Immunoliposome-based immunomagnetic concentration and separation assay for rapid detection of Cronobacter sakazakii

Abstract

This study aimed to develop an immunoliposome-based immunomagnetic concentration and separation assay for the rapid detection of Cronobacter sakazakii (C. sakazakii), an acute opportunistic foodborne pathogenic bacterium, in both pure culture and infant formula. To develop the assay, magnetic nanoparticles (diameter 30nm) were coated with immunoglobulin G (IgG), specifically anti-C. sakazakii IgG, and applied for the sensitive and efficient detection of C. sakazakii using immunoliposomes. The binding efficiency of anti-C. sakazakii IgG to the magnetic nanoparticles was 86.23±0.59%. The assay developed in this study detected as few as 3.3×103CFUmL−1 of C. sakazakii in pure culture within 2h 30min; in comparison, an indirect non-competitive enzyme-linked immunosorbent assay was able to detect 6.2×105CFUmL−1 of C. sakazakii in pure culture after 17h. The developed assay did not show any cross-reactivity with other Cronobacter spp. or pathogens belonging to other genera. In addition, the method was able to detect 103CFUmL−1 of C. sakazakii in infant formula without any pre-incubation. These results confirm that the immunoliposome-based immunomagnetic concentration and separation assay may facilitate highly sensitive, efficient, and rapid detection of C. sakazakii.