Concentration and Detection of Salmonella in Mung Bean Sprout Spent Irrigation Water by Use of Tangential Flow Filtration Coupled with an Amperometric Flowthrough Enzyme-Linked Immunosorbent Assay

The development of a culture-free method for Salmonella screening of spent irrigation water derived from sprouting mung bean beds is described. The system used tangential flow filtration (TFF) to nonspecifically concentrate cells from large (2- to 10-liter) sample volumes. The retentate (100 ml) from the TFF was then flowed over an anti-Salmonella antibody-modified cellulose acetate membrane. The captured Salmonella was detected by reacting with a secondary anti-Salmonella and goat anti-rabbit biotin labeled antibody, followed by avidin-tagged glucose oxidase. The hydrogen peroxide generated from the enzymic oxidation of glucose was amperometrically detected at an underlying platinum electrode. It was found that 10 liters of Salmonella suspensions of 2 log CFU/ml could be concentrated to 4 log CFU/ml with 60% recovery regardless of the flow rate ( 112 to 511 ml/min) or transmembrane pressure (0 to 20 lb/in(2)) applied. The solids content of spent irrigation water negatively affected the filtration rate of TFF. This was most evident in spent irrigation water collected in the initial 24 h of the Sprouting period, where the Solids content was high (4,170 mg/liter) compared with samples collected at 96 h (560 mg/liter). Trials were performed using mung bean beds inoculated with different Salmonella levels (1.3 to 3.3 log CFU/g). By using the optimized TFF and flowthrough immunoassay it was possible to detect Salmonella in spent irrigation water at levels of 2.43 log CFU/ml within 4 h. The integrated concentration and detection system will provide a useful tool for sprout producers to perform in-house pathogen screening of spent irrigation water.