title: Duplex Real-Time PCR Method for the Differentiation of Cronobacter sakazakii and Cronobacter malonaticus.
abstract: Cronobacter sakazakii and Cronobacter malonaticus are the most common species of Cronobacter , so it is necessary to detect the two species as soon as possible in surveillance programs. We developed a real-time PCR method for identifying C. sakazakii and C. malonaticus from the genus Cronobacter . In this study, the two pairs of primers and probes were designed, targeting 16S rRNA and fusA, respectively. The specificity of the real-time PCR assay was validated with 112 strains of Cronobacter , including 56 C. sakazakii , 32 C. malonaticus , 16 Cronobacter dublinensis , 6 Cronobacter turicensis , and 2 Cronobacter muytjensii . The results showed that C. sakazakii and C. malonaticus were all correctly identified, consistent with the results of another method by analyzing the clustering of the fusA sequence. The detection limit for pure culture was 102CFU/ml and 103CFU/g for artificially contaminated rehydrated powdered infant formula. Therefore, the developed real-time PCR was a rapid, sensitive, and reliable method for the identification of C. sakazakii and C. malonaticus .
authors.affiliation: National Institute for Communicable Disease Control and Prevention and State Key Laboratory for Infectious Disease Prevention and Control, Chinese Center for Disease Control and Prevention, Beijing 102206, People's Republic of China.|Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education and School of Life Sciences, Jilin University, Changchun 130012, People's Republic of China.
authors.forename: Xiaofang
authors.initials: X
authors.lastname: Li
authors.affiliation: National Institute for Communicable Disease Control and Prevention and State Key Laboratory for Infectious Disease Prevention and Control, Chinese Center for Disease Control and Prevention, Beijing 102206, People's Republic of China.
authors.forename: Jinghua
authors.initials: J
authors.lastname: Cui
authors.affiliation: National Institute for Communicable Disease Control and Prevention and State Key Laboratory for Infectious Disease Prevention and Control, Chinese Center for Disease Control and Prevention, Beijing 102206, People's Republic of China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Du
authors.affiliation: National Institute for Communicable Disease Control and Prevention and State Key Laboratory for Infectious Disease Prevention and Control, Chinese Center for Disease Control and Prevention, Beijing 102206, People's Republic of China.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Cui
authors.affiliation: Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education and School of Life Sciences, Jilin University, Changchun 130012, People's Republic of China.
authors.forename: Yibing
authors.initials: Y
authors.lastname: Huang
authors.affiliation: National Institute for Communicable Disease Control and Prevention and State Key Laboratory for Infectious Disease Prevention and Control, Chinese Center for Disease Control and Prevention, Beijing 102206, People's Republic of China.
authors.forename: Biao
authors.initials: B
authors.lastname: Kan
mesh_terms: D059126:Cronobacter / Q000145:classification*; D044083:Cronobacter sakazakii / Q000145:classification*; D006801:Humans; D041943:Infant Formula; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D012336:RNA, Ribosomal, 16S
keywords: Cronobacter malonaticus; Cronobacter sakazakii; Duplex real-time PCR; Sensitivity; Specificity