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微生物学通报 2011
Development of a PCR method for the detection of pathogenic Aeromonas veronii
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Abstract:
The aim of the study was explore a set of new detective targets in pathogenic Aeromonas veronii which was more specific than others at present,to develop a new duplex PCR method which could effectively detection the pathogenic A.veronii.Comparison analysis of the 16S rRNA gene se-quences of Aeromonas spp.was used to explore A.veronii specific targets,which were then evaluated and used to design specific primers,and select the conservative primers of aerolysin genes in Aeromo-nas spp.to explore pathogenic targets.PCR parameters were optimized,its reaction system was devel-oped,and its sensitivity and specificity were checked.12 Aeromonas stains and 10 non-Aeromonas stains were tested for pathogenic A.veronii by duplex PCR method.The duplex PCR can clearly iden-tify A.veronii from Aeromonas species,and can identify aerolysin-producing stains of A.veronii.The detection limit of genomic DNA by the duplex PCR was 1.35×10-3 mg/L.A novel duplex PCR method was successfully developed in this study,which could effectively detect pathogenic A.veronii with high accuracy and sensitivity.