Food Targeting: A Real-Time PCR Assay Targeting 16S rDNA for Direct Quantification of Alicyclobacillus spp. Spores after Aptamer-Based Enrichment.
Identifieur interne : 000645 ( Main/Exploration ); précédent : 000644; suivant : 000646Food Targeting: A Real-Time PCR Assay Targeting 16S rDNA for Direct Quantification of Alicyclobacillus spp. Spores after Aptamer-Based Enrichment.
Auteurs : Tim Hünniger [Allemagne] ; Christine Felbinger [Allemagne] ; Hauke Wessels [Allemagne] ; Sophia Mast [Allemagne] ; Antonia Hoffmann [Allemagne] ; Anna Schefer [Allemagne] ; Erwin M Rtlbauer [Allemagne] ; Angelika Paschke-Kratzin [Allemagne] ; Markus Fischer [Allemagne]Source :
- Journal of agricultural and food chemistry [ 1520-5118 ] ; 2015.
English descriptors
- KwdEn :
- Alicyclobacillus (classification), Alicyclobacillus (genetics), Alicyclobacillus (isolation & purification), Beverages (analysis), Beverages (microbiology), Citrus sinensis (chemistry), Citrus sinensis (microbiology), DNA, Bacterial (genetics), DNA, Ribosomal (genetics), Food Contamination (analysis), RNA, Ribosomal, 16S (genetics), Real-Time Polymerase Chain Reaction (methods), Spores, Bacterial (classification), Spores, Bacterial (genetics), Spores, Bacterial (isolation & purification).
- MESH :
- chemical , genetics : DNA, Bacterial, DNA, Ribosomal, RNA, Ribosomal, 16S.
- analysis : Beverages, Food Contamination.
- chemistry : Citrus sinensis.
- classification : Alicyclobacillus, Spores, Bacterial.
- genetics : Alicyclobacillus, Spores, Bacterial.
- isolation & purification : Alicyclobacillus, Spores, Bacterial.
- methods : Real-Time Polymerase Chain Reaction.
- microbiology : Beverages, Citrus sinensis.
Abstract
Spore-forming Alicyclobacillus spp. are able to form metabolites that induce even in small amounts an antiseptical or medicinal off-flavor in fruit juices. Microbial contaminations could occur by endospores, which overcame the pasteurization process. The current detection method for Alicyclobacillus spp. can take up to 1 week because of microbiological enrichment. In a previous study, DNA aptamers were selected and characterized for an aptamer-driven rapid enrichment of Alicyclobacillus spp. spores from orange juice by magnetic separation. In the present work, a direct quantification assay for Alicyclobacillus spp. spores was developed to complete the two-step approach of enrichment and detection. After mechanical treatment of the spores, the isolated DNA was quantified in a real-time PCR-assay targeting 16S rDNA. The assay was evaluated by the performance requirements of the European Network of Genetically Modified Organisms Laboratories (ENGL). Hence, the presented method is applicable for direct spore detection from orange juice in connection with an enrichment step.
DOI: 10.1021/acs.jafc.5b00874
PubMed: 25880790
Affiliations:
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uniqKey="Mast S" first="Sophia" last="Mast">Sophia Mast</name><affiliation wicri:level="1"><nlm:affiliation>‡Lehrstuhl für Hygiene und Technologie der Milch, Tierärtzliche Fakultät, Ludwig-Maximilians-Universität, Schönleutnerstraße 8/219, 85764 Oberschleißheim, Germany.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>‡Lehrstuhl für Hygiene und Technologie der Milch, Tierärtzliche Fakultät, Ludwig-Maximilians-Universität, Schönleutnerstraße 8/219, 85764 Oberschleißheim</wicri:regionArea><wicri:noRegion>85764 Oberschleißheim</wicri:noRegion><wicri:noRegion>85764 Oberschleißheim</wicri:noRegion><wicri:noRegion>85764 Oberschleißheim</wicri:noRegion></affiliation></author><author><name sortKey="Hoffmann, Antonia" sort="Hoffmann, Antonia" uniqKey="Hoffmann A" first="Antonia" last="Hoffmann">Antonia Hoffmann</name><affiliation wicri:level="1"><nlm:affiliation>†Hamburg School of Food Science, Institute of Food Chemistry, University of Hamburg, Grindelallee 117, 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uniqKey="Paschke Kratzin A" first="Angelika" last="Paschke-Kratzin">Angelika Paschke-Kratzin</name><affiliation wicri:level="1"><nlm:affiliation>†Hamburg School of Food Science, Institute of Food Chemistry, University of Hamburg, Grindelallee 117, 20146 Hamburg, Germany.