To explore the flora characteristics and differences of esophageal tissues between elderly esophageal squamous cell carcinoma (ESCC) patients and young and middle-aged ESCC patients, so as to assist in studying the potential biomarkers of elderly ESCC patients. In this study, a retrospective study was adopted. 72 ESCC patients diagnosed in Taihe Hospital, Shiyan City, Hubei Province from July 2018 to July 2019 were selected, including 49 patients in the elderly group (≥ 60 years old, 40 males and 9 females), 23 patients in the young and middle-aged group (0.05). There was no significant difference in β-diversity between the elderly group and the young and middle-aged group (PC1=19.14%, PC2=6.95%, =0.67, =0.42). At the phyla level, the top 5 phyla in abundance were as follows: and in the young and middle-aged group, while the top 5 phyla in abundance were as follows: and in the elderly group; the significant difference between the two groups was (=0.596, 0.05). At the genus level, the top 5 genera in the young and middle-aged group in abundance were as follows: and In the elderly group, and were the top 5 in abundance, and there were significant difference in between the two groups (=0.938, 0.05). PICRUSt function prediction showed that the abundance of and in the elderly group were lower than those in the young and middle-aged group (all =0.734, 0.05). There is no significant difference in α-diversity and β-diversity between elderly ESCC patients and young and middle-aged patients, but the abundance of flora increased.