Abstract: The aim of this study was the validation of the Micro Biological Survey (MBS) method for microbiological analysis of food for Total Viable Count (TVC) and Escherichia coli (E. coli). The MBS method is a rapid quantitative alternative method for the detection and selective counting of bacteria in agro-food, in water and in environmental samples. It is based on colorimetric survey in mono-use disposable reaction vials that must be filled with the samples without any preliminary treatment (e.g., homogenization, dilution, etc.); the greater the number of bacteria presents into the sample, the faster the color change. However, an independent evaluation of the analytical results obtained with MBS method would be required before commercialization. Therefore, this alternative method was validated in comparison to the reference method. The general estimate of precision, reliability, uncertainty, linearity, accuracy and selectivity were determined. All the performance parameters have demonstrated total correlation between the alternative method and the reference method for the detection and counting of TVC and E. coli both in artificially contaminated and in naturally contaminated samples. MBS assay can be used as rapid and user friendly screening method for detection of TVC and E. coli in food industry.