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目的研究secA1基因在诺卡菌菌种的鉴定分型能力。方法对59株诺卡菌的secA1基因进行聚合酶链反应扩增,扩增片段纯化后测序,并从Gen Bank数据库下载32株诺卡菌secA1基因序列进行补充,采用DNAStar软件对所有序列计算种内与种间相似度水平,并用Mega 6.06软件的邻接法构建诺卡菌菌种系统发育进化树。结果 91株受试诺卡菌secA1基因的种内相似度为97.2%~100.0%,平均相似度达98.6%;种间相似度为85.2%~98.4%,平均相似度达91.8%。以secA1基因构建的系统发育进化树中,16种诺卡菌被准确分离为16个独立的分支,诺卡菌近缘种新星诺卡菌复合体、少食/短链诺卡菌复合体均可有效分离。结论 secA1基因序列分析为诺卡菌菌种的鉴定分型提供了一种新方法,在诺卡菌近缘种的分型方面有独特的优势。
Objective To investigate the genotyping ability of secA1 gene in Nocardia species. Methods The secA1 gene of 59 strains of Nocardia was amplified by polymerase chain reaction (PCR) and sequenced. The sequence of secA1 gene of 32 strains of Nocardia was downloaded from the Gen Bank database. DNA sequence analysis was performed using DNAStar software. Within and between species similarity level, and use Mega 6.06 software adjacency method to construct the Neocarray phylogenetic evolution tree. Results The intraspecies similarity of secA1 gene from 91 strains tested was 97.2% -100.0% and the average similarity was 98.6%. The similarity between species was 85.2% -98.4%, and the average similarity was 91.8%. In the phylogenetic tree constructed with the secA1 gene, 16 kinds of Nocardia were accurately separated into 16 independent branches. The Nocardia nematocyst Nocardia complex and the Eat / Short Nocardia complex were Effective separation. Conclusion The sequence analysis of secA1 gene provides a new method for the identification and identification of the genus Nocardia. It has unique advantages in the genotyping of Nocardia related species.