Background: The duration for COVID-19 patients to achieve a negative nucleic acid test result is influenced by various factors. This study aimed to explore the influences of clinical symptoms of COVID-19 on the time required for nucleic acid test results to become negative. Methods: The retrospective study enrolled a total of 486 participants infected with COVID-19 who were aged ≥18 years and hospitalized at Shanghai Lingang Make-shift Hospital between April 10, 2022, and May 20, 2022. The clinical data included patient demographics, clinical symptoms, and the time of nucleic acid conversion to negative, all assessed using a cross-sectional study method. Results: This study included COVID-19 patients with a median age of 38, comprising 66.0% males and 34.0% females. Multiple linear regression analyses demonstrated a significant positive correlation between the time of nucleic acid conversion to negative and the clinical symptoms of throat pain among patients with COVID-19 ( β=0.845, 95% CI: 0.123–1.567, P≤0.05). Logistic regression analysis showed that throat pain had a significant negative effect on the 7-day negative nucleic acid conversion rate (OR=0.586, 95%CI:0.390-0.880, P≤0.01), as did diarrhea（OR = 0.541, 95%CI=0.295-0.989,P≤0.05）. Conclusion: The varying clinical symptoms influence the time to negative nucleic acid test results in COVID-19 patients, indicating that these symptoms should receive special attention in clinical treatment to optimize therapy and shorten recovery time.