Development of a multiplex PCR assay for the identification of Salmonellaenterica serovarenteritidis

In order to develop a rapid method for the detection of the prevalent Salmonella enterica serovar enteritidis,a multiplex PCR method was developed and validated.Portions of the gene sequences of somatic antigen for Salmonella serogroups A/D1,flagellar antigens for fliC-HG and Salmonella difference fragment(sdfI) were targeted for amplification using four primer pairs.The multiplex PCR was developed and optimized.To valid the assay,genomic DNA from 14 Salmonella strains representing 14 serotypes and 18 non-salmonella strains was subjected to the PCR.The method was applied to the detection of 53 samples isolated from 2009 to 2010 in Zhejiang Province.The results showed the four targeted genes were correctly amplified only from Salmonella enteritidis.The assay could differentiate Salmonella serogroup A from Salmonella serogroup D1,and it also could identify the Salmonella with fliC-HG.The coincidence rate of the actual sample detection was up to 100%.It is concluded that this multiplex PCR method can identify Salmonella enteritidis enterica serovars directly.It can make up for deficiencies of uneven quality of commerical serum.It should be an assistant method to the traditional serotyping method.It is a simple,rapid,reliable and reproducible method of Salmonella enteritidis detection that will aid in surveillance,prevention and control of this pathogen.