Abstract: OBJECTIV To study the value of nucleic acid detection in diagnosis of patients with imported Zika virus disease in China and explore the applicability of the nucleic acid in detection of Zika virus in clinical samples. METHODS The blood, sputum, and urine specimens were collected from the patients with imported Zika virus disease who were treated in hospitals from Feb 2016 to Oct 2016. The clinical symptoms, signs, and results of clinical laboratory tests were recorded and observed. RESULTS The positive rate of initial detection of Zika virus nucleic acid was 61.5% in blood, 76.9% in sputum, 76.9% in urine. The total positive rate of detection of nucleic acid was 61.5% in blood, 84.6% in sputum, 100% in urine. The average duration that the nucleic acid maintained positive was 2.62 days in blood, 5.69 days in sputum, 11.62 days in urine; there was no case whose blood maintained positive for more than 1 week and no case whose sputum maintained positive for 10 days, however, there were 11 cases whose urine maintained positive for more than 1 week, including 6 cases whose urine maintained positive for more than 10 days. With the development of disease course, the detection rate was reduced in the samples. The positive rate of nucleic acid was 25% in blood on Day 3 after onset, 100% in sputum, 100% in urine; the positive rate of nucleic acid was 7.7% in blood on Day 7 after onset, 46.2% in sputum, 84.6% in urine. Few urine specimens showed hysteresis phenomenon to be tested positive. CONCLUSION The detection of Zika virus nucleic acid in urine has the advantages such as the high detection rate and long duration and should be used as the major method for clinical diagnosis of Zika virus disease, however, it is still necessary to collect the blood, sputum, and urine samples in early stage of the onset for simultaneous detection of the Zika virus nucleic acid.