Improved pulsed-field gel electrophoresis procedure for the analysis of Flavobacterium columnare isolates previously affected by DNA degradation

Esteban Soto Martinez; Michael Mauel; Mark Lawrence

doi:10.1016/j.vetmic.2007.10.001

Improved pulsed-field gel electrophoresis procedure for the analysis of Flavobacterium columnare isolates previously affected by DNA degradation

Esteban Soto Martinez, Michael Mauel, Mark Lawrence

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

Abstract

Flavobacterium columnare is a fresh water bacterium that causes columnaris diseases in over 36 fish species. Intra-species typing of F. columnare can be performed by pulsed-field gel electrophoresis (PFGE). However, this method is hampered by the degradation of chromosomal DNA in about 10% of strains. In the current study, DNA degradation problems caused by extracellular DNases were overcome by fixation of cells with formaldehyde prior to isolation. The results substantiate that after problems due to DNases are overcome, PFGE analysis is a reproducible highly discriminating epidemiological method for studying F. columnare isolates regardless of fish host.

Original language	English (US)
Pages (from-to)	207-212
Number of pages	6
Journal	Veterinary Microbiology
Volume	128
Issue number	1-2
DOIs	https://doi.org/10.1016/j.vetmic.2007.10.001
State	Published - Apr 1 2008
Externally published	Yes

Keywords

DNA degradation
Flavobacterium columnare
Formaldehyde fixation
PFGE

ASJC Scopus subject areas

Animal Science and Zoology
Microbiology
veterinary(all)

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title = "Improved pulsed-field gel electrophoresis procedure for the analysis of Flavobacterium columnare isolates previously affected by DNA degradation",

abstract = "Flavobacterium columnare is a fresh water bacterium that causes columnaris diseases in over 36 fish species. Intra-species typing of F. columnare can be performed by pulsed-field gel electrophoresis (PFGE). However, this method is hampered by the degradation of chromosomal DNA in about 10% of strains. In the current study, DNA degradation problems caused by extracellular DNases were overcome by fixation of cells with formaldehyde prior to isolation. The results substantiate that after problems due to DNases are overcome, PFGE analysis is a reproducible highly discriminating epidemiological method for studying F. columnare isolates regardless of fish host.",

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AU - Mauel, Michael

AU - Lawrence, Mark

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AB - Flavobacterium columnare is a fresh water bacterium that causes columnaris diseases in over 36 fish species. Intra-species typing of F. columnare can be performed by pulsed-field gel electrophoresis (PFGE). However, this method is hampered by the degradation of chromosomal DNA in about 10% of strains. In the current study, DNA degradation problems caused by extracellular DNases were overcome by fixation of cells with formaldehyde prior to isolation. The results substantiate that after problems due to DNases are overcome, PFGE analysis is a reproducible highly discriminating epidemiological method for studying F. columnare isolates regardless of fish host.

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KW - Formaldehyde fixation

KW - PFGE

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