title: Magnetic bead based immuno-detection of Listeria monocytogenes and Listeria ivanovii from infant formula and leafy green vegetables using the Bio-Plex suspension array system.
abstract: Listeriosis, a disease contracted via the consumption of foods contaminated with pathogenic Listeria species, can produce severe symptoms and high mortality in susceptible people and animals. The development of molecular methods and immuno-based techniques for detection of pathogenic Listeria in foods has been challenging due to the presence of assay inhibiting food components. In this study, we utilize a macrophage cell culture system for the isolation and enrichment of Listeria monocytogenes and Listeria ivanovii from infant formula and leafy green vegetables for subsequent identification using the Luminex xMAP technique. Macrophage monolayers were exposed to infant formula, lettuce and celery contaminated with L. monocytogenes or L. ivanovii. Magnetic microspheres conjugated to Listeria specific antibody were used to capture Listeria from infected macrophages and then analyzed using the Bio-Plex 200 analyzer. As few as 10 CFU/mL or g of L. monocytogenes was detected in all foods tested. The detection limit for L. ivanovii was 10 CFU/mL in infant formula and 100 CFU/g in leafy greens. Microsphere bound Listeria obtained from infected macrophage lysates could also be isolated on selective media for subsequent confirmatory identification. This method presumptively identifies L. monocytogenes and L. ivanovii from infant formula, lettuce and celery in less than 28 h with confirmatory identifications completed in less than 48 h.
authors.affiliation: U.S. Food and Drug Administration, Center for Food Safety and Applied Nutrition, 5100 Paint Branch Parkway, College Park, MD 20740, USA. Electronic address: james.day@fda.hhs.gov.
authors.forename: J B
authors.initials: JB
authors.lastname: Day
authors.affiliation: U.S. Army Medical Research Institute of Infectious Diseases, 1430 Veterans Drive Fort Detrick, MD 21702, USA.
authors.forename: U
authors.initials: U
authors.lastname: Basavanna
mesh_terms: D000818:Animals; D015373:Bacterial Typing Techniques / Q000295:instrumentation / Q000379:methods*; D002460:Cell Line; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D041943:Infant Formula / Q000737:chemistry*; D018545:Lactuca / Q000382:microbiology*; D008087:Listeria / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification* / Q000502:physiology; D008089:Listeria monocytogenes / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification* / Q000502:physiology; D008264:Macrophages / Q000737:chemistry / Q000382:microbiology; D060328:Magnetic Phenomena; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods*; D014675:Vegetables / Q000382:microbiology*
keywords: Bio-Plex; Detection; Foods; Listeria; Macrophage