Introduction: Ischemic stroke (IS) exhibits a high mortality rate, and its pathogenesis is intricately linked to genetic factors. This study was to examine the association between the precursor microRNA-218 (pre-miR-218) rs11134527 polymorphism and genetic susceptibility to IS in the Han Chinese population. Methods: The study included 320 IS patients and 290 healthy controls. Polymerase chain reaction (PCR) was utilized to identify the pre-miR-218 rs11134527 polymorphism. Quantitative real-time PCR was performed to measure the expression levels of miR-218-5p. Logistic regression analysis was conducted to identify independent risk factors for IS. Furthermore, a receiver operating characteristic (ROC) curve was constructed to evaluate the clinical diagnostic value of serum miR-218-5p levels in IS. Results: This study revealed that compared to the control group, the GG genotype (p < 0.001, OR = 2.277, 95% CI = 1.443–3.593) and AG genotype (p = 0.033, OR = 1.496, 95% CI = 1.032–2.168) at rs11134527 were significantly associated with an increased risk of IS. Individuals carrying the G allele (p < 0.001, OR = 1.526, 95% CI = 1.246–1.914) had an increased risk of developing IS. Furthermore, hypertension, hyperlipidemia, and the rs11134527 polymorphism were identified as independent risk factors for IS. The rs11134527 variation may influence lipid metabolism-related indices, potentially contributing to IS. Additionally, the GG genotype of rs11134527 correlated with a higher National Institutes of Health Stroke Scale (NIHSS) score, suggesting its potential for evaluating IS severity. Moreover, serum miR-218-5p levels were significantly elevated in IS patients. ROC curve analysis suggested that serum miR-218-5p could serve as a potential biomarker for diagnosing IS. Conclusion: The rs11134527 polymorphism in pre-miR-218 was strongly associated with genetic susceptibility to IS.

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