Detection of C ronobacter on glu B Gene and Differentiation of Four C ronobacter Species by Polymerase Chain Reaction‐Restriction Fragment Length Polymorphism Typing

C ronobacter species are foodborne pathogens associated with neonatal meningitis, sepsis and necrotizing enterocolitis through consumption of contaminated powdered milk. However, there are no specific targets used for differentiating C ronobacter species. The DNA ‐based assay and polyphasic analysis have facilitated the detection and typing of C ronobacter species in foodborne outbreaks. In this study, polymerase chain reaction ( PCR ) and dot hybridization for detecting C ronobacter spp. was developed targeting glu B gene, then one PCR ‐restriction fragment length polymorphism ( RFLP ) typing assay with TaqI digestion was applied for four species differentiation within C ronobacter . Results indicated that the glu B gene is a specific target for detecting C ronobacter and the PCR‐RFLP typing assay could conveniently and rapidly differentiate C ronobacter malonaticus , C . dublinensis , C . sakazakii and C . muytjensii . Practical Applications In this study, polymerase chain reaction ( PCR ) and dot hybridization targeting glu B for detecting C ronobacter was developed and a PCR ‐restriction fragment length polymorphism ( RFLP ) typing assay with TaqI was successfully applied for differentiating four C ronobacter species. Results indicated that glu B is a specific marker for detection of C ronobacter and PCR‐RFLP targeting glu B gene product digested with TaqI could conveniently differentiate four C ronobacter species.