Chemiluminescence imaging immunoassay for multiple aminoglycoside antibiotics in cow milk

Abstract

SummaryA simple and rapid multiplexed direct competitive chemiluminescent (CL) imaging immunoassay had been developed for the simultaneous detection of kanamycin (KAN) and streptomycin (STR), which were widely used against bacterial infections in animals. To achieve the multiplexed detection of the two targets, a microarray format based on nitrocellulose membranes (NCMs) has been designed. Then, the peroxidase activity of the captured HRP‐labelled immunocomplex in each channel was measured by an enhanced luminol‐based chemiluminescent solution using a cooled low‐light CCD with high resolution. For qualitative analysis, the limit of detection (LOD) was 0.5 ng mL−1 for KAN and 3.13 ng mL−1 for STR by the naked eye. Through the CL data collected, KAN and STR could be detected quantitatively at the LOD of 0.03 and 0.33 ng mL−1, respectively. This method can be utilised as a screening test to evaluate the presence of antibiotics in milk samples.