title: Real-Time PCR Identification of Unique Bacillus anthracis Sequences.
abstract: Bacillus anthracis is a spore-forming, Gram-positive microorganism. It is a causative agent of anthrax, a highly infectious disease. It belongs to the "Bacillus cereus group", which contains other closely related species, including Bacillus cereus, Bacillus thuringiensis, Bacillus mycoides, Bacillus weihenstephanensis, and Bacillus pseudomycoides. B. anthracis naturally occurs in soil environments. The BA5345 genetic marker was used for highly specific detection of B. anthracis with TaqMan probes. The detection limit of a real-time PCR assay was estimated at the level of 16.9 copies (CI95% - 37.4 to 37.86, SD = 0.2; SE = 0.118). Oligonucleotides designed for the targeted sequences (within the tested locus) revealed 100 % homology to B. anthracis strain reference sequences deposited in the database (NCBI) and high specificity to all tested B. anthracis strains. Additional in silico analysis of plasmid markers pag and cap genes with B. anthracis strains included in the database was carried out. Our study clearly indicates that the BA5345 marker can be used with success as a chromosomal marker in routine identification of B. anthracis; moreover, detection of plasmid markers indicates virulence of the examined strains.
authors.affiliation: Biological Threats Identification and Countermeasure Centre of the General Karol Kaczkowski Military Institute of Hygiene and Epidemiology, Puławy, Poland.
authors.forename: P
authors.initials: P
authors.lastname: Cieślik
authors.affiliation: Warsaw Medical University, Second Faculty of Medicine, Department of Epidemiology, Warsaw, Poland.
authors.forename: J
authors.initials: J
authors.lastname: Knap
authors.affiliation: Biological Threats Identification and Countermeasure Centre of the General Karol Kaczkowski Military Institute of Hygiene and Epidemiology, Puławy, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Kolodziej
authors.affiliation: Biological Threats Identification and Countermeasure Centre of the General Karol Kaczkowski Military Institute of Hygiene and Epidemiology, Puławy, Poland.
authors.forename: T
authors.initials: T
authors.lastname: Mirski
authors.affiliation: Biological Threats Identification and Countermeasure Centre of the General Karol Kaczkowski Military Institute of Hygiene and Epidemiology, Puławy, Poland.
authors.forename: J
authors.initials: J
authors.lastname: Joniec
authors.affiliation: Biological Threats Identification and Countermeasure Centre of the General Karol Kaczkowski Military Institute of Hygiene and Epidemiology, Puławy, Poland.
authors.forename: G
authors.initials: G
authors.lastname: Graniak
authors.affiliation: Biological Threats Identification and Countermeasure Centre of the General Karol Kaczkowski Military Institute of Hygiene and Epidemiology, Puławy, Poland.
authors.forename: D
authors.initials: D
authors.lastname: Zakowska
authors.affiliation: General Karol Kaczkowski Military Institute of Hygiene and Epidemiology, Epidemiology Department, Warsaw, Poland.
authors.forename: I
authors.initials: I
authors.lastname: Winnicka
authors.affiliation: Biological Threats Identification and Countermeasure Centre of the General Karol Kaczkowski Military Institute of Hygiene and Epidemiology, Puławy, Poland.
authors.forename: A
authors.initials: A
authors.lastname: Bielawska-Drózd
mesh_terms: D000942:Antigens, Bacterial / Q000235:genetics; D001408:Bacillus anthracis / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D001409:Bacillus cereus / Q000235:genetics / Q000472:pathogenicity; D001426:Bacterial Proteins / Q000235:genetics*; D001427:Bacterial Toxins / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis; D005819:Genetic Markers; D010957:Plasmids / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D037521:Virulence Factors / Q000235:genetics
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004269:DNA, Bacterial; D005819:Genetic Markers; D037521:Virulence Factors; C030325:anthrax toxin