| According to the gene sequences of hlyA gene in GenBank, one pairs of specific primer was designed for amplifying the specific fragments of hlyA gene. After optimization of annealing temperature and primers concentrations, PCR was established for simultaneous detection of the hlyA gene. The specific band of 600 bp was amplified. The sensitivity and specificity tests showed that the PCR was highly sensitive in 0.4 ng/L DNA. No band was amplified from nonpathogenetic A. hydrophila, E. coil, Flavobacterium, Citrobacter freundii by PCR. 123 clinical samples were detected by the PCR, conventional microbiology methods and their coherence was 97.6%. The results revealed that the established PCR assay was sensitive, specific and it could be used to detect pathogenetic A. hydrophila rapidly. |
