Objective To analyze clinical characteristics of patients with re-positive of coronavirus disease 2019 (COVID-19), and provide guidance for epidemic prevention and control. Methods Clinical data of 30 patients who were re-admitted to the hospital due to re-positive of 2019-nCoV nucleic acid were collected, clinical data before and after re-positive was compared. Results Clinical symptoms were less when patients was re-positive of 2019-nCoV nucleic acid and re-admitted to the hospital, no fever occurred after re-positive,there were significant difference in fever, cough, throat-drying, sore throat, and main initial clinical symptoms before and after re-positive (all P<0.05). Most laboratory indicators of patients after re-positive and re-admission were normal, there were significant difference in lactate dehydrogenase (LDH), creatine kinase (CK), D-dimer, white blood cell (WBC), lymphocyte (LYM), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactic acid (LAC) before and after re-positive of 2019-nCoV nucleic acid (all P<0.05). There was no change in chest CT after re-positive in patients who had normal CT findings before re-positive, and the absorption was improved in patients who had pneumonia before re-positive. Length of re-hospitalization was shorter than that of first hospitalization ([6.93±3.84] days vs[17.73±7.22] days), difference was significant (P<0.05). Conclusion Condition of patients with re-positive of 2019-nCoV nucleic acid continues to improve, re-positive does not mean relapsing, there is no human- to-human transmission, and no serious public hazard has been caused, but the cause of re-positive of 2019-nCoV nucleic acid still needs further research.