Background: Hemodialysis (HD) is a public health issue, and HD induced infection is a major risk factor for mortality. Toll like receptor 4 (TLR4) plays a key role in the pathogenesis of infection. However, polymorphisms of TLR4 contribute to infection are still unknown. Aim: This study aimed to examine the correlation between polymorphisms of TLR4 and mortality among HD patients. Methods: This retrospective cohort study included 585 participants from Tri Service General Hospital in Taipei from 2011 to 2015. All participants were genotyped for TLR4 rs10116253T/C and rs7873784G/C by the iPLEX Gold single nucleotide polymorphisms assay. Analysis of the conditional Cox proportional hazard model was used to test the association between TLR4 polymorphisms and mortality. Results: Our results show that adjustment with age and gender, TLR4 rs10116253T/C demonstrates an association with mortality (hazard ratio [HR] =1.38, 95% confidence interval [CI] =1.00–1.89) in a dominant model, especially in infection related mortality (HR = 2.14, 95% CI = 1.04–4.43) of HD patients. Conclusion: The results demonstrate that TLR4 rs10116253T/C may play a role in the etiology of infection among HD patients.