Establishment and Application of Multiple PCR for Diagnosing Proteus mirabilis,Salmonella and Listeria monocytogenes

【Objective】 The aim of this study is to establish a multiplex polymerase chain reaction(PCR) assay for simultaneous identification of Proteus mirabilis,Salmonella and Listeria monocytogenes.【Method】Based on the gene sequences of positive adjustment factor R gene of Urease(ureR) in Proteus mirabilis,conservative invasive antigen gene(invA) in Salmonella and the hly gene in Listeria monoytogenes,a three pairs of primer were designed.The specificity and sensitivity of PCR were analyzed for single gene.Multiple PCR method has been developed after analysis and optimization reaction condition by orthogonal experimental design L16(43).【Result】 Aiming to Proteus mirabilis,Salmonella and Listeria monocytogenes,under the optimized conditions,the anticipated PCR products were 374 bp,724 bp and 215 bp,respectively,and the specificity was high,and the minimum detection limit was 105 CFU/mL,which was a dilute degree lower in the sensitivity of single PCR.The results were stable in simulated examination and in marketing examination.【Conclusion】A rapid,specific and sensitive multiplex PCR system has been established and it is valuable for diagnosis and monitoring for Proteus mirabilis,Salmonella and Listeria monocytogenes.