Evaluation of DNA extraction methods for PCR detection of fungal and bacterial contamination in cocoa extracts

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dc.contributor.authorMarta Tortajada
dc.contributor.authorPedro Vicente Martinez-Culebras
dc.contributor.authorVeronica Navarro
dc.contributor.authorHonorato Monzo Daniel Ramon
dc.date.accessioned2014-04-07T10:50:05Z
dc.date.available2014-04-07T10:50:05Z
dc.date.issued2009
dc.description.abstractDirect and sensitive PCR detection of contaminant microXora in cocoa extracts is aVected by the quality of the template DNA. This study compares the eYcacy of Wve diVerent commercial DNA extraction methods, selective enrichment broths and use of glycolitic enzymes to obtain quality DNA for PCR detection of both fungi and bacteria in artiWcially inoculated cocoa extract samples. PCR-based methods were applied to detect contaminant microXora in cocoa extracts using as model organisms: Aspergillus nidulans, Bacillus subtilis, Escherichia coli and Salmonella enterica. The quality of the extracted DNA was assessed in terms of PCR inhibitor content with results indicating that the HighPure PCR template (Roche) kit was the best methodology under the conditions assayed. PCR protocols using this commercial kit and a combination of glycolitic enzymes and enrichment procedures gave a detection limit of 100 conidia/g and 100 cfu/g for Wlamentous fungi and bacteria, respectively. The selected extraction and PCR procedures were also tested to assess their suitability for detecting Wlamentous fungi and bacteria on an industrial scale. They were sensitive enough to detect fungal and bacterial contaminants within the legally required limits. The results obtained with the molecular approach were in agreement with those of standard microbiological tests but require a considerably shorter analysis time. Thus, the molecular approach provides a sensitive and rapid alternative to check for microbial contamination in cocoa extracts.en_US
dc.identifier.citationEur Food Res Technol (2009) 230:79–87en_US
dc.identifier.urihttp://hdl.handle.net/123456789/300
dc.language.isoenen_US
dc.subjectAspergillusen_US
dc.subjectBacillusen_US
dc.subjectCocoaen_US
dc.subjectDNA extractionen_US
dc.subjectPCRen_US
dc.subjectSalmonellaen_US
dc.subjectE. colien_US
dc.titleEvaluation of DNA extraction methods for PCR detection of fungal and bacterial contamination in cocoa extractsen_US
dc.typeArticleen_US

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