Urdaci M C, Chakroun C, Faure D, Bernardet J F
Laboratoire de Microbiologie et Biochimie Appliquée, Ecole Nationale des Ingénieurs des Travaux Agricoles de Bordeaux, Gradignan, France.
Res Microbiol. 1998 Jul-Aug;149(7):519-30. doi: 10.1016/s0923-2508(98)80006-5.
A PCR-based method was developed to identify and detect Flavobacterium psychrophilum, the causative agent of "cold-water disease" and "rainbow trout fry syndrome" in salmonid fish. Two oligonucleotide primers were designed by comparing the 16S rRNA sequence of all taxa in the genus Flavobacterium and of representative species in most related genera within rRNA superfamily V. Purified chromosomal DNAs from all these bacterial species, from 25 F. psychrophilum isolates and from several other fish-pathogenic bacteria were used to assess the specificity of the reaction. Amplification products were generated only with F. psychrophilum DNA. The detection level, equivalent to approximately 10 to 100 bacterial cells, was increased 10-fold by hybridization with a radioactive probe. Preliminary experiments demonstrated that this procedure can also be applied to samples of infected tissue. This PCR assay is therefore a rapid, specific, and sensitive alternative to conventional plate culture methods for the identification and detection of F. psychrophilum.
已开发出一种基于聚合酶链反应（PCR）的方法，用于鉴定和检测嗜冷黄杆菌，它是鲑科鱼类中“冷水病”和“虹鳟鱼苗综合征”的病原体。通过比较黄杆菌属所有分类单元以及rRNA超家族V中最相关属的代表性物种的16S rRNA序列，设计了两种寡核苷酸引物。使用来自所有这些细菌物种、25株嗜冷黄杆菌分离株以及其他几种鱼类致病细菌的纯化染色体DNA来评估反应的特异性。仅用嗜冷黄杆菌DNA产生了扩增产物。通过与放射性探针杂交，检测水平提高了10倍，相当于约10至100个细菌细胞。初步实验表明，该方法也可应用于感染组织样本。因此，这种PCR检测方法是一种快速、特异且灵敏的替代传统平板培养方法，用于鉴定和检测嗜冷黄杆菌。
