The rapid and sensitive detection of pathogenic bacteria is becoming increasingly important for the timely prevention of contamination and treatment of infections. Biosensors based on nucleic acid aptamers integrat-ed with optical, electrochemical, and mass-sensitive analytical techniques have garnered intense interest for their versatility, cost-efficiency, and ability to exhibit high affinity and specificity in binding bacterial bi-omarkers, toxins, and whole cells. This review emphasizes the development of aptamers, their structural op-timization, and their use in bacterial sensing applications. Moreover, we explore barriers and develop per-spectives in applying aptamer-based bacterial detection.