Optimized ribotyping protocol applied to Hungarian Bordetella bronchiseptica isolates:: identification of two novel ribotypes
Optimized ribotyping protocol applied to Hungarian Bordetella bronchiseptica isolates:: identification of two novel ribotypes
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DOI：
10.1016/s0378-1135(99)00118-2
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发表时间：
1999-09-29
影响因子：
3.3
通讯作者：
Magyar, T
中科院分区：
文献类型：
--
作者：
Register, KB;Magyar, T
Rie先前报道，用PvuII和来自大肠杆菌rrnB基因的探针产生的核糖体分型模式可用于区分支气管败血波氏杆菌的分离株。在本研究中，我们报告了对原始核糖体分型程序的修改，允许在先前表征的分离株中检测到与rrnB具有同源性的另外8个片段。重新定义核糖型以包括这些片段。虽然这种修饰不允许从先前表征的分离株中检测到新的核糖体型，但它确实导致了这些分离株中的五种更准确地重新分类为其他现有的核糖体型。据推测，额外的片段可以在未来的分析中形成新的核糖体分型的基础，这得到了随后对101只先前未表征的猪、兔和犬B，支气管败血症分离株的评价的支持。从该组中总共检测到六种不同的模式，包括两种以前未鉴定的核糖体类型17和18。核糖体型17的谱包括与任何其他核糖体型不相关的新片段，构成核糖体型18的片段的子集，对于其与其他核糖体型的区分是必需的，仅在本文报道的修改的条件下是可检测的，匈牙利猪分离物是高度克隆的，因为98.2%被鉴定为核糖体型3。同样，来自匈牙利的83.7%的兔分离株也是核糖体型3，聚类分析显示，尽管存在许多核糖体型，但B。支气管败血症分离株显示有限的异质性。在本研究中描述的改良条件下检测额外核糖体分型的能力加强了核糖体分型作为流行病学工具的有用性。出版社：Elsevier Science B.V.
Rie reported previously that ribotype patterns generated with PvuII and a probe derived from the Escherichia coli rrnB gene could be used to differentiate isolates of Bordetella bronchiseptica, In the present study we report modifications made to the original ribotyping procedure that permit detection in the formerly characterized isolates of an additional 8 fragments with homology to rrnB. Ribotypes were redefined to include these fragments. Although this modification did not permit the detection of novel ribotypes from the previously characterized isolates, it did result in a more accurate reclassification of five of these isolates to other existing ribotypes. It was hypothesized that the additional fragments could form the basis for novel ribotypes in future analyses, and this was supported by the subsequent evaluation of 101 previously uncharacterized pig, rabbit, and dog B, bronchiseptica isolates from Hungary. A total of six different patterns were detected from this group, including two previously not identified that were designated ribotypes 17 and 18. The profile of ribotype 17 includes a novel fragment not associated with any other ribotype, A subset of the fragments constituting ribotype 18, essential for its differentiation from other ribotypes, is only detectable under the modified conditions reported here, Hungarian swine isolates are highly clonal, since 98.2% were identified as ribotype 3. Similarly, 83.7% of rabbit isolates from Hungary are also ribotype 3, Cluster analysis revealed that despite the existence of numerous ribotypes, B. bronchiseptica isolates display limited heterogeneity. The ability to detect additional ribotypes under the modified conditions described in this study strengthens the usefulness of ribotyping as an epidemiologic tool. Published by Elsevier Science B.V.