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An immunochromatographic test was developed for the simultaneous detection of several compounds in a complex sample matrix. The system was designed in a ‘traffic light’ format comprising three lines of different colors on a test strip, thereby providing an easy tool with which to identify an analyte of interest based on the visible color of the line formed (qualitative analysis), and to determine the amount of the analytes present based on the fluorescence intensity of the lines (quantitative analysis). For the development of the multicolor immunochromatographic test, we used antibodies against antibiotics of three different classes as selective binders. Each antibody was labeled with water-soluble quantum dots with emission maximum at either 525, 585, or 625nm. The test system exhibited high sensitivity, with limits of detection for ofloxacin, chloramphenicol, and streptomycin of 0.3, 0.12, and 0.2ngmL−1, respectively. These values are 80–200 times lower than those achievable with ELISA using the same antibodies. Using the ‘traffic light’ assay, these antibiotics could be detected in milk samples within 10min without any sample preparation. The ‘traffic light’ assay also demonstrated a high degree of analyte detection when testing spiked milk samples (92–101%) and accuracy (quantitation error <8% of the mean).
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