The present study elucidated the pathogenesis of allergic symptoms (AS) related to contact lens (cL) wear by assaying cL care solutions in lens storage cases and tears from subjects with AS using molecular biology techniques. A total of 15 cL storage cases were collected from subjects with AS (n=9) and healthy, asymptomatic control cL wearers (n=6). Bacterial populations in cL care solutions and tears were assayed by culture and 16S rdNA sequencing. Histamine levels in tears were measured by high-performance liquid chromatography. Western blot analysis was performed to identify the bacteria recognized by tear IgE from subjects with AS. No significant differences were found in the culture results between the subjects with AS and asymptomatic subjects. Histamine was detected in 2 subjects with AS. Meta-16S rDNA sequencing identified a cluster of 4 subjects with AS that were distinguished from others by principal coordinate analysis. detailed population analysis revealed that the abundance of Gram-positive bacteria in the microbiomes of cL care solutions used by the subjects with AS were higher than those of asymptomatic subjects (42.24±9.47 vs. 16.85±22.76% abundance). Among these, Streptococcus was the dominant genus (12.1-18.3% abundance). Tear microbiome analysis revealed that the abundance of Streptococcus in the subjects with AS was significantly higher than that in other subjects (19.02±5.50 vs. 3.08±3.35%, P<0.01). Western blot analysis demonstrated that the tear IgE in all subjects with AS reacted with Streptococcus (100%), but not with Staphylococcus. On the whole, these results provide novel insight into the pathogenesis of AS and identify Streptococcus as an important factor in AS associated with cL wear.