爱德华氏菌常规PCR检测体系的建立  

Development of a conventional PCR method for the detection of Edwardsiella ictaluri

作　　者：刘天强 黄冠军 刘衍鹏 肖丹 

机构地区：[1]通威股份有限公司动物保健研究所,四川成都610041

出　　处：《淡水渔业》2013年第4期76-79,共4页Freshwater Fisheries

基　　金：四川省科技支撑计划项目(2011NZ0071)

摘　　要：根据NCBI公布的爱德华氏菌(Edwardsiella ictaluri)磷酸丝氨酸转氨酶(phosphoserine transaminase,serC)的基因序列,设计一对特异性引物,建立了可快速而准确地检测爱德华氏菌的PCR方法,并对患病鱼组织进行了检测。研究结果表明,使用设计的引物能够扩增出与预计大小一致的124 bp的特异性片段,具有较好的检测特异性,对靶标DNA的检测灵敏度为50 pg/反应,对靶标细菌的检测灵敏度为56 cfu/反应。样品的检测结果与实际发病情况一致,表明成功建立了爱德华氏菌常规PCR检测方法,可用于爱德华氏菌病的诊断。A pair of PCR primers was designed according to the gene sequence of the phosphoserine transaminase gene （GenBank accession number： AF110153.1 ） of Edwardsiella ictaluri and was used to establish the PCR method for detection of E. ictaluri. Samples collected from infected fish were detected with the developed PCR and the results showed that the expected product with a size of 124bp was amplified by the PCR assay. The specificity of the established PCR method was 50 pg/reaction for the genomic DNA of E. ictaluri and 56 cfu/reaction for pathogen. The result of the detection was in conformity with actual case of the infectious diseases, which indicated that the PCR diagnostic method had been established successfully and could be used to detect E. ictaluri in infected samples.

关 键 词：鮰爱德华氏菌(Edwardsiella ictaluri) 磷酸丝氨酸转氨酶 聚合酶链式反应 检测 

分 类 号：S943.199.42[农业科学—水产养殖]