Background: The early diagnosis of non-ST-segment elevation myocardial infarction (NSTEMI) in patients with chronic kidney disease (CKD) remains a challenge. Methods: The study consecutively enrolled patients who had suffered from chest pain within 3 h whose electrocardiogram had no elevation in the ST segment. CKD was defined as an estimated glomerular filtration rate (eGFR) <60 mL/min/1.73 m<sup>2</sup>, and the diagnostic criteria for NSTEMI were defined according to the recommended guideline. Circulating microRNA-1 was collected and determined by quantitative real-time reverse transcription polymerase chain reaction. Results: A total of 456 patients with suspected NSTEMI were included. There were 115 patients in the CKD group, including 67 with NSTEMI, 20 with stable angina, 7 with unstable angina, 18 with heart failure, and 3 with other disorders. Compared with the NSTEMI group, the non-NSTEMI group just had significant differences in microRNA-1 and high-sensitivity cardiac troponin I (hs-cTnI) (both p < 0.05). The relative expression of microRNA-1 was significantly increased in the NSTEMI group as compared with that in the other disease groups (all p < 0.05). A receiver operating characteristic (ROC) curve analysis suggested that microRNA-1 and hs-cTnI had advantages in the early diagnosis of NSTEMI with CKD (AUC [area under the ROC curve] 0.879 and 0.812, respectively, both p < 0.05). Compared with that in the non-CKD group, the accuracy of microRNA-1 was almost as good in the CKD group (84.3 vs. 89.4%, p > 0.05). However, the diagnostic accuracy of hs-cTnI was significantly decreased (79.1 vs. 91.5%, p < 0.05), as was its specificity (75.0 vs. 95.5%, p < 0.05). There was no significant difference in the correlation between microRNA-1 and eGFR (p > 0.05), but a statistically significantly negative correlation between hs-cTnI and eGFR (p < 0.05). Conclusion: Circulating microRNA-1 is capable of early diagnosis of NSTEMI in patients with CKD suffering from chest pain.