通过对质粒pXO1和pXO2以及染色体DNA上的序列进行多重PCR分析来鉴定和表征炭疽芽孢杆菌。
Identification and characterization of Bacillus anthracis by multiplex PCR analysis of sequences on plasmids pXO1 and pXO2 and chromosomal DNA.
作者信息
Ramisse V, Patra G, Garrigue H, Guesdon J L, Mock M
机构信息
Laboratoire de Microbiologie Appliquée, Centre d'Etudes du Bouchet, Vert-Le-Petit, France.
出版信息
FEMS Microbiol Lett. 1996 Nov 15;145(1):9-16. doi: 10.1111/j.1574-6968.1996.tb08548.x.
Bacillus anthracis can be identified on the basis of the detection of virulence factor genes located on two plasmids, pXO1 and pXO2. Thus isolates lacking both pXO1 and pXO2 are indistinguishable from closely related B. cereus group bacteria. We developed a multiplex PCR assay for characterization of B. anthracis isolates, and simultaneous confirmation of the species identity independent of plasmid content. The assay amplifies lef, cya, pag (pXO1) and cap (pXO2) genes, and a B. anthracis specific chromosomal marker, giving an easy-to-read profile. This system unambiguously identified virulent (pXO1+/2+) and avirulent (pXO1+/2-, pXO1-/2+ and pXO1-/2-) strains of B. anthracis and distinguished "anthrax-like' strains from other B. cereus group bacteria.
炭疽芽孢杆菌可根据位于两个质粒pXO1和pXO2上的毒力因子基因的检测来鉴定。因此，同时缺失pXO1和pXO2的分离株与密切相关的蜡样芽孢杆菌属细菌无法区分。我们开发了一种多重PCR检测方法，用于炭疽芽孢杆菌分离株的鉴定，并能独立于质粒含量同时确认物种身份。该检测方法可扩增lef、cya、pag（pXO1）和cap（pXO2）基因以及炭疽芽孢杆菌特异性染色体标记，生成易于解读的图谱。该系统能够明确鉴定炭疽芽孢杆菌的有毒力菌株（pXO1+/2+）和无毒力菌株（pXO1+/2-、pXO1-/2+和pXO1-/2-），并将“类炭疽”菌株与其他蜡样芽孢杆菌属细菌区分开来。
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