应用二温式PCR检测诊断爱德华氏菌病

Detection of Edwardsiella by two-temperature PCR

根据爱德华氏菌(Edwardsiella)的16S rDNA基因序列设计一对特异性引物,用二温式PCR对6株爱德华氏菌均扩增出与预期大小相一致的576 bp产物,而对嗜水气单胞菌(Aeromonas hydrophila)、温和气单胞菌(Aeromonas sobria)、荧光假单胞菌(Pseudcmonas fluoroscercs)、柱状屈挠杆菌(Cytophaga columnaris)、链球菌(Streptococcus)、葡萄球菌(Staphylococci)、弧菌(Vibrio)、大肠杆菌(Escherichia colibacillus)和沙门氏菌(Salmonella)等10种病原体的扩增,结果全为阴性。该二温式PCR可以检测到1 pg的爱德华氏菌DNA模板和48个菌体。本实验建立的二温式PCR为爱德华氏菌病的早期诊断与有效的防治提供了快速检测方法,对水产品的食品安全有重要的意义。

One pair of primers were designed and synthesized according to the published sequence of the 16S rDNA of Edwardsiella. A two-temperature polymerase chain reaction method was developed for detection of Edwardsiella. DNAs isolated from six Edwardsiella strains were amplified by the two-temperature PCR, leading to the PCR products of 576 bp. But the other 10 pathogens, such as Aermonas hydrophila, Aermons sobria, Pseudomonas fluorescens, Cytophaga columnaris, Streptococcus, Staphylococci, Vibrio, Escherichia olibacillus and Salmonella, failed to show any positive results. It was found that as little as 1 pg of Edwardsiella DNA and 48 bacteria were sufficient to be detected by this method. This two-temperature PCR provides a rapid diagnostic method for early identification and control of Edwardsiella in the field of aquatic food safety.