Detection of Escherichia coli in Meat with an Electrochemical Biochip
Detection of Escherichia coli in Meat with an Electrochemical Biochip
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DOI：
10.4315/0362-028x-73.11.2025
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发表时间：
2010-11-01
影响因子：
2
通讯作者：
Gareis, Manfred
中科院分区：
文献类型：
--
作者：
Heidenreich, Bernd;Poehlmann, Christopher;Gareis, Manfred
通过RNA-DNA杂交检测食源性致病菌和腐败菌是传统微生物方法的替代方法。为了用基于RNA-DNA的方法实现高灵敏度，需要有效的细菌裂解和从细菌释放核酸。在这里，我们报告的卫生指示微生物大肠杆菌的特异性检测肉类使用的电化学生物芯片。我们改进了从猪肉和牛肉的肉汁中提取细菌RNA的方法。对自然或人工污染的样品进行了E.杆菌通过在完全培养基或基本培养基中孵育来富集。用溶菌酶、蛋白酶K联合处理样品。超声处理导致有效的细胞破碎和高的总RNA产量。与富集时间的优化一起，这确保了生物芯片上电化学测量的高灵敏度。A.短富集期和三次裂解方案结合电化学生物芯片测量对25个肉样品进行了测试。生物芯片的检测下限为约2，000 CFU的E. coli/ml。整个分析程序（5小时富集、三重裂解和生物芯片检测）的检测下限为1 CFU E。大肠杆菌/ml。
Detection of foodborne pathogenic and spoilage bacteria by RNA-DNA hybridization is an alternative to traditional microbiological procedures. To achieve high sensitivity with RNA-DNA-based methods, efficient bacterial lysis and release of nucleic acids from bacteria are needed. Here we report the specific detection of the hygiene indicator microorganism Escherichia coli in meat by use of electrochemical biochips. We improved RNA isolation from bacteria in meat juice from pork and beef. Samples, either naturally or artificially contaminated by E. coli. were enriched by incubation in full or minimal medium. A combined treatment of the samples with lysozyme, proteinase K. and sonication resulted in efficient cell disruption and high total RNA yields. Together with optimization of enrichment time, this ensures high sensitivity of electrochemical measurements on biochips. A. short enrichment period and the triple-lysis regimen in combination with electrochemical biochip measurement were tested with 25 meat samples. The lower limit of detection of the biochip was approximately 2,000 CFU of E. coli per ml. The entire analysis procedure (5 h of enrichment, triple lysis, and biochip detection) has a lower limit of detection of 1 CFU of E. coli per ml within a total time needed for analysis of 7 h.