Objectives: To engage a multiplex PCR (mPCR) for identifying simultaneously Bacillus anthrads and Yersinia pestis from artificially infected environment samples. Materials and methods: B. anthacis Ba1 and Y. pestis Yp1 strains were inactivated at 121°C/20 minutes and then infected in soil, food, water samples. A previous optimized multiplex PCR for determining B. anthracis va Y. pestis were performed, positive PCR products were separately extracted from agarose gel and used for direct sequencing. Obtained sequences were analyzed by Bioedit and Geneious R7 softwares, and Blast tools. Results and conclusion: the authors succeed to establish models of artificially infected environment samples including soil, water and food. Both B. anthacis and Y. pestis were specifically determined by mPCR and confirmed by sequencing.