship between the differences in specificity and molecular variations, we have adapted a matrix-assisted laser-desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based method, originally developed for use with plant material, to discriminate between example populations of G. cf. brasiliensis. We have employed a combination of principal component analysis and blind-tested comparison between reference sample MALDI-TOF MS spectra and test sample spectra to discriminate, on the basis of the acid-soluble insect protein spectra generated, between four populations of G. cf. brasiliensis (originally collected from Tokyo and Hasuike in Japan and Dali and Ximing in China). MALDI-TOF MS analysis is able to discriminate with 100% accuracy between populations G. cf. brasiliensis. The Chinese populations were observed to be similar, but the Tokyo population is slightly different and the Hasuike population is significantly different from the other populations. The Tokyo population appears more closely related to the Chinese populations than the Hasuike population, even though both originate from Japan.

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Novel Method for Reliable Identification of Siccibacter and Franconibacter Strains: from “Pseudo-Cronobacter” to New Enterobacteriaceae Genera

Applied and Environmental Microbiology ◽

10.1128/aem.00234-17 ◽

2017 ◽

Vol 83 (13) ◽

Cited By ~ 6

Author(s):

Barbora Svobodová ◽

Jiří Vlach ◽

Petra Junková ◽

Ludmila Karamonová ◽

Martina Blažková ◽

...

Keyword(s):

Intact Cell ◽

Foodborne Pathogen ◽

Principal Component ◽

Powdered Infant Formula ◽

Content Type ◽

Reliable Identification ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Maldi Biotyper ◽

Tof Ms

ABSTRACT In the last decade, strains of the genera Franconibacter and Siccibacter have been misclassified as first Enterobacter and later Cronobacter. Because Cronobacter is a serious foodborne pathogen that affects premature neonates and elderly individuals, such misidentification may not only falsify epidemiological statistics but also lead to tests of powdered infant formula or other foods giving false results. Currently, the main ways of identifying Franconibacter and Siccibacter strains are by biochemical testing or by sequencing of the fusA gene as part of Cronobacter multilocus sequence typing (MLST), but in relation to these strains the former is generally highly difficult and unreliable while the latter remains expensive. To address this, we developed a fast, simple, and most importantly, reliable method for Franconibacter and Siccibacter identification based on intact-cell matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS). Our method integrates the following steps: data preprocessing using mMass software; principal-component analysis (PCA) for the selection of mass spectrum fingerprints of Franconibacter and Siccibacter strains; optimization of the Biotyper database settings for the creation of main spectrum projections (MSPs). This methodology enabled us to create an in-house MALDI MS database that extends the current MALDI Biotyper database by including Franconibacter and Siccibacter strains. Finally, we verified our approach using seven previously unclassified strains, all of which were correctly identified, thereby validating our method. IMPORTANCE We show that the majority of methods currently used for the identification of Franconibacter and Siccibacter bacteria are not able to properly distinguish these strains from those of Cronobacter. While sequencing of the fusA gene as part of Cronobacter MLST remains the most reliable such method, it is highly expensive and time-consuming. Here, we demonstrate a cost-effective and reliable alternative that correctly distinguishes between Franconibacter, Siccibacter, and Cronobacter bacteria and identifies Franconibacter and Siccibacter at the species level. Using intact-cell MALDI-TOF MS, we extend the current MALDI Biotyper database with 11 Franconibacter and Siccibacter MSPs. In addition, the use of our approach is likely to lead to a more reliable identification scheme for Franconibacter and Siccibacter strains and, consequently, a more trustworthy epidemiological picture of their involvement in disease.

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Principal component analysis of MALDI-TOF MS of whole-cell foodborne pathogenic bacteria

Analytical Biochemistry ◽

10.1016/j.ab.2020.113582 ◽

2020 ◽

Vol 592 ◽

pp. 113582 ◽

Cited By ~ 2

Author(s):

Wenjing Yan ◽

Jing Qian ◽

Yongjie Ge ◽

Keping Ye ◽

Cunshan Zhou ◽

...

