Through the treatment of the sodium deoxycholate to the cells,Ethidium bromide monoazide was utilized to selectively allow the PCR amplification of a targeted DNA sequence in viable but not dead cells of Listeria monocytogenes,namely SD-EMA-PCR.The optimal concentration of sodium deoxycholate incorporating into the PCR system could enhance the discrimination of viable and dead cells.The results showed that: with the treatment of the Sodium deoxycholate,the optimized light exposure time to achieve cross-linking of DNA by the EMA in dead cells and to photolyse the free EMA in solution was at least 25 min,and the maximum concentration of EMA which did not inhibit the PCR amplification of DNA extracted from viable cells was 9 μg/mL.but the minimum concentration of EMA which completely inhibited the PCR amplification of DNA extracted from dead cells was 1.6 μg/mL;a detection limit of the assay for the viable cells was 20 CFU/mL.SD-EMA-PCR could minimize the possibility of the false-positive by the way of EMA-PCR and reduce the cost of detection.It is a new efficient approach for the rapid detection of the isolate of Listeria monocytogenes.