Isolation of an unusual strain of Edwardsiella tarda from turbot and establish a PCR detection technique with the gyrB gene
Isolation of an unusual strain of Edwardsiella tarda from turbot and establish a PCR detection technique with the gyrB gene
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大菱鲆特异爱德华氏菌的分离及gyrB基因PCR检测技术的建立
DOI：
10.1111/j.1365-2672.2008.03779.x
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发表时间：
2008-09-01
影响因子：
4
通讯作者：
Han, Y.
中科院分区：
文献类型：
--
作者：
Lan, J.;Zhang, X. -H.;Han, Y.
目的：本研究的目的是报告一种不寻常的迟缓爱德华氏菌，并开发一种有效的方法来鉴定这种细菌。方法和结果：2006年春夏期间，中国青岛养殖大菱鲆（Scophamus maximus）中发生了一起动物疫病流行。从受感染的鱼中分离出一种革兰氏阴性杆状细菌（命名为LTB-4），并被证明对大菱鲆具有毒性。根据16S rDNA测序和表型测试，细菌病原体被鉴定为E. tarda。与那些通常描述的 E. tarda 菌株不同，没有观察到鞭毛虫。使用gyrB基因的通用引物从E. tarda扩增部分gyrB基因并测序。 gyrB 基因的聚合酶链式反应 (PCR) 引物是专为 E. tarda 设计的。结果显示，E. tarda 中的 gyrB 片段呈阳性扩增，而其他细菌种类呈阴性。此外，该技术还实现了病鱼中E. tarda的识别。结论：该菌株被鉴定为无鞭毛的E. tarda，并以gyrB基因作为分类标记，建立了有效的E. tarda鉴定方法。 研究意义和影响：罕见的E. tarda在中国首次报道，PCR技术可以快速、灵敏地检测E. tarda。
Aims: The aim of this study was to report an unusual Edwardsiella tarda and develop an effective method to identify this bacterium.Methods and Results: During the spring and summer of 2006, an epizootic occurred among cultured turbot (Scophthalmus maximus) in Qingdao, China. A gram-negative, rod-shaped bacterium (designated as LTB-4) was isolated from the infected fish, and was proved to be virulent to turbot. Based on the 16S rDNA sequencing and phenotypic tests, the bacterial pathogen was identified as E. tarda. Unlike those commonly described E. tarda strains, no flagellate was observed. Partial gyrB genes were amplified from E. tarda using the universal primers of gyrB genes and sequenced. The polymerase chain reaction (PCR) primers for the gyrB gene were designed specific to E. tarda. It revealed positive amplification of the gyrB fragment in E. tarda, whereas other bacterial species were negative. In addition, the technique enabled the recognition of E. tarda from diseased fish.Conclusions: The isolate was identified as E. tarda without flagellate and an effective method was developed to identify E. tarda based on using the gyrB gene as a taxonomic marker.Significance and Impact of the Study: The unusual E. tarda was first reported in China and the PCR allowed the rapid and sensitive detection of E. tarda.