Sequencing of 16S rRNA gene: a rapid tool for identification of Bacillus anthracis

• Published in: Emerging infectious diseases 2002-10, Vol.8 (10), p.1117-1123
• Main Authors: Sacchi, Claudio T, Whitney, Anne M, Mayer, Leonard W, Morey, Roger, Steigerwalt, Arnold, Boras, Arijana, Weyant, Robin S, Popovic, Tanja
• Format: Article
• Language: English
• Subjects: 16S rRNA gene; Analysis; Anthrax; Anthrax - diagnosis; Anthrax - microbiology; Bacillus anthracis; Bacillus anthracis - genetics; Bacillus anthracis - isolation & purification; Bacillus cereus - genetics; Bacillus cereus - isolation & purification; Bacillus thuringiensis - genetics; Bacillus thuringiensis - isolation & purification; Bioterrorism; Dispatch; Genes, Bacterial - genetics; Genetic aspects; Genetic research; Genetic Variation - genetics; Humans; Molecular Sequence Data; Polymerase Chain Reaction; RNA, Bacterial - genetics; RNA, Ribosomal, 16S - genetics; Sequence Analysis, DNA - methods; United States
• ISSN: 1080-6040; 1080-6059
• DOI: 10.3201/eid0810.020391

In a bioterrorism event, a tool is needed to rapidly differentiate Bacillus anthracis from other closely related spore-forming Bacillus species. During the recent outbreak of bioterrorism-associated anthrax, we sequenced the 16S rRNA generom these species to evaluate the potential of 16S rRNA gene sequencing as a diagnostic tool. We found eight distinct 16S types among all 107 16S rRNA gene seqs fuences that differed from each other at 1 to 8 positions (0.06% to 0.5%). All 86 B. anthracis had an identical 16S gene sequence, designated type 6; 16S type 10 was seen in all B. thuringiensis strains; six other 16S types were found among the 10 B. cereus strains. This report describes the first demonstration of an exclusive association of a distinct 16S sequence with B. anthracis. Consequently, we were able to rapidly identify suspected isolates and to detect the B. anthracis 16S rRNA gene directly from culture-negative clinical specimens from seven patients with laboratory-confirmed anthrax.