多重聚合酶链反应快速检测嗜水气单胞菌和爱德华菌

目的根据嗜水气单胞菌株和爱德华菌株16S rDNA基因的结构特点,设计合成了二对引物XZAH3、XZAH4和XZE7b、XZE8,建立了一种同时检测鉴别嗜水气单胞菌株和爱德华菌株的多重PCR技术。试验结果表明,用这两对引物对嗜水气单胞菌和爱德华菌株进行多重PCR,嗜水气单胞菌株只扩增出361bp一条带,而爱德华菌株只扩增出576bp一条带,而对其他鱼病病原的扩增不出现任何条带,结果均为阴性;敏感性测定结果表明,该多重PCR最低能检出10pg的嗜水气单胞菌株、爱德华菌株的DNA模板。

Rapid defection Aeromonas hydrophila and Edwardsiella by multiplex polymerase chain reaction

The multiplex polymerase chain reaction(PCR) was developed and optimized to detect and identify Aeromonas hydrophila and Edwardsiella simultaneously.Two pairs of specific primers XZAH3,XZAH4 and XZE7b,XZE8 were designed according to the sequences of 16S rDNA of Aeromonas hydrophila and Edwardsiella.The results showed that only 361pb-long DNA fragment for the Edwardsiella strains was amplified and the 576pb-long DNA fragments for the Aeromonas hydrophila were amplified by the multiplex PCR with these two pairs of primers,but the other fish pathogenic viruses and bacteria were not amplified.As little as 10 pg DNA of Aeromonas hydrophila and Edwardsiella could be detected by the multiplex PCR.