To assess the efficacy of targeted therapy based on drug sensitivity testing (DST) results in patients with acute pulpitis (AP).A total of 80 cases of AP were included retrospectively and divided into two groups: control (Ctrl) group (conventional drug palliative therapy, n=40) and experimental (Exp) group (DST + non-resistant drugs, n=40). The clinical data and laboratory examination data of patients, including bacterial culture data, drug sensitivity test results, Ca and P contents in dental pulp, visual analogue scale (AVS), treatment satisfaction, and dental pulp incidence, were collected and analyzed.Prevotella melaninogenica, Fusobacterium nucleatum, and Porphyromonas gingivalis exhibited higher resistance rates (RS) to penicillin and amoxicillin but no resistance to imipenem and metronidazole. The content of Ca and P in the dental pulp of the Exp group patients was significantly higher than that of the Ctrl group (P=0.006). The total response rate (95% vs. 77.5%, P=0.018) and overall patient satisfaction (92.5% vs. 80%, P=0.021) were also significantly higher in the Exp group than in the Ctrl group. Additionally, when the follow-up duration exceeded 1 year, the incidence of dental pulp reactions in the Exp group was significantly lower than that of the Ctrl group (P=0.026).These findings suggest that Gram-negative anaerobes are the predominant oral pathogens in patients with AP. Imipenem and metronidazole demonstrate the most effective anti-infective properties against these anaerobes. Utilizing DST to select non-resistant drugs for treatment prior to therapy effectively enhances clinical outcomes for patients with AP.