Food Targeting: A Real-Time PCR Assay Targeting 165 rDNA for Direct Quantification of Alicyclobacillus spp. Spores after Aptamer-Based Enrichment
Food Targeting: A Real-Time PCR Assay Targeting 165 rDNA for Direct Quantification of Alicyclobacillus spp. Spores after Aptamer-Based Enrichment
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DOI：
10.1021/acs.jafc.5b00874
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发表时间：
2015-05-06
影响因子：
6.1
通讯作者：
Fischer, Markus
中科院分区：
文献类型：
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作者：
Huenniger, Tim;Felbinger, Christine;Fischer, Markus
产孢脂环酸杆菌能够形成即使少量也能在果汁中引起防腐或药用异味的代谢物。微生物污染可能会发生内生孢子，这克服了巴氏杀菌过程。脂环酸芽孢杆菌的现行检测方法。由于微生物富集，可能需要长达1周。在先前的研究中，选择DNA适体并表征用于适体驱动的脂环酸芽孢杆菌属的快速富集。从橙子汁中分离孢子。在本工作中，直接定量测定脂环酸芽孢杆菌属。孢子，以完成富集和检测的两步方法。机械处理孢子后，在靶向16S rDNA的实时PCR测定中定量分离的DNA。根据欧洲转基因生物实验室网络（ENGL）的性能要求评价了该检测试剂盒。因此，所提出的方法适用于直接孢子检测与富集步骤从橙子汁。
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.