Department of Cell Biology, Microbiology, and Molecular Biology, University of South Florida, Tampa, Florida 33620-5150, USA.
Appl Environ Microbiol. 2010 Dec;76(24):8044-52. doi: 10.1128/AEM.01990-10. Epub 2010 Oct 29.
Contamination of fresh produce with Escherichia coli O157:H7 and other pathogens commonly causes food-borne illness and disease outbreaks. Thus, screening for pathogens is warranted, but improved testing procedures are needed to allow reproducible same-day detection of low initial contamination levels on perishable foods, and methods for detecting numerous pathogens in a single test are desired. Experimental procedures were developed to enable rapid screening of spinach for E. coli O157:H7 by using multiplex-capable immunological assays that are analyzed using biosensors. Detection was achieved using an automated electrochemiluminescent (ECL) assay system and a fluorescence-based cytometric bead array. Using the ECL system, less than 0.1 CFU of E. coli O157:H7 per gram of spinach was detected after 5 h of enrichment, corresponding to 6.5 h of total assay time. Using the cytometric bead array, less than 0.1 CFU/g was detected after 7 h of enrichment, with a total time to detection of less than 10 h. These results illustrate that both biosensor assays are useful for rapid detection of E. coli O157:H7 on produce in time frames that are comparable to or better than those of other testing formats. Both methods may be useful for multiplexed pathogen detection in the food industry and other testing situations.
新鲜农产品中大肠杆菌 O157:H7 和其他病原体的污染通常会导致食源性疾病和疾病爆发。因此,有必要对病原体进行筛选,但需要改进测试程序,以允许在易腐食品上重现性地当天检测到低初始污染水平,并希望在单次测试中检测多种病原体的方法。已经开发了实验程序,以便通过使用生物传感器分析的可进行多重检测的免疫学检测来快速筛选菠菜中的大肠杆菌 O157:H7。使用自动化电化学发光(ECL)检测系统和基于荧光的细胞计数珠阵列进行检测。使用 ECL 系统,在富集 5 小时后,每克菠菜中可检测到少于 0.1 CFU 的大肠杆菌 O157:H7,相当于总检测时间 6.5 小时。使用细胞计数珠阵列,在富集 7 小时后可检测到少于 0.1 CFU/g,检测总时间不到 10 小时。这些结果表明,两种生物传感器检测都可用于在与其他检测方法可比或优于其他检测方法的时间范围内快速检测农产品上的大肠杆菌 O157:H7。这两种方法都可能有助于在食品工业和其他检测情况下进行多重病原体检测。