title: Low-Cost Charged-Coupled Device (CCD) Based Detectors for Shiga Toxins Activity Analysis.
abstract: To improve food safety there is a need to develop simple, low-cost sensitive devices for detection of food-borne pathogens and their toxins. We describe a simple, low-cost webcam-based detector which can be used for various optical detection modalities, including fluorescence, chemiluminescence, densitometry, and colorimetric assays. The portable battery-operated CCD-based detection system consists of four modules: (1) a webcam to measure and record light emission, (2) a sample plate to perform assays, (3) a light emitting diode (LED) for illumination, and (4) a portable computer to acquire and analyze images. To demonstrate the technology, we used a cell based assay for fluorescence detection of the activity of the food borne Shiga toxin type 2 (Stx2), differentiating between biologically active toxin and inactive toxin which is not a risk. The assay is based on Shiga toxin inhibition of cell protein synthesis measured through inhibition of the green fluorescent protein (GFP). In this assay, GFP emits light at 509 nm when excited with a blue LED equipped with a filter at 486 nm. The emitted light is then detected with a green filter at 535 nm. Toxin activity is measured through a reduction in the 509 nm emission. In this system the level of detection (LOD) for Stx2 was 0.1 pg/ml, similar to the LOD of commercial fluorometers. These results demonstrate the utility and potential of low cost detectors for toxin activity. This approach could be readily adapted to the detection of other food-borne toxins.
authors.affiliation: U.S. Department of Agriculture, Western Regional Research Center, Agricultural Research Service, 800 Buchanan St, Albany, CA, 94710, USA. reuven.rasooly@ars.usda.gov.
authors.forename: Reuven
authors.initials: R
authors.lastname: Rasooly
authors.affiliation: National Cancer Institute, National Institutes of Health, Rockville, MD, 208503, USA.
authors.forename: Ben
authors.initials: B
authors.lastname: Prickril
authors.affiliation: University of Maryland College Park (UMCP), College Park, MD, 20742, USA.
authors.forename: Hugh A
authors.initials: HA
authors.lastname: Bruck
authors.affiliation: National Cancer Institute, National Institutes of Health, Rockville, MD, 208503, USA.
authors.forename: Avraham
authors.initials: A
authors.lastname: Rasooly
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001681:Biological Assay; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods*; D018929:Cell Culture Techniques; D002460:Cell Line; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods; D057054:Molecular Imaging / Q000295:instrumentation / Q000379:methods; D055096:Optical Devices; D022603:Shiga Toxins / Q000494:pharmacology; D013223:Statistics as Topic / Q000379:methods; D014161:Transduction, Genetic
chemical_list: D022603:Shiga Toxins
keywords: CCD; Fluorescence; Fluorometer; GFP; LED; Microbial pathogens; Shiga toxin; Toxin activity; Webcam