Ikeda Masafumi, Yamaguchi Nobuyasu, Tani Katsuji, Nasu Masao
Graduate School of Pharmaceutical Sciences, Osaka University, 1-6, Yamada-oka, Suita, Osaka 565-0871, Japan.
J Microbiol Methods. 2006 Nov;67(2):241-7. doi: 10.1016/j.mimet.2006.03.014. Epub 2006 May 5.
A rapid bead assay for detecting pathogenic bacteria with a simple microfluidic chip-based system was developed. Five oligonucleotide probes corresponding to the 16S rRNA of the targeted bacteria were coupled covalently to fluorescent beads. Four species of bacteria (Escherichia coli, Salmonella enterica subsp. enterica serovar Enteritidis, Yersinia enterocolitica, and Bacillus cereus) were used as representative food-borne pathogenic bacteria. The RNAs extracted from pure cultures of these microorganisms were fluorescently labeled and hybridized to the oligonucleotide probes-immobilized fluorescent beads (Bead assay). The duplexes of RNAs and the probes-immobilized beads were analyzed with the commercially available microfluidic chip-based system. This bead assay provided results within 3 h following RNA extraction from bacterial cells.
开发了一种基于简单微流控芯片系统检测病原菌的快速磁珠检测法。将与目标细菌16S rRNA对应的五种寡核苷酸探针共价偶联到荧光磁珠上。使用四种细菌（大肠杆菌、肠炎沙门氏菌肠炎亚种肠炎血清型、小肠结肠炎耶尔森氏菌和蜡样芽孢杆菌）作为代表性食源病原菌。从这些微生物的纯培养物中提取的RNA进行荧光标记，并与固定有寡核苷酸探针的荧光磁珠杂交（磁珠检测法）。用市售的基于微流控芯片的系统分析RNA与固定有探针的磁珠的双链体。这种磁珠检测法在从细菌细胞中提取RNA后的3小时内提供结果。
