The method of Salmonella detection recommended is cultural, but it is laborious, presents a high consumption of material, and requires about five days for presumptive results. Immunosensor is an alternative tool that has shown promising and rapid results, although many devices have their performance evaluated only under buffering conditions and few achieve the validation stage. The objective was to perform a pre-collaborative validation of an electrochemical immunosensor assembled on screen-printed electrodes for the detection of Salmonella sp. in milk. The antibodies were immobilized by cysteamine self-assembled monolayer. The sandwich-type amperometric immunosensor was evaluated for contaminated raw and whole UHT milk and compared to performance with a gold standard reference method (BAM) according to AOAC recommendations for a single laboratory. A binary response (positive/negative) of the immunosensor was used based on a cut off established from current electric obtained for the absence of the pathogen. There was no significant difference for the results of the biosensor and the reference method, in the absence and the levels from 10¹ to 10³ CFU mL⁻¹ of Salmonella Typhimurium for the two types of milk. This result indicates the efficiency of the biosensor in detecting the pathogen into a complex matrix.