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U.S. Department of Health and Human Services

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TITLE:  A restriction fragment length polymorphism-based polymerase chain reaction as an alternative to serotyping for identifying Salmonella serotypes
 
AUTHORS:  Hong Y;Liu T;Hofacre C;Maier M;White DG;Ayers S;Wang L;Maurer JJ;
 
YEAR:  2003
 
JOURNAL ABBREV:  Avian Dis
 
MONTH:  Apr
 
TYPE:  JOUR
 
REFMAN INDEX:  231
 
JOURNAL FULL:  Avian diseases
 
VOLUME:  47
 
ISSUE:  2
 
START PAGE:  387
 
END PAGE:  395
 
KEYWORDS:  analysis;Animals;classification;Dna;DNA Restriction Enzymes;Environment;Enzymes;Flagellin;Food;genetics;Georgia;isolation & purification;methods;microbiology;Polymerase Chain Reaction;Polymorphism,Restriction Fragment Length;Poultry;Poultry Diseases;Research;Salmonella;Salmonella Infections,Animal;Serotyping;veterinary;Veterinary Medicine;
 
ABSTRACT:  The phase 1 (fliC) and phase 2 (fljB) Salmonella flagella genes were analyzed by restriction fragment length polymorphism (RFLP)-polymerase chain reaction (PCR) to aid in the identification of different Salmonella serotypes. Twenty-four phase 1 flagellin and eight phase 2 flagellin genes could be differentiated among each other with restriction endonucleases Sau3A and HhaI in RFLP-PCR analysis. These flagellin genes comprise the major antigenic formulas for 52 serotypes of Salmonella sp., which include the common serotypes found in poultry and other important food animal species. With the knowledge of the O antigen composition determined from conventional O serotyping, 90% of the Salmonella serotypes could be identified by this double restriction enzyme RFLP analysis of fliC and fljB genes. This RFLP-PCR flagellar typing scheme was successfully applied to the identification of serotype for 112 Salmonella isolates obtained from poultry environment. There was a significant correlation between RFLP-PCR and conventional serotyping (chi-square, P < 0.001). Overall, PCR-RFLP proved to be a fast, accurate, and economical alternative approach to serotyping Salmonella sp
 
AFFILIATIONS:  Department of Avian Medicine, College of Veterinary Medicine, University of Georgia, Athens, GA 30602, USA
 
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