Lateral Flow Immunosensing of Salmonella Typhimurium Cells in Milk: Comparing Three Sequences of Interactions

To ensure the safety of foodstuffs, widespread non-laboratory monitoring for pathogenic contaminants is in demand. A suitable technique for this purpose is lateral flow immunoassay (LFIA) which combines simplicity, rapidity, and productivity with specific immune detection. This study considered three developed formats of LFIA for <i>Salmonella</i> Typhimurium, a priority pathogenic contaminant of milk. Common sandwich LFIA with all immunoreagents pre-applied to the test strip (format A) was compared with incubation of the sample and (gold nanoparticle-antibody) conjugate, preceding the lateral flow processes (format B), and sequential passages of the sample and the conjugate along the test strip (format C). Under the chosen conditions, the detection limits and the assay times were 3 × 10⁴, 1 × 10⁵, and 3 × 10⁵ cells/mL, 10, 15, and 20 min for formats A, B, and C, respectively. The selected format A of LFIA was successfully applied to test milk samples. The sample's dilution to a fat content of 1.0% causes pathogen detection, with 70-110% revealing and 1.5-8.5% accuracy. The obtained results demonstrate that the developed LFIA allows the detection of lower concentrations of <i>Salmonella</i> cells and, in this way, accelerates decision-making in food safety control.