Capture and identification of bacteria from fish muscle based on immunomagnetic beads and MALDI-TOF MS

Abstract

In the present study, E. coli was taken as a model bacterium, anti-E. coli functionalized magnetic beads were constructed and used to capture E. coli from aqueous extracts of fish sarcoplasmic protein (FSP) and fish muscle protein of sablefish. The excellency of the reproducibility of the present protocol was demonstrated by capturing E. coli from sablefish FSP extracts. The presence of 10 CFU/mL E. coli is still detectable. A microbial safety test on the surface of fish muscle was successfully performed. The bacterial identification accuracy from samples with different matrices was found to be excellent with RSD = 3%. High specific detection of target bacteria in complex biological samples was testified by spiking Staphylococcus aureus and Klebsiella pneumoniae in samples as interference. Ten biomarker ions were discovered for E. coli’s recognition. It is promising to apply the present protocol in bacterial analysis in muscle food samples to ensure their safety.

Keywords

• Escherichia coli
• Magnetic beads
• Sablefish
• matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)