title: Comparison of different methods for the recovery of DNA from spores of mycotoxin-producing moulds in spiked food samples.
abstract: Several food samples were spiked with fungal conidia to test the efficiency of different cell disruption methods and DNA extraction kits for subsequent molecular detection. For disrupting the firm cell walls of the spores, two different pretreatment methods, namely sonication and bead beating, were tested against no pretreatment. The subsequent DNA extraction and purification was performed using three different DNA extraction methods, which are based on a diverse combination of extraction principles, such as precipitation, thermic-enzymatic lysis, pH-enhancement and bonding with a silica membrane. The aim of the study was to find out the suitable pretreatment and DNA extraction method for the recovery of detectable amounts of fungal DNA from different food matrices. Significance and impact of the study: The choice of 'ready-to-use' commercial kits and methods has been of great importance regarding the recovery of extracted DNA. However, these commercially available kits are neither effective nor time-efficient when extracting DNA from fungal spores embedded in complex food matrices. Different extraction principles were compared and their effectiveness tested using real-time PCR. The combination of different principles for the extraction and purification of DNA was found as the most efficient method (quantity and purity) to obtain DNA from moulds and their spores from food samples.
authors.affiliation: Microbiology and Food Hygiene, Niederrhein University of Applied Sciences, Mönchengladbach, Germany.|Institute of Microbiology and Virology, University of Witten/Herdecke, Witten, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Grube
authors.affiliation: GEN-IAL GmbH, Troisdorf, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Schönling
authors.affiliation: Microbiology and Food Hygiene, Niederrhein University of Applied Sciences, Mönchengladbach, Germany.|Institute of Microbiology and Virology, University of Witten/Herdecke, Witten, Germany.|Center for Advanced Microstructures and Devices (CAMD), Louisiana State University, Baton Rouge, LA, USA.
authors.forename: A
authors.initials: A
authors.lastname: Prange
mesh_terms: D004271:DNA, Fungal / Q000235:genetics* / Q000302:isolation & purification; D005516:Food Microbiology / Q000379:methods*; D005658:Fungi / Q000235:genetics*; D006801:Humans; D009183:Mycotoxins / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D013172:Spores, Fungal / Q000737:chemistry
chemical_list: D004271:DNA, Fungal; D009183:Mycotoxins
keywords: DNA extraction; cell disruption; food quality assurance; food safety; moulds; real-time PCR; spores