4.6 Article

Rapid Sample Processing for Detection of Food-Borne Pathogens via Cross-Flow Microfiltration

Journal

APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Volume 79, Issue 22, Pages 7048-7054

Publisher

AMER SOC MICROBIOLOGY
DOI: 10.1128/AEM.02587-13

Keywords

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Funding

  1. U.S. Department of Agriculture (USDA) [935-42000-049-00D]
  2. Purdue University Agricultural Research Programs
  3. Department of Agricultural and Biological Engineering at Purdue University

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This paper reports an approach to enable rapid concentration and recovery of bacterial cells from aqueous chicken homogenates as a preanalytical step of detection. This approach includes biochemical pretreatment and prefiltration of food samples and development of an automated cell concentration instrument based on cross-flow microfiltration. A polysulfone hollow-fiber membrane module having a nominal pore size of 0.2 mu m constitutes the core of the cell concentration instrument. The aqueous chicken homogenate samples were circulated within the cross-flow system achieving 500- to 1,000-fold concentration of inoculated Salmonella enterica serovar Enteritidis and naturally occurring microbiota with 70% recovery of viable cells as determined by plate counting and quantitative PCR (qPCR) within 35 to 45 min. These steps enabled 10 CFU/ml microorganisms in chicken homogenates or 10(2) CFU/g chicken to be quantified. Cleaning and sterilizing the instrument and membrane module by stepwise hydraulic and chemical cleaning (sodium hydroxide and ethanol) enabled reuse of the membrane 15 times before replacement. This approach begins to address the critical need for the food industry for detecting food pathogens within 6 h or less.

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