Toll-like receptors (TLRs), the building blocks of the innate immune response, are largely responsible for the development of otitis media. The illness may worsen due to mistakes or faults in TLR expression in immune cells found in the blood or middle ear tissue. People with various bacterial infections and chronic otitis media were evaluated for their TLR (TLR2, TLR4, and TLR5) gene expression. The results showed that higher expression levels of TLR2 were associated with Staphylococcus aureus, Streptococcus pneumoniae, and Pseudomonas aeruginosa in mucosal cells, whereas lower expression levels were associated with Moraxella catarrhalis and Haemophilus influenzae. The study found that TLR4 gene expression was higher in the mucosal cells of patients infected with Moraxella catarrhalis, Pseudomonas aeruginosa, Haemophilus influenzae and Streptococcus pneumoniae, compared to healthy controls, while Staphylococcus aureus was associated with lower expression. The study found that patients with infections caused by Haemophilus influenzae, Moraxella catarrhalis, and Staphylococcus aureus had no significant expression levels of the TLR5 gene in their mucosa cells, while patients with infections caused by Pseudomonas aeruginosa and Streptococcus pneumoniae had higher expression levels. TLRs2 mRNA expression levels increased in patients with chronic otitis media infected with Gram-positive bacteria (Staphylococcus aureus and Streptococcus pneumoniae), while TLR-4 and TLR-5 levels increased in patients infected with Gram-negative bacteria (Pseudomonas aeruginosa and Streptococcus pneumoniae).