Felder K M, Hoelzle K, Wittenbrink M M, Zeder M, Ehricht R, Hoelzle L E
Institute of Veterinary Bacteriology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland.
Lett Appl Microbiol. 2009 Sep;49(3):324-31. doi: 10.1111/j.1472-765X.2009.02664.x. Epub 2009 Jun 23.
In order to improve the diagnosis of Bacillus anthracis in environmental samples, we established a DNA microarray based on the ArrayTube technology of Clondiag.
Total DNA of a bacterial colony is randomly biotinylated and hybridized to the array. The probes on the array target the virulence genes, the genomic marker gene rpoB, as well as the selective 16S rDNA sequence regions of B. anthracis, of the Bacillus cereus group and of Bacillus subtilis. Eight B. anthracis reference strains were tested and correctly identified. Among the analysed environmental Bacillus isolates, no virulent B. anthracis strain was detected.
This array clearly differentiates B. anthracis from members of the B. cereus group and other Bacillus species in environmental samples by chromosomal (rpoB) and plasmid markers. Additionally, recognition of B. cereus strains harbouring the toxin genes or atypical B. anthracis strains that have lost the virulence plasmids is feasible.
The array is applicable to the complex diagnostics for B. anthracis detection in environmental samples. Because of low costs, high security and easy handling, the microarray is applicable to routine diagnostics.
为提高环境样本中炭疽芽孢杆菌的诊断水平,我们基于Clondiag公司的ArrayTube技术建立了一种DNA微阵列。
细菌菌落的总DNA被随机生物素化并与微阵列杂交。微阵列上的探针靶向炭疽芽孢杆菌、蜡样芽孢杆菌群和枯草芽孢杆菌的毒力基因、基因组标记基因rpoB以及选择性16S rDNA序列区域。对8株炭疽芽孢杆菌参考菌株进行了测试并正确鉴定。在所分析的环境芽孢杆菌分离株中,未检测到有毒力的炭疽芽孢杆菌菌株。
该微阵列通过染色体(rpoB)和质粒标记物,能在环境样本中清晰区分炭疽芽孢杆菌与蜡样芽孢杆菌群成员及其他芽孢杆菌物种。此外,识别携带毒素基因的蜡样芽孢杆菌菌株或已失去毒力质粒的非典型炭疽芽孢杆菌菌株是可行的。
该微阵列适用于环境样本中炭疽芽孢杆菌检测的复杂诊断。由于成本低、安全性高且操作简便,该微阵列适用于常规诊断。