Abstract <p>Biosensors are integrated devices intended for quantifying a biological or chemical reaction, that provide signals proportional to the analyte concentration in the reaction. The utilization of biosensors in probiotic determination has brought about a paradigm shift by offering a swift and precise approach to their identification. In recent years, probiotics that impart health benefits upon ingestion, have grown in popularity. However, identifying and quantifying them in a wide range of products can present difficulties. This review aims to summarize the latest research and challenges in the field of healthcare regarding probiotic determination. Biosensors enable rapid detection and quantification of biomarkers or metabolites linked to probiotics. The application of biosensors provides several benefits, including enhanced sensitivity and specificity, as well as the capability to quantify probiotic viability and activity directly. These methods can be applied to a wide range of matrices, encompassing food, beverages, and supplements, thereby providing significant data for the purposes of quality control and safety evaluation. The utilization of biosensors has facilitated progress in both probiotic research and industry by permitting the monitoring of probiotic content in various pharmaceutical and commercial products in a more dependable and efficient manner. The increasing need for probiotics will inevitably necessitate the development and application of biosensors for probiotic detection, which will be indispensable in guaranteeing their quality and effectiveness.</p>