The challenge of increasing food production while maintaining environmental sustainability can be addressed by using biofertilizers such as Azospirillum, which can enhance plant growth and colonize more than 100 plant species. The success of this biotechnology depends on the amount of plant growth-promoting bacteria associated with the plant during crop development. However, monitoring bacterial population dynamics after inoculation requires time-consuming, laborious, and costly procedures. To address these issues, this study describes an effective electrochemical DNA biosensor to detect Azospirillum brasilense. The biosensor comprises a glassy carbon electrode modified with a nanocomposite based on carbon nanotubes and gold nanoparticles capped with 3-n-propylpyridinium chloride silsesquioxane, followed by the immobilization of a thiolated probe oligonucleotide that binds specifically to the A. brasilense genome (AZO