This document describes a study that developed and validated a real-time PCR array to identify 87 antibiotic resistance genes from bacterial isolates and metagenomic samples. The array was used to profile resistance genes in Klebsiella pneumoniae isolates and human stool samples. A variety of resistance genes were detected, including SHV, KPC, ermB, mefA and tetA. The PCR array results were confirmed using pyrosequencing and shown to be effective for monitoring the spread of antibiotic resistance.