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Title: Molecular typing of isolates of the fish pathogen, Flavobacterium columnare, by single-strand conformation polymorphism analysis

Author
item OLIVARES-FUSTER, OSCAR - AUBURN UNIVERSITY
item Shoemaker, Craig
item Klesius, Phillip
item ARIAS, COVADONGA - AUBURN UNIVERSITY

Submitted to: FEMS Immunology and Medical Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/30/2007
Publication Date: 5/20/2007
Citation: Olivares-Fuster, O., Shoemaker, C.A., Klesius, P.H., Arias, C.R. 2007. Molecular typing of isolates of the fish pathogen, Flavobacterium columnare, by single-strand conformation polymorphism analysis. FEMS Immunology and Medical Microbiology 269: 63-69.

Interpretive Summary: Flavobacterium columnare is a gram negative bacterial pathogen of channel catfish responsible for millions of dollars in annual losses to the commercial catfish industry in the United States. Previous work has documented the genetic diversity of F. columnare. This study used another method (singe strand sequence conformation (SSCP)) that was faster, easier and less costly than previously described genotyping methods (such as DNA sequencing). This technique (SSCP) can be used in epidemiology studies to detect genetic diversity of F. columnare isolated from fish farms and the environment.

Technical Abstract: Flavobacterium columnare intraspecies diversity was revealed by analyzing the 16S rRNA gene and the 16S-23S internal spacer region (ISR). Standard restriction fragment length polymorphism (RFLP) of these sequences was compared with single strand conformation polymorphism (SSCP). Diversity indexes showed that both 16S-SSCP and ISR-SSCP improved resolution (D greater than or equal to 0.9) when compared with standard RFLP. ISR-SSCP offered a simpler banding pattern than 16S-SSCP while providing high discrimation between isolates. SSCP analysis of rRNA genes proved to be a simple, rapid and cost effective method for routine fingerprinting of F. columnare.