Same-Day Detection of Escherichia coli O157:H7 from Spinach by Using Electrochemiluminescent and Cytometric Bead Array Biosensors

• Published in: Applied and Environmental Microbiology 2010-12, Vol.76 (24), p.8044-8052
• Main Authors: Leach, Kelly M, Stroot, Joyce M, Lim, Daniel V
• Format: Article
• Language: English
• Subjects: Bacteriological Techniques - methods; Biological and medical sciences; Biosensing Techniques - methods; Biosensors; Biotechnology; E coli; Escherichia coli; Escherichia coli O157 - isolation & purification; Flow cytometry; Flow Cytometry - methods; Flowers & plants; Fundamental and applied biological sciences. Psychology; Immunoassay - methods; Luminescence; Luminescent Measurements - methods; Methods; Methods. Procedures. Technologies; Microbiology; Pathogens; Research methodology; Sensitivity and Specificity; Spinacia oleracea; Spinacia oleracea - microbiology; Test methods; Time Factors; Various methods and equipments
• ISSN: 0099-2240; 1098-5336; 1098-6596
• DOI: 10.1128/AEM.01990-10

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.