On the planet Earth, the living of numerous species has strongly depended on microorganisms, one of which is bacteria. Among various bacteria, the role of pathogenic bacteria in terms of prevalent diseases, environmental disasters, and even animal life are undeniable. Indeed, in the modern era, diagnosis of the spread of these bacteria in different settings including the human body, food matrix, and environment can able us to control them. Despite the advancement of sensing platforms, there is a huge shortage in the development of sensitive and selective sensing approaches. Hence, fabrication efficient sensing-assay can play an important role. Amino acids (AAs) are part of biocompatible materials that have attracted attention as a promising class in various biosensors as sensing probes. This review focuses on the application of poly(AAs) such as cysteine and poly (Cysteine), lysine and poly (Lysine), glutamic acid and poly (Glutamic acid), glycine and poly (Glycine) and other types of poly(AAs) in terms of pathogenic bacteria detection. In addition, the role of different nanomaterials along with biological factors in increasing the sensitivity and selectivity of these types of assays will be reviewed. To elaborate, the identification of pathogenic bacteria will be investigated using different diagnostic platforms such as electrochemiluminescence (ECL), optical, electrochemical (EC), and photoelectrochemical (PEC) techniques, which will pay special attention to the role of AA and poly AA. Finally, the existing technological and research gaps will be examined and criticized in order to solve them in the future. It should be noted the prepared study is the first comprehensive review of bacterial assay detection by using poly(AAs) in the construction of biosensors.