Objective To analyze the tears of outpatients who had chronic conjunctivitis infected adenovirus (AdV) and (or) herpes simplex virus (HSV). Methods It was an experimental study. 81 patients with chronic conjunctivitis (adult 66 cases, children 15 cases ), 9 cases with acute viral conjunctivitis and 30 healthy control cases. To collect their tears, use polymerase chain reaction (PCR) to detect herpes simplex virus and adenovirus, and analyze the results of PCR, clinical symptoms and signs. Results In patients with chronic conjunctivitis, adenovirus-positive rate was 32. 1% (26/81) and herpes simplex virus-positive rate was 30. 9% ( 25/81 ). In children patients with chronic conjunctivitis, the adenovirus-positive rate was 33.3% (5/15) and herpes simplex virus-positive rate was 13. 3% (2/15). In adult group, the adenovirus-positive rate was 31.8% (21/66) and herpes simplex virus-positive rate was 34. 8% (23/66). In patients who underwent acute conjunctivitis, the adenovirus-positive rate was 61.5%(16/26). It has statistical differences ( χ2 = 16. 884, P ＜ 0. 01 ) with no acute conjunctivitis patients. The herpes simplex virus-positive rate was 42. 3% ( 11/26 ). It has statistical differences ( χ2 = 5. 351, P =0.021) with no acute conjunctivitis patients. The positive rate of virus detection was 0% (0/30) in the normal negative control group. The adenovirus-positive rate was 100. 0% (9/9) in the positive control grouppatients with acute viral conjunctivitis. 37 patients were adenovirus-positive and (or) herpes simplex viruspositive. The rate of redness with itching was 32. 4% (12/37) in virus positive patient. The rate of redness was 64.9% (24/37). The rate of itching was 56.8% (21/37). The rate of lower eyelid follicles was 73.0% (27/37) in virus positive patients. Conclusions The rate of adenovirus and herpes simplex virus positive expression is considerable in the patients with chronic conjunctivitis. The positive rate of patients with the history of virus infection is rather higher than no infection history.