Forschungs- & Entwicklungsinformationsdienst der Gemeinschaft - CORDIS

FP5

ASIARESIST Berichtzusammenfassung

Project ID: ICA4-CT-2001-10028
Gefördert unter: FP5-INCO 2
Land: United Kingdom

(GTG)5- PCR mediated typing of aquaculture-associated isolates

A repetitive DNA element PCR fingerprinting (rep-PCR) protocol was developed for the strain typing of aquaculture-associated bacterial isolates (n= 650). A simple boiled extract of broth cultures acted as a DNA template for the PCR.

The (GTG)5 primer was employed in combination with the convenience of a commercial PCR master mix and following amplification, the PCR samples separated under standardised electrophoresis conditions. Numerical analysis of the DNA profiles (GelCompar, Applied Maths, Belgium) allowed individual strain comparisons to be made. Replicates, isolates found to possess identical (GTG)5-PCR profiles originating within the same sample, could be revealed to yield a de-replicated strain set, thus reflecting the true genotypic diversity of the strain collection.

Analysis of the DNA profiles was also able to highlight the presence of possible strain types occurring between different samples. The identity of these PCR strain types was later confirmed by macrorestriction analysis, an independent typing technique. The rep-PCR typing method was found to be a suitable as a screening method, was simple to perform and could be universally applied to assess the genotypic variation of a strain collection.

Reported by

University of Stirling
Institute of Aquaculture, Pathfoot Building, University of Stirling
FK9 4LA Stirling
United Kingdom
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