Elaboration and validation of the method for the quantification of the emetic toxin of Bacillus cereus as described in EN-ISO 18465 - Microbiology of the food chain – Quantitative determination of emetic toxin (cereulide) using LC-MS/MS

Veld, P.H. in 't; Laak, L.F.J. van der; Zon, Meng; Biesta-Peters, Elisabeth G.

doi:10.1016/j.ijfoodmicro.2018.03.021

Cited by 21 publications

(12 citation statements)

References 15 publications

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“…Accredited laboratories and food companies worldwide use these standards to ensure the safety of food products for the consumers. In order for a new method to be included in the standards catalog, it needs to undergo a validation procedure that compares the performances of the new method to the reference one (In ‘t Veld, van der Laak, van Zon, & Biesta‐Peters, 2019). The Comité European de Normalization (CEN) has developed a standard procedure for the validation of alternative methods in food microbiology, EN ISO 16140 (Anonymous, 2003).…”

Section: Introductionmentioning

confidence: 99%

Microbial detection and identification methods: Bench top assays to omics approaches

Ferone

Gowen

Fanning

et al. 2020

Comp Rev Food Sci Food Safe

146

111

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Rapid detection of foodborne pathogens, spoilage microbes, and other biological contaminants in complex food matrices is essential to maintain food quality and ensure consumer safety. Traditional methods involve culturing microbes using a range of nonselective and selective enrichment methods, followed by biochemical confirmation among others. The time‐to‐detection is a key limitation when testing foods, particularly those with short shelf lives, such as fresh meat, fish, dairy products, and vegetables. Some recent detection methods developed include the use of spectroscopic techniques, such as matrix‐assisted laser desorption ionization‐time of flight along with hyperspectral imaging protocols.This review presents a comprehensive overview comparing insights into the principles, characteristics, and applications of newer and emerging techniques methods applied to the detection and identification of microbes in food matrices, to more traditional benchtop approaches. The content has been developed to provide specialist scientists a broad view of bacterial identification methods available in terms of their benefits and limitations, which may be useful in the development of future experimental design. The case is also made for incorporating some of these emerging methods into the mainstream, for example, underutilized potential of spectroscopic techniques and hyperspectral imaging.

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Section: Introductionmentioning

confidence: 99%

Microbial detection and identification methods: Bench top assays to omics approaches

Ferone

Gowen

Fanning

et al. 2020

Comp Rev Food Sci Food Safe

146

111

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“…Thus, liquid chromatography coupled to mass spectrometry (LC-MS) is still considered the gold standard for cereulide detection and quantification [33][34][35][36]. Based on stable isotope mass spectrometry-based dilution assay (MS-SIDA) [33], an EN-ISO method (EN ISO 18465) for quantitation of cereulide in food matrices was recently established [37].…”

Section: Introductionmentioning

confidence: 99%

Detection and Isolation of Emetic Bacillus cereus Toxin Cereulide by Reversed Phase Chromatography

Kalbhenn

Bauer

Stark

et al. 2021

Toxins

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The emetic toxin cereulide is a 1.2 kDa dodecadepsipeptide produced by the food pathogen Bacillus cereus. As cereulide poses a serious health risk to humans, sensitive and specific detection, as well as toxin purification and quantification, methods are of utmost importance. Recently, a stable isotope dilution assay tandem mass spectrometry (SIDA–MS/MS)-based method has been described, and an method for the quantitation of cereulide in foods was established by the International Organization for Standardization (ISO). However, although this SIDA–MS/MS method is highly accurate, the sophisticated high-end MS equipment required for such measurements limits the method’s suitability for microbiological and molecular research. Thus, we aimed to develop a method for cereulide toxin detection and isolation using equipment commonly available in microbiological and biochemical research laboratories. Reproducible detection and relative quantification of cereulide was achieved, employing reversed phase chromatography (RPC). Chromatographic signals were cross validated by ultraperformance liquid chromatography–mass spectrometry (UPLC–MS/MS). The specificity of the RPC method was tested using a test panel of strains that included non-emetic representatives of the B. cereus group, emetic B. cereus strains, and cereulide-deficient isogenic mutants. In summary, the new method represents a robust, economical, and easily accessible research tool that complements existing diagnostics for the detection and quantification of cereulide.

