期刊
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
卷 78, 期 11, 页码 3966-3974出版社
AMER SOC MICROBIOLOGY
DOI: 10.1128/AEM.07825-11
关键词
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资金
- National 973 Program of China [2012CB721001, 2012CB721101, 2011CB504900]
- National Natural Science Foundation of China (NSFC) [31030002, 31000044, 81171524]
- National 863 Program of China [2011BAK10B02]
- Fundamental Research Funds for the Central Universities [65010721, 65020121, 65020061]
- Research Fund for the Doctoral Program of Higher Education of China [20090031120023]
- National Key Programs for Infectious Diseases of China [2009ZX10004-108]
The Gram-negative bacterium Cronobacter sakazakii is an emerging food-borne pathogen that causes severe invasive infections in neonates. Variation in the O-antigen lipopolysaccharide in the outer membrane provides the basis for Gram-negative bacteria serotyping. The O-antigen serotyping scheme for C. sakazakii, which includes seven serotypes (O1 to O7), has been recently established, and the O-antigen gene clusters and specific primers for three C. sakazakii serotypes (O1, O2, and O3) have been characterized. In this study, the C. sakazakii O4, O5, O6, and O7 O-antigen gene clusters were sequenced, and gene functions were predicted on the basis of homology. C. sakazakii 04 shared a similar O-antigen gene cluster with Escherichia coli O103. The general features and anomalies of all seven C. sakazakii O-antigen gene clusters were evaluated and the relationship between O-antigen structures and their gene clusters were investigated. Serotype-specific genes for O4 to O7 were identified, and a molecular serotyping method for all C. sakazakii O serotypes, a multiplex PCR assay, was developed by screening against 136 strains of C. sakazakii and closely related species. The sensitivity of PCR-based serotyping method was determined to be 0.01 ng of genomic DNA and 10(3) CFU of each strain/ml. This study completes the elucidation of C. sakazakii O-antigen genetics and provides a molecular method suitable for the identification of C. sakazakii O1 to O7 strains.
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