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Toll-like receptors (TLR) play an eminent role in the regulation of immune responses to invading pathogens during sepsis. TLR genetic variants might influence individual susceptibility to developing sepsis. The current study aimed to investigate the association of genetic polymorphisms of the <i>TLR2</i> and <i>TLR4</i> with the risk of developing sepsis with both a pilot study and in silico tools. Different in silico tools were used to predict the impact of our SNPs on protein structure, stability, and function. Furthermore, in our prospective study, all patients matching the inclusion criteria in the intensive care units (ICU) were included and followed up, and DNA samples were genotyped using real-time polymerase chain reaction (RT-PCR) technology. There was a significant association between <i>TLR2</i> Arg753Gln polymorphisms and sepsis under the over-dominant model (<i>p</i> = 0.043). In contrast, we did not find a significant difference with the <i>TLR4</i> Asp299Gly polymorphism with sepsis. However, there was a significant association between <i>TLR4</i> Asp299Gly polymorphisms and <i>Acinetobacter baumannii</i> infection which is quite a virulent organism in ICU (<i>p</i> = 0.001) and post-surgical cohorts (<i>p</i> = 0.033). Our results conclude that the <i>TLR2</i> genotype may be a risk factor for sepsis in adult patients.