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Selection of Universal Primers for PCR Quantification of Total Bacteria Associated With Fish Fillets

An optimal universal primer pair DG74/RW01 for quantitative PCR detection of the mixed bacterial flora from fish fillets was selected from five pairs of primers. The minimum detection limits with GelStar™ and ethidium bromide (EB) used as agarose stains for DNA bands with these primers were found to be 5 and 1 × 102 CFU/PCR, respectively. The liner range of DNA amplification was from 50 to 1 × 105 with GelStar™ and from 5 × 102 to 1 × 105 with EB.