Detection of Edwardsiella by two-temperature PCR

One pair of primers were designed and synthesized according to the published sequence of the 16S rDNA of Edwardsiella.A two-temperature polymerase chain reaction method was developed for detection of Edwardsiella.DNAs isolated from six Edwardsiella strains were amplified by the two-temperature PCR,leading to the PCR products of 576 bp.But the other 10 pathogens,such as Aermonas hydrophila,Aermons sobria,Pseudomonas fluorescens,Cytophaga columnaris,Streptococcus,Staphylococci,Vibrio,Escherichia colibacillus and Salmonella,failed to show any positive results.It was found that as little as 1 pg of Edwardsiella DNA and 48 bacteria were sufficient to be detected by this method.This two-temperature PCR provides a rapid diagnostic method for early identification and control of Edwardsiella in the field of aquatic food safety.