(鱼回)爱德华氏菌常规PCR检测体系的建立.pdfVIP

第43 卷第4 期 2013 年7 月 淡水渔业 Vol. 43 No.4 Freshwater Fisheries Ju12013 鱼回爱德华氏菌常规 PCR 检测体系的建立 刘天强，黄冠军，刘衍鹏，肖 丹 (通威股份有限公司动物保健研究所，四川成都 610041 ) 摘要:根据 NCBI 公布的细爱德华氏菌(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细爱德华氏菌的 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 峙的特异性片段，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，表明成功建立了细爱德华氏菌常规PCR 检测方法，可用于锢爱德华氏菌病的诊断。 关键词:细爱德华氏菌( Edwardsiella 比taluri) ; 磷酸丝氨酸转氨酶;聚合酶链式反应;检测 中图分类号: S943.199.42 文献标识码:A 文章编号: 1 ∞0-6907-( 2013 )04 -0076-04 Development of a conventional PCR method for the detection of Edwardsiella ictaluri LIU Tian-qiang , HUANG Guan-jun , LIU Yan-peng , XIAO Dan (AniTTU11 Health Research lnstitute 01 Tongwei C，ο. LTD , Chengdu 61ω41 ， China) Abstract: A pair of PCR primers was designed according to the gene sequence of the phosphoserine transaminase gene (GenBank accession number: AFl 10153. 1) of Edwardsiellα ictaluri and was used 10 establish the PCR method for detec- tion of E. ictaluri. Samples co!lected 丘。m infected fish were detected 有ith 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 con- formity with actual case of the infectious diseases , which indicated that the PCR diagnostic meth叫 had been establishe