Rapid Flow Cytometric Detection of Single Viable Salmonella Cells in Milk Powder

Abstract

Salmonella, a highly virulent food-borne pathogen transmitted through food, can cause severe infectious diseases in a large number of people through a single outbreak, due to its low infective doses. In this study, a flow cytometry (FCM)-based method was developed for the rapid detection of single viable Salmonella cells with dual staining of fluorescein isothiocyanate (FITC)-labeled anti-Salmonella antibody and propidium iodide (PI) dyes. The FCM-based method includes 6 h of pre-enrichment, 40 min of target cell isolation, and 20 min of dual staining and FCM analysis. The developed method demonstrated high specificity for the detection of 23 Salmonella strains and 22 food-borne pathogenic non-Salmonella strains. Furthermore, the analyses of 30 samples of milk powder artificially contaminated with single Salmonella cells, 123 samples of retail milk powder, and 6 samples of Salmonella-positive milk powder were performed by the FCM-based as well as traditional plate-based methods for testing the efficiency of the methods. The two methods yielded similar results for the detection of pathogens in all milk powder samples. In conclusion, the developed FCM-based method was found to be efficient in detecting single viable Salmonella cells in milk powder within 7 h. The proposed dual-color FITC assay combined with pre-enrichment offers a great potential for the rapid and sensitive detection of other pathogens in dairy products.