中华鳖病原菌菌种的分离鉴定及多重PCR的初步建立  

Isolation,Identification and the Preliminary Establishment of the Multiplex-PCR of Pathogens from Soft-shelled Turtle

作　　者：燕晶[1] 刘文青[1] 高珊珊[1] 赵娜[1] 张亚宁[1] 赵宝华[1] 

机构地区：[1]河北师范大学生命科学学院,河北石家庄050024

出　　处：《河北师范大学学报（自然科学版）》2013年第2期177-185,共9页Journal of Hebei Normal University：Natural Science

基　　金：河北省重大科技攻关项目(0760621234)

摘　　要：对患病中华鳖心脏、肝脏、肠道中分离得到的疑似致病菌株进行常规生理生化鉴定,同时采用分子生物学鉴定方法,设计细菌16srDNA保守区通用引物,进行基因扩增及序列比对,建立并优化多重PCR反应条件.分离鉴定出4种病原菌:迟钝爱德华氏菌(Edwardsiella tarda,Et)、嗜水气单胞菌(Aeromonas hydrophila,Ah)、维氏气单胞菌(Aeromonas veronii,Av)、弗氏柠檬酸杆菌(Citrobacter freundii,Cf).多重PCR检测体系敏感性与特异性良好,证明多重PCR是一种能快速准确鉴定多种中华鳖致病菌的方法,且操作简单、灵敏度高、稳定性和重复性好.We identified the suspected pathogenic bacteria isolated from heart,liver and intestinal tract of soft-shelled turtle through conventional physiological and biochemical identification techniques. Multiplex PCR,which used primers designed according to conserved sequences in bacterial 16s rDNA,was also used for pathogen identification by gene amplification and sequence alignment,and the reaction conditions of multiplex PCR were optimized. By sequencing, Edwardsiella tarda （Et）, Aeromonas hydrophila （Ah）, Aeromonas veronii （Av）,Citrobacter freundii （C f） were identified. The high sensitivity and good specificity,stability and repeatability proved that the multiplex PCR was practical to identify pathogens from softshelled turtles.

关 键 词：中华鳖 生理生化鉴定 多重PCR 迟钝爱德华氏菌 嗜水气单胞菌 维氏气单胞菌 弗氏柠檬酸杆菌 

分 类 号：S947.1[农业科学—水产养殖]