A disposable electrochemical immunosensor arrays using 4-channel screen-printed carbon electrode for simultaneous detection of Escherichia coli O157:H7 and Enterobacter sakazakii

An electrochemical immunosensor for Escherichia coli O157:H7 (E. coli O157:H7) and Enterobacter sakazakii (E. sakazakii) detection using carbon screen-printed low-density arrays is reported. The sensors were fabricated based on screen-printed carbon arrays containing four carbon working electrode, an integrated carbon counter electrodes and an integrated Ag/AgCl reference electrode. Multi-walled carbon nanotubes (MWCNTs)/sodium alginate (SA)/carboxymethyl chitosan (CMC) composite films were coated on all the working electrodes to enhance the sensitization of the electrode. Horseradish peroxidases (HRP) labeled antibodies of two bacteria were immobilize on different working electrode of the same screen-printed electrode respectively. The immobilization of MWCNTs, HRP labeled antibodies onto the screen-printed carbon electrodes was examined using atom force microscopy (AFM) and cyclic voltammetry (CV). The analytical performance of proposed immunosensor arrays toward E. sakazakii and E. coli O157:H7 was investigated by AFM and CV. Under optimal conditions, the linear range of E. sakazakii and E. coli O157:H7 were from 104 to 1010 cfu/ml, with a detection limit of 4.57 × 103 cfu/ml (S/N = 3) and 3.27 × 103 cfu/ml (S/N = 3), respectively. The specificity, reproducibility, stability and accuracy of the proposed immunosensor arrays were also evaluated. Two antibodies modified work electrodes were tested and compared in terms of sensitivity and ability to recognize different pathogenic biological species.