Antibiotic (AB) resistance is one of daunting challenges of our time, attributed to overuse of ABs and usage of AB-contaminated food resources. Due to their detrimental impact on human health, development of visual detection methods for multiplex sensing of ABs is a top priority. In present study, a colorimetric sensor array consisting of two types of gold nanoparticles (AuNPs) were designed for identification and determination of ABs. Design principle of the probe was based on aggregation of AuNPs in the presence of ABs at different buffer conditions. The utilization of machine learning algorithms in this design enables classification and quantification of ABs in various samples. The response profile of the array was analyzed using linear discriminant analysis algorithm for classification of ABs. This colorimetric sensor array is capable of accurate distinguishing between individual ABs and their combinations. Partial least squares regression was also applied for quantitation purposes. The obtained analytical figures of merit demonstrated the potential applicability of the developed sensor array in multiplex detection of ABs. The response profiles of the array were linearly correlated to the concentrations of ABs in a wide range of concentration with limit of detections of 0.05, 0.03, 0.04, 0.01, 0.06, 0.05 and 0.04 μg.mL–1 for azithromycin, amoxicillin, ciprofloxacin, clindamycin, cefixime, doxycycline and metronidazole respectively. The practical applicability of this method was further investigated by analysis of mixture samples of ABs and determination of ABs in river and underground water with successful verification.