Evaluation and selection of tandem repeat loci for a Brucella MLVA typing assay.
Evaluation and selection of tandem repeat loci for a Brucella MLVA typing assay.
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DOI：
10.1186/1471-2180-6-9
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发表时间：
2006-02-09
期刊：
影响因子：
4.2
通讯作者：
Vergnaud G
中科院分区：
文献类型：
--
作者：
Le Flèche P;Jacques I;Grayon M;Al Dahouk S;Bouchon P;Denoeud F;Nöckler K;Neubauer H;Guilloteau LA;Vergnaud G
布鲁氏菌的种和生物群的分类依赖于表型特征，有时由于分型试剂缺乏标准化而在解释结果时遇到困难。此外，这种生物分型的分辨率是中等的，需要操纵的活代理。更有效的基于DNA的方法是必要的，这项工作探讨了适用于多位点可变数目串联重复序列分析（MLVA）分型和物种鉴定。80个串联重复位点预测为多态性的基因组序列分析的三个可用的布鲁氏菌基因组序列进行了测试的多态性基因分型21布鲁氏菌菌株（18个参考菌株代表的6个“经典”的物种和所有的生物群，以及3个海洋哺乳动物菌株目前被认为是两个新物种的成员）。MLVA数据有效地根据其物种和生物型对菌株进行聚类。为了实际应用，选择了保留该聚类的15个位点的子集，并将其应用于236株分离株的分型。使用该MLVA-15测定，生成的簇对应于布鲁氏菌属的经典生物分型方案。这15个标记被分为两组，一组包括8个用户友好的小卫星标记，具有良好的物种识别能力（面板1），另一组互补的7个微卫星标记，具有更高的区分能力（面板2）。MLVA-15检测可应用于大量布鲁氏菌菌株的自动或手动程序，并可作为经典生物分型方法的补充，甚至替代。MLVA是基于非感染性物质（DNA），而生物分型程序本身需要操纵活病原体，这一事实促进了这一点。所产生的数据可在专门的劳动力和退伍军人事务部网站查询。
The classification of Brucella into species and biovars relies on phenotypic characteristics and sometimes raises difficulties in the interpretation of the results due to an absence of standardization of the typing reagents. In addition, the resolution of this biotyping is moderate and requires the manipulation of the living agent. More efficient DNA-based methods are needed, and this work explores the suitability of multiple locus variable number tandem repeats analysis (MLVA) for both typing and species identification. Eighty tandem repeat loci predicted to be polymorphic by genome sequence analysis of three available Brucella genome sequences were tested for polymorphism by genotyping 21 Brucella strains (18 reference strains representing the six 'classical' species and all biovars as well as 3 marine mammal strains currently recognized as members of two new species). The MLVA data efficiently cluster the strains as expected according to their species and biovar. For practical use, a subset of 15 loci preserving this clustering was selected and applied to the typing of 236 isolates. Using this MLVA-15 assay, the clusters generated correspond to the classical biotyping scheme of Brucella spp. The 15 markers have been divided into two groups, one comprising 8 user-friendly minisatellite markers with a good species identification capability (panel 1) and another complementary group of 7 microsatellite markers with higher discriminatory power (panel 2). The MLVA-15 assay can be applied to large collections of Brucella strains with automated or manual procedures, and can be proposed as a complement, or even a substitute, of classical biotyping methods. This is facilitated by the fact that MLVA is based on non-infectious material (DNA) whereas the biotyping procedure itself requires the manipulation of the living agent. The data produced can be queried on a dedicated MLVA web service site.
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影响因子：
9.4
作者：
Fabre, M;Koeck, JL;Pourcel, C
通讯作者：
Pourcel, C
影响因子：
2.6
作者：
Verger, JM;Grayon, M;Grimont, F
通讯作者：
Grimont, F
影响因子：
4.2
作者：
Le Flèche P;Fabre M;Denoeud F;Koeck JL;Vergnaud G
通讯作者：
Vergnaud G
DOI：
10.1073/pnas.221575398
发表时间：
2002-01-08
影响因子：
11.1
作者：
DelVecchio, VG;Kapatral, V;Overbeek, R
通讯作者：
Overbeek, R
影响因子：
5.8
作者：
Cloeckaert, A;Verger, JM;Godfroid, J
通讯作者：
Godfroid, J