Sepsis presents a significant challenge in healthcare, with increasing incidence rates globally. This study addresses the pressing need for timely diagnosis and intervention by evaluating the efficacy of a rapid extraction and filtration method for  dentifying Gram-negative bacteria in blood samples. The research, conducted at Taipei Cathay General Hospital between July, 2023, and March 20, 2024, analyzed 191 positive blood bottle samples. The experimental group utilized Gram staining followed  by rapid extraction and filtration, while the control group underwent traditional subculture. Results showed a high identification agreement rate of 94% (177/188) between the experimental and control groups. Furthermore, drug susceptibility  testing exhibited a high level of consistency, with an Essential agreement (EA) and Category agreement (CA) of 98% (2694/2738) each. While the rapid method met acceptance criteria and demonstrated lower error rates, improvements are still warranted. This study underscores the potential of rapid diagnostic methods in improving sepsis management, with implications for enhancing patient care and healthcare efficiency. Future research should focus on broader clinical trials to further validate and optimize these diagnostic approaches.