The nested-PCR specificity was evaluated with purified genomic DNA from S. agalactiae ST991 and ATCC12386 strains as template, as well as total DNA obtained from fish muscle tissue inoculated withS. agalactiae ST991 and with genomic DNA extracted from pure Strep - tococcus pneumoniae, Enterococcus faecalis ATCC 49 619 29 272, Strep - tococcus iniae MY001, Lactococcus garviae MY001, and Aeromonas hydrophila KJ99 cultures. S. agalactiae ATCC 49 619 29 272 DNA (obtained from a pure bacterial culture) was serially diluted and each dilution was mixed with constant amounts of genomic DNA extracted from frozen or from paraffin-wax-embedded healthy tilapia brains to evaluate nested-PCR sensitivity in tissue.
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