Establishment and application of multiplex PCR detection method for Staphylococcus aureus,Streptococcus,Salmonella spp and Escherichia coli

A multiplex PCR assay was developed and evaluated for the clinical detection of Staphylococcus aureus,Streptococcus,Salmonella spp and Escherichia coli.Four pairs of primers were designed and synthesized according to highly conserved regions of the Streptococcus ef-tu gene,Staphylococcus aureus nuc gene,Salmonella spp hut gene and Escherichia coli 23SrRNA gene of NCBI.The multiplex PCR reaction condition was optimized and the multiplex PCR method for detecting the co-infection of Staphylococcus aureus,Streptococcus,Salmonella spp and Escherichia coli was established.The specificity test showed that a fragment of 197,278,495 and 652 bp was amplified from genomic DNA of Streptococcus,Staphylococcus aureus,Salmonella spp and Escherichia coli,respectively.No amplification was achieved from control groups of other bacteria.The sensitivity test showed that the multiplex PCR could detect genome DNA of 25.6 pg for Streptococcus,33.2 pg for Staphylococcus aureus,35.7 pg for Salmonella spp and 52.1pg for Escherichia coli.The artificial simulation test showed that the multiplex PCR could detect four kinds of pathogens from co-infection disease material.The result indicated the multiplex PCR method had advantages of specificity,sensitivity,repetitive,and it could effectively detect the co-infection of Streptococcus,Staphylococcus aureus,Salmonella spp and Escherichia coli in clinical samples.