通过rpoB序列分析和多重PCR鉴定炭疽芽孢杆菌。
Identification of Bacillus anthracis by rpoB sequence analysis and multiplex PCR.
作者信息
Ko Kwan Soo, Kim Jong-Man, Kim Jong-Wan, Jung Byeong Yeal, Kim Wonyong, Kim Ik Jung, Kook Yoon-Hoh
机构信息
Department of Microbiology and Cancer Research Institute, Institute of Endemic Diseases, SNUMRC, Seoul National University College of Medicine, Seoul National University Hospital, Seoul 110-799, Korea.
出版信息
J Clin Microbiol. 2003 Jul;41(7):2908-14. doi: 10.1128/JCM.41.7.2908-2914.2003.
Comparative sequence analysis was performed upon Bacillus anthracis and its closest relatives, B. cereus and B. thuringiensis. Portions of rpoB DNA from 10 strains of B. anthracis, 16 of B. cereus, 10 of B. thuringiensis, 1 of B. mycoides, and 1 of B. megaterium were amplified and sequenced. The determined rpoB sequences (318 bp) of the 10 B. anthracis strains, including five Korean isolates, were identical to those of Ames, Florida, Kruger B, and Western NA strains. Strains of the "B. cereus group" were separated into two subgroups, in which the B. anthracis strains formed a separate clade in the phylogenetic tree. However, B. cereus and B. thuringiensis could not be differentiated. Sequence analysis confirmed the five Korean isolates as B. anthracis. Based on the rpoB sequences determined in the present study, multiplex PCR generating either B. anthracis-specific amplicons (359 and 208 bp) or cap DNA (291 bp) in a virulence plasmid could be used for the rapid differential detection and identification of virulent B. anthracis.
对炭疽芽孢杆菌及其亲缘关系最近的蜡样芽孢杆菌和苏云金芽孢杆菌进行了比较序列分析。对来自10株炭疽芽孢杆菌、16株蜡样芽孢杆菌、10株苏云金芽孢杆菌、1株蕈状芽孢杆菌和1株巨大芽孢杆菌的rpoB DNA片段进行了扩增和测序。包括5株韩国分离株在内的10株炭疽芽孢杆菌菌株的rpoB序列（318 bp）与Ames、佛罗里达、克鲁格B和北美西部菌株的序列相同。“蜡样芽孢杆菌群”菌株被分为两个亚群，其中炭疽芽孢杆菌菌株在系统发育树中形成一个单独的分支。然而，蜡样芽孢杆菌和苏云金芽孢杆菌无法区分。序列分析证实这5株韩国分离株为炭疽芽孢杆菌。基于本研究确定的rpoB序列，产生炭疽芽孢杆菌特异性扩增子（359和208 bp）或毒力质粒中的cap DNA（291 bp）的多重PCR可用于快速鉴别检测和鉴定有毒力的炭疽芽孢杆菌。
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