Biotin exposure–based immunomagnetic separation coupled with sodium dodecyl sulfate, propidium monoazide, and multiplex real-time PCR for rapid detection of viable Salmonella Typhimurium, Staphylococcus aureus, and Listeria monocytogenes in milk

In this study, we established a rapid and sensitive method for the detection of viable Salmonella Typhimurium, Staphylococcus aureus, Listeria monocytogenes in milk using biotin-exposure-based immunomagnetic separation (IMS) combined with sodium dodecyl sulfate (SDS), propidium monoazide (PMA), multiplex real-time PCR (mRT-PCR). We used IMS to lessen assay time isolation target bacteria. then optimized coupling conditions capture process. The immunoreaction incubation times 5 μg mAb coupled 500 streptavidin-functionalized magnetic beads streptavidin-biotin system were 90 30 min, respectively. Treatment SDS-PMA before mRT-PCR amplification eliminated false-positive outcomes from dead bacteria identified good sensitivity specificity. limit SDS-PMA-mRT-PCR Staph. L. spiked matrix samples was 10 cfu/mL remained significant even appearance 106 nontarget entire process able identify within 9 h. combination biotin-exposure-mediated has potential value foodborne pathogens.