基于 Vntr 位点变异性分析的布鲁氏菌菌株区分

E. Anisimova, E. Dodonova, D. Mirgazov, Lenar Zaynullin, Konstantin Osyanin
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引用次数: 0

摘要

本研究的目的是评估使用已开发的 MLVA 方案区分布鲁氏菌病致病菌株的有效性。该方案包括使用改良的 MLVA 引物分析 15 个 VNTR 位点。为了对建议的 MLVA 方案进行体外测试,我们使用了之前从犬科布氏杆菌 RM 6/66、鼠疫布氏杆菌 1330、鼠疫布氏杆菌 183-L 和梅毒布氏杆菌 1565 株分离的 DNA。通过 PCR 进行 MLVA,然后在琼脂糖凝胶中分离扩增子。在 15 个 VNTR 位点中,有 10 个位点的扩增呈阳性,即 Bru6、Bru7、Bru9、Bru16 和 Bru18、Bru19、Bru21、Bru30、Bru43 和 Bru45。这些基因位点的分子大小已在 B. canis RM 6/66 和 B. suis 1330 的参考菌株中得到证实。GenBank 数据库中代表的菌株的 MLVA 结果也一并列出。通过搜索 NCBI 资源数据库,我们获得了犬布鲁氏菌、鼠布鲁氏菌、鲍氏布鲁氏菌和梅利特斯布鲁氏菌中 49 株布鲁氏菌的基因组序列。通过生物信息学分析,确定了这些菌株的十个 VNTR 位点的分子量和重复次数。根据 MLVA 的结果,构建了树枝图。根据对十个可变位点序列的系统进化分析,确定了所研究的大多数布鲁氏菌菌株按照其分类位置分布在树枝图上。因此,我们得出结论,我们提出的 MLVA 方案有可能用于布鲁氏菌菌株的区分。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
BRUCELLA STRAINS DIFFERENTIATION BASED ON VNTR LOCI VARIABILITY ANALYSIS
The purpose of the study is to evaluate the effectiveness of using the developed MLVA protocol for differentiating strains of the causative agent of brucellosis. This protocol includes the analysis of 15 VNTR loci using modified MLVA primers. For in vitro testing of the proposed MLVA scheme, we used previously isolated DNA from strains B. canis RM 6/66, B. suis 1330, B. suis 183-L, B melitensis 1565. MLVA was carried out by PCR followed by separation of amplicons in an agarose gel. Positive amplification was observed for 10 of the 15 VNTR loci, namely Bru6, Bru7, Bru9, Bru16 and Bru18, Bru19, Bru21, Bru30, Bru43 and Bru45. The molecular size of these loci for the reference strains B. canis RM 6/66 and B. suis 1330 was confirmed in silico. MLVA results for strains represented in the GenBank database are also presented. By searching the NCBI resource databases, we obtained the genomic sequences of 49 Brucella strains of the species B. canis, B. suis, B. aborus, and B melitensis. Using bioinformatic analysis, the molecular weight of each of the ten VNTR loci and the number of repeats in it were determined for these strains. Based on the results of the MLVA, a dendrogram was constructed. Based on a phylogenetic ana¬lysis of the sequences of ten variable loci, it was established that the majority of the studied Brucella strains were distributed on the dendrogram in accordance with their taxonomic position. Thus, we concluded that our proposed MLVA protocol has the potential to be used for the differentiation of Brucella strains.
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