Escherichia coli Reference Center, Department of Veterinary and Biomedical Sciences, Pennsylvania State University, University Park, PA 16802, USA.
J Microbiol Methods. 2013 Jun;93(3):273-6. doi: 10.1016/j.mimet.2013.03.024. Epub 2013 Apr 6.
An antibody microarray was developed to detect the "top six" non-O157 serogroups, O26, O45, O103, O111, O121, and O145 of Shiga toxin-producing Escherichia coli (STEC), that have been declared as adulterant in meat by the Food Safety and Inspection Service of the United States Department of Agriculture. The sensitivity of the array was 10(5)CFU and the limit of detection of each serogroup in artificially inoculated ground beef was 1-10 CFU following 12h of enrichment. Optimal concentrations of antibodies for printing and labeling and bacterial dilutions for binding to the antibodies were assessed. The array utilized a minimal amount of antibodies and other reagents and may be utilized for screening of multiple target O groups of STEC in parallel, directly from enriched samples in less than 3h. Furthermore, the antibody array provides the flexibility to include other O serogroups of E. coli and may be adopted for high throughput screening. The method is potentially applicable to detect the pathogenic STEC O groups of E. coli in meat and other food, thus improving food safety and public health.
开发了一种抗体微阵列来检测“六大”非 O157 血清群,即美国农业部食品安全检验局宣布的肉类掺杂物的产志贺毒素大肠杆菌(STEC)的 O26、O45、O103、O111、O121 和 O145。该阵列的灵敏度为 10(5)CFU,在 12 小时的富集后,人工接种的绞碎牛肉中每个血清群的检测限为 1-10 CFU。评估了用于打印和标记的最佳抗体浓度以及与抗体结合的细菌稀释度。该阵列使用了最少的抗体和其他试剂,并且可以直接从浓缩样品中在不到 3 小时内平行筛选多个 STEC 目标 O 群。此外,抗体阵列提供了包含其他大肠杆菌 O 血清群的灵活性,并且可以用于高通量筛选。该方法有可能适用于检测肉类和其他食品中的致病性 STEC O 群大肠杆菌,从而提高食品安全和公共健康。