Listeriosis through contaminated food is one of the leading causes of premature deaths in pregnant women and new born babies. Here, the authors have developed a magnetosomes-based biosensor for the rapid, sensitive, specific and costeffective detection of Listeria monocytogenes from food sample. Magnetosomes were extracted from Magnetospirillum sp. RJS1 and then directly bound to anti-Listeriolysin antibody (0.25-1 μg/ml), confirmed in spectroscopy. Listeriolysin (LLO) protein (0.01-7 μg/ml) was optimised in enzyme-linked immunosorbent assay. Magnetosomes was conjugated with LLO antibody (0.25 μg/ml) in optimum concentration to detect LLO protein (0.01 μg/ml). Magnetosomes-LLO antibody complex was 25% cost effective. The magnetosomes-LLO antibody complex was directly stabilised on screen printed electrode using external magnet. The significant increase in resistance (RCT value) on the electrode surface with increase in concentration of LLO protein was confirmed in impedance spectroscopy. The L. monocytogenes contaminated milk and water sample were processed and extracted LLO protein was detected in the biosensor. The specificity of the biosensor was confirmed in cross-reactivity assay with other food pathogens. The detection limit of 101 Cfu/ml in both water and milk sample manifests the sensitive nature of the biosensor. The capture efficiency and field emission scanning electron microscopy confirmed positive interaction of Listeria cells with magnetosomes-antibody complex. © 2020 Institution of Engineering and Technology. All rights reserved.