Hearing loss can occur at different physiological levels and degrees, primarily affecting children. Law No. 12,303/2010, known as the ‘Otoacoustic Emissions Test Law,’ requires newborns to undergo auditory screening. Different non-calibrated sound instruments can be used to perform behavioral auditory screening: plastic harmonica, spoon tapping on a cup, rubber toys, drums, straw maracas, and gourd maracas, with the latter three being of indigenous origin. The objective of this study is to verify the effectiveness of an indigenous Amazonian regional kit in neonatal behavioral auditory screening in Belém do Pará. A quantitative descriptive study was conducted for this purpose. The evaluated auditory kit with regional instruments proved to be effective, assessing the newborn’s main reflexes. It also demonstrated the simplicity of a detection method and the feasibility of its inclusion in routine neonatal screening methods, enabling the early diagnosis and monitoring of deafness in newborns, which can be presented as an alternative, especially in locations that do not have traditional equipment.