TY - Generic T1 - Proteomic detection as an alternative for the quantification of Staphylococcus aureus enterotoxins Y1 - 2014 A1 - Mirjana Andjelkovic A1 - V. Tsilia A1 - A. Rajkovic A1 - S. Cosijns A1 - Koen De Cremer A1 - Joris Van Loco KW - additional KW - alternative KW - an KW - analysi KW - analysis KW - approach KW - approaches KW - AS KW - at KW - bacteria KW - cause KW - Common KW - Control KW - detection KW - Development KW - disease KW - Diseases KW - Efficiency KW - enterotoxin KW - Enterotoxins KW - environment KW - estimation KW - food KW - Foodborne Diseases KW - foodborne outbreaks KW - Foods KW - Gastroenteritis KW - identification KW - identify KW - Immunoassay KW - IS KW - matrix KW - Meat KW - method KW - methods KW - microorganism KW - milk KW - ON KW - outbreak KW - Peptides KW - poisoning KW - prevention KW - PROCESSES KW - protein KW - purification KW - Quantification KW - RANGE KW - Sample KW - Samples KW - Selection KW - specific KW - staphylococcus KW - Staphylococcus aureus KW - structure AB -

Food poisoning caused by ingestion of Staphylococcus aureus enterotoxins is one of the most common foodborne diseases. Staphylococcus aureus is a well-studied, omnipresent bacterium which is not only found in the environment but is also part of the commensal mammalian flora. This microorganism produces enterotoxins, protein by their structure which can cause gastro-enteritis, emesis or act as superantigen. The methods used to its confirmation in food samples are mainly of microbiological character and are not quantitative. They may not be used in prevention of any foodborne outbreaks or to properly identify them. Besides, none of those methods allow unambiguous identification, nor quantification, as molecular tools are inefficient at proving the existence of the toxins in foods and immunoassays are not specific enough, not suitable for quantification and more importantly limited in the range of toxins they can identify. The proteomic approach method for the specific detection and quantification of each toxin is achieved through the analysis of peptides (toxin fragments) unique to a toxin. The peptides are obtained by extraction, purification and concentration of the enterotoxins out of the matrix and submitted to a proteotypic digestion by trypsin, The goal was to select two specific peptides per toxin to ensure proper identification and quantification. With a recognizable extraction concept the toxin is isolated from the incriminated matrix. During the process of method development the toxin was spiked at various steps down the extraction procedure to a matrix for additional control of the extraction losses. The spiked amount of toxins (1000 ng of each enterotoxin) is not representative of a real contamination but was used for the estimation. The major conclusions were that the extraction efficiency was dependent (eg milk vs meat) on the food matrix and the selection of the proteotypicpeptides

JF - ProteoMMX³ strictly quantitative T3 - ProteoMMX³ strictly quantitative - Book of Abstracts PB - NA CY - NA CP - University of Liverpool in association with the British Society for Proteome Research U1 - 2237 U2 - March 24-27th 2014 ER -