中文摘要：本文开发了一种对香料和草本植物病原菌进行灵敏检测的方法并对其进行了评估，该方法采用十六烷基三甲基溴化铵（CTAB）提取DNA、通用细菌聚合酶链反应扩增16S rDNA、在Illumina MiSeq平台上进行高通量测序，并采用受沙门氏菌、大肠杆菌（革兰氏阴性菌为代表）和金黄色葡萄球菌（如革兰氏阳性细菌的代表）不同程度污染的样本进行灵敏度评估。检测结果显示：该方法对香辛料（辣椒，黑胡椒调料）样本检测灵敏度可达到筛选水平，检测限在10(4)-10(5) CFU/g范围内且半定量响应。对草本植物（牛至，欧芹）检测灵敏度较低，不低于10(7) CFU/g，该方法在香辛料筛选方面具有一定的潜力，但其灵敏度有待于提高。
外文摘要：A broad-range culture-independent method was developed and evaluated regarding its sensitivity of detection of pathogenic bacteria in spices and herbs, with focus on paprika powder. The method involved DNA extraction using cetyltrimethylammonium bromide (CTAB), 16S rDNA amplification using universal bacterial polymerase chain reaction, and high-throughput sequencing on Illumina MiSeq platform. The sensitivity of the method was evaluated with series of model samples contaminated at different levels with Salmonella enterica and Escherichia coli (as representatives of Gram-negative bacteria) and Staphylococcus aureus (as a representative of Gram-positive bacteria). For spices (paprika, black pepper), the method had a screening-level sensitivity with limits of detection in the range of 10(4)-10(5) CFU/g, and a semi-quantitative response. Low sensitivity (LOD >= 10(7) CFU/g) Was observed with herbs (oregano, parsley). The developed method demonstrated a good potential for microbiological screening of spices, with a prospect of further improvement of sensitivity based on progress in high-throughput sequencing technology. (C) 2016 Elsevier Ltd. All rights reserved.
外文关键词：Food safety, Staphylococcus aureus, Salmonella enterica, Escherichia coli, Paprika
作者：Plany, M.; Soltys, K.; Budis, J.; Mader, A.; Szemes, T.; Siekel, P.; Kuchta, T.
作者单位：Natl Agr & Food Ctr, Food Res Inst, Dept Microbiol Mol Biol & Biotechnol, Priemyselna 4, SK-82475 Bratislava, Slovakia.
期刊名称：FOOD CONTROL
期刊影响因子：3.496
出版年份：2017
出版刊次：6
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