Acute kidney injury (AKI) is a common and often fatal condition with a high risk of mortality. Standard metrics like serum creatinine and blood urea nitrogen levels are insensitive and delay diagnosis, affecting timely patient management and preclinical toxicity evaluation. The development of sensitive, specific, and reliable biomarkers for early diagnosis and prognosis of AKI, along with high-throughput technologies for rapid multiplexed detection, is crucial for improving AKI management. Key biomarkers discussed include N-acetyl-β-glucosaminidase (NAG), β2-microglobulin (β2M), α1-microglobulin (α1M), retinol binding protein (RBP), cystatin-C, microalbumin, kidney injury molecule-1 (KIM-1), clusterin, neutrophil gelatinase-associated lipocalin (NGAL), interleukin-18 (IL-18), cysteine-rich protein (Cyr61), osteopontin, fatty acid-binding protein (FABP), and sodium/hydrogen exchanger isoform (NHE3). These biomarkers have shown promise in early detection and monitoring of AKI, but further research is needed to validate their utility in clinical settings. Advances in microfluidics and nanotechnology offer potential for rapid, sensitive detection methods, which could transform AKI diagnosis and treatment.Acute kidney injury (AKI) is a common and often fatal condition with a high risk of mortality. Standard metrics like serum creatinine and blood urea nitrogen levels are insensitive and delay diagnosis, affecting timely patient management and preclinical toxicity evaluation. The development of sensitive, specific, and reliable biomarkers for early diagnosis and prognosis of AKI, along with high-throughput technologies for rapid multiplexed detection, is crucial for improving AKI management. Key biomarkers discussed include N-acetyl-β-glucosaminidase (NAG), β2-microglobulin (β2M), α1-microglobulin (α1M), retinol binding protein (RBP), cystatin-C, microalbumin, kidney injury molecule-1 (KIM-1), clusterin, neutrophil gelatinase-associated lipocalin (NGAL), interleukin-18 (IL-18), cysteine-rich protein (Cyr61), osteopontin, fatty acid-binding protein (FABP), and sodium/hydrogen exchanger isoform (NHE3). These biomarkers have shown promise in early detection and monitoring of AKI, but further research is needed to validate their utility in clinical settings. Advances in microfluidics and nanotechnology offer potential for rapid, sensitive detection methods, which could transform AKI diagnosis and treatment.