Real-time PCR system targeting a chromosomal marker specific for Bacillus anthracis

被引:57
作者
Antwerpen, Markus H. [1 ]
Zimmermann, Pia [1 ]
Bewley, Kevin [2 ]
Frangoulidis, Dimitrios [1 ]
Meyer, Hermann [1 ]
机构
[1] Bundeswehr Inst Microbiol, D-80937 Munich, Germany
[2] Hlth Protect Agcy, Salisbury, Wilts, England
关键词
Bacillus anthracis; Real-time PCR; Chromosomal marker; dhp61; BA5345;
D O I
10.1016/j.mcp.2008.06.001
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Specific identification of Bacillus anthracis and differentiation from closely related Bacillus cereus and Bacillus thuringiensis strains is still a major diagnostic problem. Commercially available diagnostic kits targeting plasmid-markers cannot differentiate between B. anthracis, non-anthracis Bacillus species harbouring anthrax-specific virulence plasmids, and plasmidless B. anthracis strains. A TaqMan (R) PCR assay was designed targeting sequences of gene locus BA_5345 of the B. anthracis strain Ames. Specificity was determined by using a panel of 328 Bacillus strains; sensitivity was determined by probit analysis. All B. anthracis isolates (n = 92) were specifically detected by using the genomic TaqMan (R) PCR assay whereas 236 strains belonging to 19 Bacillus species other than B. anthracis were PCR negative. The detection limit was determined to be 12.7 copies per reaction (95% confidence interval 10.2-17.5 copies). Here we present an extensively evaluated and - to our current knowledge - specific TaqMan (R) PCR assay for the detection of B. anthracis based on a chromosomal marker. (C) 2008 Elsevier Ltd. All rights reserved.
引用
收藏
页码:313 / 315
页数:3
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