U.S. Food and Drug Administration San Francisco District Laboratory, Alameda, CA 94502, USA.
J Microbiol Methods. 2011 Oct;87(1):105-10. doi: 10.1016/j.mimet.2011.07.019. Epub 2011 Jul 30.
Identification and serotyping of Shiga toxin-producing Escherichia coli during foodborne outbreaks can aid in matching clinical, food, and environmental isolates when trying to identify the source of illness and ultimately food contamination. Herein we describe a Luminex microbead-based suspension array to identify the O serogroup of the ten most clinically relevant STECs: O26, O45, O91, O103, O111, O113, O121, O128, O145, and O157. The use of PCR followed by Luminex xMAP® technology enables the detection of multiple analytes in a single multiplex reaction with high throughput capabilities. One hundred and fourteen STEC isolates were correctly identified with no false positives among forty-six other organisms using this assay. Assay performance was tested in multiple laboratories using a panel of eleven different STEC serogroups on the Bio-Plex 200 and MAGPIX instruments. The STEC microbead-based suspension array can be performed in a 96-well plate format for high throughput screening in less than 4h. Furthermore, it is expandable, allowing for the addition of O serogroups should the need arise.
在食源性疾病暴发期间，对产志贺毒素大肠杆菌（Shiga toxin-producing Escherichia coli，STEC）进行鉴定和血清型分型有助于在尝试确定疾病来源和最终食物污染时，对临床、食物和环境分离株进行匹配。在此，我们描述了一种基于 Luminex 微球的悬浮阵列，用于鉴定十种最具临床相关性的 STEC 的 O 血清群：O26、O45、O91、O103、O111、O113、O121、O128、O145 和 O157。该方法采用 PCR 后结合 Luminex xMAP®技术，能够在单个多重反应中检测多种分析物，具有高通量能力。使用该检测方法，114 株 STEC 分离株得到了正确鉴定，在 46 种其他生物体中未出现假阳性。该检测方法在多个实验室中使用 Bio-Plex 200 和 MAGPIX 仪器上的 11 种不同 STEC 血清群的面板进行了性能测试。STEC 基于微球的悬浮阵列可在 96 孔板格式中进行，可在不到 4 小时内进行高通量筛选。此外，它具有可扩展性，如果需要，可以添加 O 血清群。