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“…The direct identification of toxins in food can be carried out by LC-MS/MS or HPLC MS. These methods are able to identify and quantify the toxin with high a sensitivity (10, 13, 14). Generally, food matrices rich in carbohydrates, such as pasta and rice, as well as milk and dairy products have the highest risk of causing cereulide intoxications (6, 15).…”

Section: Introductionmentioning

confidence: 99%

Identification of cereulide producingBacillus cereusby MALDI-TOF MS

Ulrich

Gottschalk

Dietrich

et al. 2018

Preprint

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26The Bacillus (B.) cereus group is genetically highly homogenous and consists of nine recognized 27 species which are present worldwide. B. cereus sensu stricto play an important role in food-borne 28 diseases by producing different toxins. Yet, only a small percentage of B. cereus strains are able to 29 produce the heat stable depsipeptide cereulide, the causative agent of emetic food poisonings. To 30 minimize the entry of emetic B. cereus into the food chain, food business operators are dependent on 31 efficient and reliable methods enabling the differentiation between emetic and non-emetic strains. 32Currently, only time-consuming cell bioassays, molecular methods and tandem mass spectrometry are 33 available for this purpose. Thus, the aim of the present study was to establish a fast and reliable 34 method for the differentiation between emetic and non-emetic strains by MALDI-TOF MS. Selected 35 isolates/strains of the B. cereus group (total n=110, i.e. emetic n=45, non-emetic n=65) were cultured 36 on sheep blood agar for 48h.37 Subsequently, the cultures were directly analyzed by MALDI-TOF MS without prior extraction steps 38 (direct smear method). The samples were measured in linear positive ionization mode in the mass 39 range of m/z 800 -1,800 Da. Using ClinProTools 3.0 statistical software and flex analyst, a 40 differentiation between emetic and non-emetic isolates was possible with a rate of correct 41 identification of 99.1 % by means of the evaluation of two specific biomarkers (m/z 1171 and 1187 42 Da).43 44 Importance 45 Bacillus (B.) cereus plays an important role in food-borne diseases due to the production of different 46 toxins, e.g. the heat stable depsipeptide cereulide. Only a small number of B. cereus strains are able to 47 produce this toxin, the causative agent of emetic food poisonings. To minimize the entry of emetic B. 48 cereus into the food chain, food business operators require efficient and reliable methods enabling the 49 differentiation between emetic and non-emetic strains. The aim of the present study was to develop a50 fast and reliable method for the differentiation between emetic and non-emetic strains by MALDI-51 TOF MS. A differentiation between emetic and non-emetic isolates was possible with a rate of correct identification of 99.1 % by means of the evaluation of two specific biomarkers (m/z 1171 and 1187 53 Da). 54 55 57 58 Bacillus (B.) spp. are Gram-positive, rod shaped bacteria occurring world-wide. The B. cereus group is 59 genetically highly homogeneous and comprises nine recognized species: B. anthracis, B. cereus sensu 60 stricto, B. cytotoxicus, B. mycoides, B. pseudomycoides, B. thuringiensis, B. toyonensis, B. 61 weihenstephanensis and B. wiedmannii (1, 2). Their spores are heat, acid, UV and desiccation resistant 62 and survive pasteurization (3, 4). 63Due to the persistence of spores in food and the ability of the vegetative cells to produce different 64 toxins, isolates of the B. cereus group play an important role in food safety. Several publications...

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Elaboration and validation of the method for the quantification of the emetic toxin of Bacillus cereus as described in EN-ISO 18465 - Microbiology of the food chain – Quantitative determination of emetic toxin (cereulide) using LC-MS/MS

Cited by 21 publications

References 15 publications

Microbial detection and identification methods: Bench top assays to omics approaches

Microbial detection and identification methods: Bench top assays to omics approaches

Detection and Isolation of Emetic Bacillus cereus Toxin Cereulide by Reversed Phase Chromatography

Identification of cereulide producingBacillus cereusby MALDI-TOF MS

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