Objective To investigate the application value of detection of serum D-dimer and procalcitonin levels in sepsis. Methods 78 patients with sepsis admitted to our hospital from May 2019 to May 2022 were selected as the observation group, and 78 patients who underwent physical examination in our hospital during the same period of time were collected as the control group. Serum D-dimer and procalcitonin levels of the two groups were detected, and serum D-dimer and procalcitonin levels of the observation group before and after treatment were detected. The observation group was divided into 10 cases of septic shock (observation group A), 15 cases of severe sepsis (observation group B) and 53 cases of sepsis (observation group C). The differences of serum D-dimer and procalcitonin levels among the three groups were compared. Results The serum D-dimer and procalcitonin were detected in the observation group, and the difference was statistically significant compared with the control group (P<0.05). After treatment, two indexes (serum D-dimer and procalcitonin) in the observation group showed significant changes, and the difference was statistically significant compared with before treatment (P<0.05). The observation group was divided into three groups, and the results showed that the data of observation group C was the lowest, followed by observation group B, and the highest was observation group A, and the difference between groups was statistically significant (P<0.05). Conclusion The detection of serum D-dimer and procalcitonin levels is of high application value in sepsis.