Establishment of a Rapid PCR Assay for Aeromonas dhakensis
Aeromonas dhakensis(A.dhakensis)could cause septicaemia and organ inflammation in infected animals as a serious zoonotic pathogen.It is a precondition to prevent relevant diseases through accurate identification of the pathogen.In order to establish a rapid PCR method with high sensitivity and strong specificity for A.dhakensis,targeting at the genetic marker DK1 gene,corresponding primers were designed,and annealing temperature was optimized,followed by evaluation on the specificity and sensitivity of the method.The results showed that positive band(874 bp)of A.dhakensis could be amplified at the annealing temperature of 65℃by the primers Ad-f/Ad-r;the detection limit for A.dhakensis solution was 1.38 CFU/μL,and that for nucleic acids was 9.37×10-4 ng/μL;no specific bands were found by PCR assay taking 20 kinds of other aquatic pathogens(such as A.hydrophila and A.veronii)as templates.In conclusion,the established method,with strong specificity and high sensitivity,could be used to rapidly and accurately detect A.dhakensis from diseased fish and environmental samples,supporting clinical identification of the pathogen.
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