Title: Establishment of duplex PCR for detection of pathogenic Aeromonas schubertii
Abstract: To develop a rapid method to detect pathogenic Aeromonas schubertii in fish,a duplex PCR method was established with two pairs of primers designed according to the conservative sequences of gyrB and hlyA genes of A.schubertii,respectively.By optimization of PCR conditions,the results showed that two DNA fragments of 601 bp(gyrB) and 375 bp(hlyA) were amplified from the genomic DNA extracted from A.schubertii by the duplex PCR,with a limit detection of 2.3 ×10-2ng bacteria genomic DNA or 3 ×102cfu of A.schubertii.In addition,the specificity tests showed that there were no cross-reactions with other bacteria such as A.sobria,A.hydrophila,Streptococcus agalactiae,Nocardia seriolae,Edwardsiella tarda,Pseudomonas fluorescens,Flavobacterium columnare,Vibrio harveyi and Plesiomonas shigelloides.Moreover,the method was applied to detect clinical samples and the results showed 100% consistence with those of the conventional bacterial isolation and identification method.Therefore,the duplex PCR could be used in quick diagnose and epidemiology investigation of A.schubertii infection.
Publication Year: 2014
Publication Date: 2014-01-01
Language: en
Type: article
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Cited By Count: 2
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