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>†Hamburg School of Food Science, Institute of Food Chemistry, University of Hamburg, Grindelallee 117, 20146 Hamburg</wicri:regionArea><wicri:noRegion>20146 Hamburg</wicri:noRegion><placeName><settlement type="city">Hambourg</settlement><region type="land" nuts="2">Hambourg</region></placeName></affiliation></author><author><name sortKey="Fischer, Markus" sort="Fischer, Markus" uniqKey="Fischer M" first="Markus" last="Fischer">Markus Fischer</name><affiliation wicri:level="1"><nlm:affiliation>†Hamburg School of Food Science, Institute of Food Chemistry, University of Hamburg, Grindelallee 117, 20146 Hamburg, Germany.</nlm:affiliation><country 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(genetics)</term><term>DNA, Ribosomal (genetics)</term><term>Food Contamination (analysis)</term><term>RNA, Ribosomal, 16S (genetics)</term><term>Real-Time Polymerase Chain Reaction (methods)</term><term>Spores, Bacterial (classification)</term><term>Spores, Bacterial (genetics)</term><term>Spores, Bacterial (isolation & purification)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Bacterial</term><term>DNA, Ribosomal</term><term>RNA, Ribosomal, 16S</term></keywords><keywords scheme="MESH" qualifier="analysis" xml:lang="en"><term>Beverages</term><term>Food Contamination</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Citrus sinensis</term></keywords><keywords scheme="MESH" qualifier="classification" xml:lang="en"><term>Alicyclobacillus</term><term>Spores, Bacterial</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Alicyclobacillus</term><term>Spores, Bacterial</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Alicyclobacillus</term><term>Spores, Bacterial</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Real-Time Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Beverages</term><term>Citrus sinensis</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Spore-forming Alicyclobacillus spp. are able to form metabolites that induce even in small amounts an antiseptical or medicinal off-flavor in fruit juices. Microbial contaminations could occur by endospores, which overcame the pasteurization process. The current detection method for Alicyclobacillus spp. can take up to 1 week because of microbiological enrichment. In a previous study, DNA aptamers were selected and characterized for an aptamer-driven rapid enrichment of Alicyclobacillus spp. spores from orange juice by magnetic separation. In the present work, a direct quantification assay for Alicyclobacillus spp. spores was developed to complete the two-step approach of enrichment and detection. After mechanical treatment of the spores, the isolated DNA was quantified in a real-time PCR-assay targeting 16S rDNA. The assay was evaluated by the performance requirements of the European Network of Genetically Modified Organisms Laboratories (ENGL). Hence, the presented method is applicable for direct spore detection from orange juice in connection with an enrichment step.</div></front></TEI><affiliations><list><country><li>Allemagne</li></country><region><li>Hambourg</li></region><settlement><li>Hambourg</li></settlement></list><tree><country name="Allemagne"><region name="Hambourg"><name sortKey="Hunniger, Tim" sort="Hunniger, Tim" uniqKey="Hunniger T" first="Tim" last="Hünniger">Tim Hünniger</name></region><name sortKey="Felbinger, Christine" sort="Felbinger, Christine" uniqKey="Felbinger C" first="Christine" last="Felbinger">Christine Felbinger</name><name sortKey="Fischer, Markus" sort="Fischer, Markus" uniqKey="Fischer M" first="Markus" last="Fischer">Markus Fischer</name><name sortKey="Hoffmann, Antonia" sort="Hoffmann, Antonia" uniqKey="Hoffmann A" first="Antonia" last="Hoffmann">Antonia Hoffmann</name><name sortKey="M Rtlbauer, Erwin" sort="M Rtlbauer, Erwin" uniqKey="M Rtlbauer E" first="Erwin" last="M Rtlbauer">Erwin M Rtlbauer</name><name sortKey="Mast, Sophia" sort="Mast, Sophia" uniqKey="Mast S" first="Sophia" last="Mast">Sophia Mast</name><name sortKey="Paschke Kratzin, Angelika" sort="Paschke Kratzin, Angelika" uniqKey="Paschke Kratzin A" first="Angelika" last="Paschke-Kratzin">Angelika Paschke-Kratzin</name><name sortKey="Schefer, Anna" sort="Schefer, Anna" uniqKey="Schefer A" first="Anna" last="Schefer">Anna Schefer</name><name sortKey="Wessels, Hauke" sort="Wessels, Hauke" uniqKey="Wessels H" first="Hauke" last="Wessels">Hauke Wessels</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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