Rapid Quantification of Escherichia coli O157:H7 and Salmonella Typhimurium in Lettuce Using Immunomagnetic Separation and a Microfluidic System

Biol Pharm Bull. 2024;47(11):1931-1936. doi: 10.1248/bpb.b24-00352.

Abstract

Escherichia coli O157:H7 and Salmonella spp. are common foodborne pathogens. Simple, rapid and accurate methods to detect and enumerate these pathogens are required to prevent outbreaks of foodborne illness. Here, a microfluidic system with on-chip staining and semi-automated counting functionality was combined with the use of immunomagnetic separation to collect E. coli O157:H7 and Salmonella Typhimurium without an enrichment step. The recovery of bacteria from lettuce at different spiking ratios of the two species was 61-83%. In the range 3.5 × 102 to 3.3 × 105 cells/g of lettuce, there was a good linear relationship (R2 = 0.9997-0.9998) between the average counts from the microfluidic quantification method and conventional immunofluorescence microscopy. Detection took <3 h. The limit of detection of the microfluidic device was 4 × 101 cells/g, comparable to that of plate culture and real-time PCR methods. Our microfluidic approach has potential for rapid on-site detection of multiple pathogenic bacteria in foods.

Keywords: Escherichia coli; Salmonella Typhimurium; foodborne pathogen; immunomagnetic separation; microfluidic device; rapid quantification.

MeSH terms

  • Escherichia coli O157* / isolation & purification
  • Food Microbiology* / methods
  • Immunomagnetic Separation* / instrumentation
  • Immunomagnetic Separation* / methods
  • Lab-On-A-Chip Devices
  • Lactuca* / microbiology
  • Microfluidic Analytical Techniques* / instrumentation
  • Microfluidic Analytical Techniques* / methods
  • Salmonella typhimurium* / isolation & purification