Keyword(s):

Principal Component Analysis ◽

Pathogenic Bacteria ◽

Principal Component ◽

Component Analysis ◽

Whole Cell ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Foodborne Pathogenic Bacteria ◽

Tof Ms

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Evaluation of filter paper as a means to transport inactivated bacteria for identification using the MALDI-TOF MS system

Journal of Microbiological Methods ◽

10.1016/j.mimet.2020.105863 ◽

2020 ◽

Vol 171 ◽

pp. 105863 ◽

Cited By ~ 1

Author(s):

M.S. Carneiro ◽

A. Fracasso ◽

O.V.A. Lovison ◽

F. Barreto ◽

A.L. Barth

Keyword(s):

Filter Paper ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Tof Ms

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Collagen-based proteinaceous binder-pigment interaction study under UV ageing conditions by MALDI-TOF-MS and principal component analysis

Journal of Mass Spectrometry ◽

10.1002/jms.2966 ◽

2012 ◽

Vol 47 (3) ◽

pp. 322-330 ◽

Cited By ~ 17

Author(s):

Julia Romero-Pastor ◽

Natalia Navas ◽

Stepanka Kuckova ◽

Alejandro Rodríguez-Navarro ◽

Carolina Cardell

Keyword(s):

Principal Component Analysis ◽

Principal Component ◽

Component Analysis ◽

Interaction Study ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Uv Ageing ◽

Proteinaceous Binder ◽

Tof Ms

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Differentiation between closely-related Impatiens spp. and regional biotypes of Impatiens glandulifera using a highly-simplified and inexpensive method for MALDI-TOF MS

Plant Methods ◽

10.1186/s13007-018-0323-6 ◽

2018 ◽

Vol 14 (1) ◽

Cited By ~ 10

Author(s):

Michael A. Reeve ◽

Kathryn M. Pollard ◽

Daisuke Kurose

Keyword(s):

Inexpensive Method ◽

Impatiens Glandulifera ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Tof Ms

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Discrimination of Bacillus cereus Group Members by MALDI-TOF Mass Spectrometry

Microorganisms ◽

10.3390/microorganisms9061202 ◽

2021 ◽

Vol 9 (6) ◽

pp. 1202

Author(s):

Viviana Manzulli ◽

Valeria Rondinone ◽

Alessandro Buchicchio ◽

Luigina Serrecchia ◽

Dora Cipolletta ◽

...

Keyword(s):

Mass Spectrometry ◽

Bacillus Cereus ◽

Mass Spectra ◽

Critical Time ◽

Principal Component ◽

Etiologic Agent ◽

Bacillus Cereus Group ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Tof Ms

Matrix-Assisted Laser Desorption/Ionization Time Of Flight Mass Spectrometry (MALDI-TOF MS) technology is currently increasingly used in diagnostic laboratories as a cost effective, rapid and reliable routine technique for the identification and typing of microorganisms. In this study, we used MALDI-TOF MS to analyze a collection of 160 strains belonging to the Bacillus cereus group (57 B. anthracis, 49 B. cereus, 1 B. mycoides, 18 B. wiedmannii, 27 B. thuringiensis, 7 B. toyonensis and 1 B. weihenstephanensis) and to detect specific biomarkers which would allow an unequivocal identification. The Main Spectra Profiles (MSPs) were added to an in-house reference library, expanding the current commercial library which does not include B. toyonensis and B. wiedmannii mass spectra. The obtained mass spectra were statistically compared by Principal Component Analysis (PCA) that revealed seven different clusters. Moreover, for the identification purpose, were generated dedicate algorithms for a rapid and automatic detection of characteristic ion peaks after the mass spectra acquisition. The presence of specific biomarkers can be used to differentiate strains within the B. cereus group and to make a reliable identification of Bacillus anthracis, etiologic agent of anthrax, which is the most pathogenic and feared bacterium of the group. This could offer a critical time advantage for the diagnosis and for the clinical management of human anthrax even in case of bioterror attacks.

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