Electrochemical coupled immunosensing platform based on graphene oxide/gold nanocomposite for sensitive detection of Cronobacter sakazakii in powdered infant formula

Abstract

A sensitive electrochemical immunosensing platform for the detection of Cronobacter sakazakii was developed using a graphene oxide/gold (GO/Au) composite. Transmission electron microscopy showed that the Au nanoparticles, with an average size of < 30 nm, were well dispersed on the GO surface. For the detection of C. sakazakii, a polyclonal anti-C. sakazakii antibody (IgG) was covalently immobilized to the Au nanoparticles on the surface of the GO/Au composite coated glassy carbon electrode (GCE). The electrochemical sensing performance of immunofunctionalized GCE was characterized by cyclic voltammetry and differential pulse voltammetry. Under optimized conditions, in pure culture there was a linear relationship between electrical signal and C. sakazakii levels over the range 2.0 × 10²-2.0 × 10⁷ cfu/mL (R² = 0.999), with a detection limit of 2.0 × 10¹ cfu/mL. The total analytical time was 15 min per sample. The C. sakazakii electrochemical immunosensing assay was able to successfully detect 2.0 × 10¹ cfu/mL of C. sakazakii in artificially contaminated powdered infant formula without any enrichment or pre-enrichment steps. Furthermore, the recovery rates of the C. sakazakii electrochemical immunosensing assay following spiking of powdered infant formula with different concentrations of C. sakazakii (cfu/mL) were 82.58% at 2.0 × 10¹ cfu/mL, 84.86% at 2.0 × 10² cfu/mL, and 95.40% at 2.0 × 10³ cfu/mL. The C. sakazakii electrochemical immunosensing assay had good selectivity, reproducibility, and reactivity compared with other Cronobacter spp. and/or pathogens belonging to other genera, indicating its significant potential in the clinical diagnosis of C. sakazakii.

Keywords: Cronobacter sakazakii; Electrochemical immunosensing; Graphene oxide–gold composite; Pathogen detection; Powdered infant formula.