To identify the anaerobic sporeforming Bacillus in dairy products.MethodsFive strains of gram-positive anaerobic sporeforming Bacilli in dairy product were identified by three methods, including Biochemical identification, MALDI-TOF MS and 16S rDNA sequencing techniques. ResultsThe biochemical method could identify three strains, whereas MALDI-TOF MS and 16S rDNA sequencing techniques could identify all five Bacilli. The three methods gave the same identification results on only one of the five Bacilli. By contrast, MALDI-TOF MS and 16S rDNA methods gave the same identification results on four of the five Bacilli, indicating that the two methods are highly identical in identifying the Bacillus. However, biochemical method had big variation in comparison with the other two methods.ConclusionMALDI-TOF MS and 16S rDNA sequencing methods can be used as supplemental methods to the biochemical method in rapid identification of anaerobic sporeforming Bacillus.