Simple Bead Assay for Detection of Live Bacteria (Escherichia coli)
Simple Bead Assay for Detection of Live Bacteria (Escherichia coli)
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DOI：
10.1021/ac103109v
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发表时间：
2011-02-15
影响因子：
7.4
通讯作者：
Goldys, Ewa M.
中科院分区：
文献类型：
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作者：
Butterworth, Philip;Baltar, Henrique T. M. C. M.;Goldys, Ewa M.
微珠检测是一种重要的快速微生物检测技术，适用于极低的病原体水平。我们报告了从大肠杆菌K12中提取的rRNA的珠测定，其不需要扩增步骤，并且在Agilent 2100 Bioanalyzer流式细胞仪系统上具有读数。我们的检测方法能够检测到125 ng的RNA，这比之前报道的少16倍。特异性极高，与阴性对照微生物（枯草芽孢杆菌）无结合。我们讨论了在优化关键检测组件过程中面临的挑战，例如样品中RNA的含量、珠粒数量、聚集和再现性。
Bead assays are an important rapid microbial detection technology suitable for extremely low pathogen levels. We report a bead assay for rRNA extracted from Escherichia coli K12 that does not require amplification steps and has readout on an Agilent 2100 Bioanalyzer flow cytometry system. Our assay was able to detect 125 ng of RNA, which is 16 times less than reported earlier. The specificity was extremely high, with no binding to a negative control organism (Bacillus subtilis). We discuss challenges faced during optimization of the key assay components, such as varying amounts of RNA in the samples, number of beads, aggregation, and reproducibility.