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Detection of C ronobacter on glu B Gene and Differentiation of Four C ronobacter Species by Polymerase Chain Reaction‐Restriction Fragment Length Polymorphism Typing
Author(s) -
Ye Yingwang,
Ling Na,
Han Yongjia,
Cao Xiaodong,
Wu Qingping
Publication year - 2015
Publication title -
journal of food safety
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.427
H-Index - 43
eISSN - 1745-4565
pISSN - 0149-6085
DOI - 10.1111/jfs.12192
Subject(s) - restriction fragment length polymorphism , biology , taqi , polymerase chain reaction , typing , microbiology and biotechnology , terminal restriction fragment length polymorphism , restriction enzyme , gene , genetics
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.