16S rDNA序列在诺卡菌菌种鉴定中的价值研究

doi:10.3969/j.issn.1002-2694.2015.11.006

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摘要目的评价16S rDNA序列在诺卡菌菌种鉴定中的应用价值。方法对 13种49株诺卡菌(43株标准株和6株临床株)16S rDNA序列片段进行PCR扩增,将扩增片段进行纯化后测序,从GenBank上下载32株诺卡菌(包括星形诺卡菌,少食诺卡菌,短链诺卡菌等)及3株棒状杆菌(Corynebacterium,C.),3株弗兰克氏菌属(Frankia,F.),3株分支杆菌(Mycobacterium,M.),3株链霉菌(Streptomyces,S.)的16S rDNA序列。对所测得的诺卡菌序列及GenBank所报道的诺卡菌序列用DNASTAR程序分析处理,计算种内及种间相似性,用Mega6.06构建诺卡菌菌种系统进化树。结果在81条受试诺卡菌16S rDNA序列中,16S rDNA能将诺卡菌大致可分为11群,除了不能将亚种分离外,诺卡菌其他种属均能得到很好的分离。诺卡菌16S rDNA种内及亚种内相似性为99.1% ~ 100%;种间相似性在95.7% ~ 98.8%之间。结论16S rDNA序列分析是一种快速,简单,特异性较好的诺卡菌菌种鉴定方法,能将诺卡菌鉴定至种的水平。

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	李路茜
	张媛媛
	刘海灿
	谭晓罗
	李振军
	楼永良

关键词 ：诺卡菌, 16S rDNA序列, 鉴定

Abstract：The objective was to evaluate the value of 16S rDNA sequence analysis method for identifying Nocardia species. In this study, we determined 49 Nocardia strains amplified by polymerase chain reaction (PCR) and the purified product were directly sequenced for further analysis. In the meantime, 32 Nocardia 16S rDNA sequences from GenBank were downloaded for supplement. DNA STAR software was used to analyse selected sequences and calculated the percentage similarity of sequence pairs. A cluster analysis dendrogram was constructed by Mega 6.06 software. The 16S rDNA sequence among Nocardia species had highly polymorphism except Nocardia complex (N. Africana, N. veterana and N. kruczakiae) sequences were highly consistent. Nocardia 16S rDNA sequence similarity reached 99.1% to 100%; the similarity between sequence pairs was 95.7% to 99.8%. In conclusion, 16S rDNA sequence analysis is a rapid, simple and specific method, which could identify Nocardia strains to species level.

Key words： Nocardia 16S rDNA sequences identification

PACS:

R378

基金资助:863课题(No.2014AA021404),国家科技重大专项(No.2013ZX10004221),公益性卫生行业专项(No.201302006)联合资助

通讯作者: 李振军,Email: lizhenjun@icdc.cn;楼永良,Email: louyongliang2013@163.com

引用本文:

李路茜，张媛媛，刘海灿，谭晓罗，李振军，楼永良. 16S rDNA序列在诺卡菌菌种鉴定中的价值研究[J]. 中国人兽共患病学报, 2015, 31(11): 1017-1022. LI Lu-xi, ZHANG Yuan-yuan, LIU Hai-can, TAN Xiao-luo, LI Zhen-jun, LOU Yong-liang. Evaluation of 16S rDNA sequence method in identification of Nocardia species. Chinese Journal of Zoonoses, 2015, 31(11): 1017-1022.

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http://www.rsghb.cn/CN/10.3969/j.issn.1002-2694.2015.11.006 或 http://www.rsghb.cn/CN/Y2015/V31/I11/1017

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