Automated Immunomagnetic Separation and Microarray Detection of E. coli O157:H7 from Poultry Carcass Rinse

Journal Article · Sat Sep 01 04:00:00 EDT 2001 · International Journal of Food Microbiology

DOI:https://doi.org/10.1016/S0168-1605(01)00536-0· OSTI ID:15005639

Chandler, Darrell P ^[1]; Brown, Jeremy D ^[2]; Call, Douglas R ^[1]; Wunschel, Sharon C ^[1]; Grate, Jay W ^[1]; Holman, David A ^[2]; Olson, Lydia G ^[2]; Stottlemyer, Mark S ^[3]; Bruckner-Lea, Cindy J ^[1]

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1. BATTELLE (PACIFIC NW LAB)
2. ASSOC WESTERN UNIVERSITY
3. Associated Western Universities

We describe the development and application of a novel electromagnetic flow cell and fluidics system for automated immunomagnetic separation of E. coli directly from unprocessed poultry carcass rinse, and the biochemical coupling of automated sample preparation with nucleic acid microarrays without cell growth. Highly porous nickel foam was used as a magnetic flux conductor. Up to 32% recovery efficiency of 'total' E. coli was achieved within the automated system with 6 sec contact times and 15 minute protocol (from sample injection through elution), statistically similar to cell recovery efficiencies in > 1 hour 'batch' captures. The electromagnet flow cell allowed complete recovery of 2.8 mm particles directly from unprocessed poultry carcass rinse whereas the batch system did not. O157:H7 cells were reproducibly isolated directly from unprocessed poultry rinse with 39% recovery efficiency at 103 cells ml-1 inoculum. Direct plating of washed beads showed positive recovery of O 157:H7 directly from carcass rinse at an inoculum of 10 cells ml-1. Recovered beads were used for direct PCR amplification and microarray detection, with a process-level detection limit (automated cell concentration through microarray detection) of < 103 cells ml-1 carcass rinse. The fluidic system and analytical approach described here are generally applicable to most microbial detection problems and applications.

• Research Organization: Pacific Northwest National Lab., Richland, WA (US)
• Sponsoring Organization: US Department of Energy (US)
• DOE Contract Number: AC06-76RL01830
• OSTI ID: 15005639
• Report Number(s): PNWD-SA-5123
• Journal Information: International Journal of Food Microbiology, Journal Name: International Journal of Food Microbiology Journal Issue: 1-2 Vol. 70
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
AMPLIFICATION
DETECTION
EFFICIENCY
ELECTROMAGNETS
FOWL
MAGNETIC FLUX
NICKEL
NUCLEIC ACIDS
PLATING
SAMPLE PREPARATION
SENSITIVITY