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Journal of Clinical Microbiology, October 1998, p. 2835-2843, Vol. 36, No. 10
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Restriction Fragment Length Polymorphism Analysis of Some Flagellin Genes of Salmonella enterica

Catherine Dauga,1,* Anna Zabrovskaia,1,2 and Patrick A. D. Grimont1

Unité des Entérobactéries, INSERM U389, Institut Pasteur, 75724 Paris, France,1 and Institut Pasteur, 197101 St. Petersburg, Russia2

Received 19 March 1998/Returned for modification 26 May 1998/Accepted 30 June 1998

Salmonellae often have the ability to express two different flagellar antigen specificities (phase 1 and phase 2). At the cell level, only one flagellar phase is expressed at a time. Two genes, fliC, encoding phase-1 flagellin, and fljB, encoding phase-2 flagellin, are alternatively expressed. Flagellin genes from 264 serovars of Salmonella enterica were amplified by two phase-specific PCR systems. Amplification products were subjected to restriction fragment length polymorphism (RFLP) analysis by using endonucleases HhaI and HphI. RFLP with HhaI and HphI yielded 64 and 42 different restriction profiles, respectively, among 329 flagellin genes coding for 26 antigens. The phase-1 gene showed 46 patterns with HhaI and 30 patterns with HphI. The phase-2 gene showed 23 patterns with HhaI and 17 patterns with HphI. When the data from both enzymes were combined, 116 patterns were obtained: 74 for fliC, 47 for fljB, and 5 shared by both genes. Of these combined patterns, 80% were specifically associated with one flagellar antigen and 20% were associated with more than one antigen. Each flagellar antigen was divided into 2 to 18 different combined patterns. In the sample of strains used, determination of the phase-1 and phase-2 flagellin gene RFLP, added to the knowledge of the O antigen, allowed identification of all diphasic serovars. Overall, the diversity uncovered by flagellin gene RFLP did not precisely match that evidenced by flagellar agglutination.

* Corresponding author. Mailing address: Unité des Entérobactéries, INSERM U389, Institut Pasteur, 28 rue du Dr. Roux, 75724 Paris, France. Phone: 33 1 40613357. Fax: 33 1 45688837. E-mail: cdauga{at}pasteur.fr.

Journal of Clinical Microbiology, October 1998, p. 2835-2843, Vol. 36, No. 10
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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