致病性嗜水气单胞菌气溶素基因PCR检测方法的建立

Detection Method of the Aerolysin Gene in Pathogenic Aeromonas hydrophila Based on Polymerase Chain Reaction

[目的]建立一种基于PCR的致病性嗜水气单胞菌的检测方法。[方法]采用平板划线法从患病鲫鱼病灶部位分离、纯化获得嗜水气单胞菌,采用CTAB法提取DNA,根据GenBank已经登录的致病性嗜水气单胞菌气溶素基因保守序列设计引物P1和P2,并以P1、P2为引物对其进行PCR扩增,将测序结果在Blast上进行比对分析。[结果]PCR扩增得到一条约540 bp的条带,Blast分析表明该基因与GenBank登录的气溶素基因的同源性达95.67%。[结论]试验建立的致病性嗜水气单胞菌气溶素基因PCR检测方法简单、可行。

[Objective] To establish a novel detection method of the Aerolysin gene in pathogenic Aeromonas hydrophila based on polymerase chain reaction（PCR）.[Method] The Aeromonas hydrophila were isolated from the infection focus of crucian with plate streaking.DNA was extracted by CTAB method.P1 and P2 primers were designed according to the conserved sequence of pathogenic Aeromonas hydrophila aerolysin gene registered on GenBank.The comparative analysis of sequencing results was performed by Blast method.[Result] DNA fragment of Aeromonas hydrophila was about 540 bp.Blast analysis showed that the homology between amplified gene and aerolysin gene logged GenBank was up to 95.67%.[Conclusion] The method is simple and feasible for detecting aerolysin gene in pathogenic Aeromonas hydrophila.