16SrDNA序列分析在鉴定布鲁氏菌中的应用

Application of the 16S rDNA sequence analysis method in identification of Brucella

【目的】建立布鲁氏菌的16S rDNA序列分析方法,评价该方法鉴定布鲁氏菌的特异性和实用性。【方法】用PCR扩增布鲁氏菌的16S rDNA片段,将扩增的产物纯化后测序,从GenBank下载与布鲁氏菌易发生血清学交叉反应的细菌的16S rDNA序列。使用DNAMAN软件进16S rDNA序列相似性分析。【结果】在布鲁氏菌中16S rDNA核苷酸序列相似性达到了99.74%,而与其他有血清型交叉反应的菌株相比较,16S rDNA序列间有显著差异。【结论】16S rDNA序列分析是一种快速、简便、特异的鉴定布鲁氏菌的方法之一。

[Objective] To establish the 16S rDNA sequence analysis method to identifiy Brucella and evaluate the specificity and feasibility of this method. [Methods] 16S rDNA were amplified from Brucella by polymerase chain reaction （PCR） method and the purified product were directly sequenced for further analysis. The 16S rDNA sequence of the bacteria that are known to cross-react serologically with Brucella were downloaded from the GenBank. DNAMAN was used for comparison of 16S rDNA sequence. [Results] Brucella 16S rDNA sequence similarity reached 99.74%, Brucella 16S rDNA sequence to serologically related bacteria had more pronounced differences. [Conclusion] 16S rDNA sequence analyses is a rapid, simple diagnostic and specific method.