Journal of Clinical Microbiology
Journal of Clinical Microbiology
5 years ago

Rapid Detection of Bacillus anthracis Blood Stream Infections Using a Novel Assay in the GeneXpert System.

Singleton E, Alland D, Connell N, Kwiatkowski R, Koshy R, Deshpande S, Wang Q, Russo R, Jones M, Persing D, Gall A, Lokhov S, Burday M, Patel B, Banada PP, Shah D
BACKGROUND.Bacillus anthracis is a tier1 select agent with the potential to quickly cause severe disease. Rapid identification of this pathogen may accelerate treatment and reduce mortality in the event of a bioterrorism attack. We developed a rapid and sensitive assay to detect B. anthracis bacteremia using a system that is suitable for point of care testing.METHODS. A filter-based cartridge that included both sample processing and PCR amplification functions was loaded with all reagents needed for sample processing and multiplex nested PCR. Assay limit of detection (LOD), and dynamic range were determined by spiking B. anthracis DNA into individual PCR reactions, and B. anthracis colony forming units (CFU) into human blood. One-ml blood samples were added to the filter-based detection cartridge and tested for B. anthracis on a GeneXpert instrument. Assay specificity was determined by testing blood spiked with non-anthrax bacterial isolates, or by testing blood samples drawn from patients with concurrent non-anthracis bacteremia or non-bacteremic controls.RESULTS. The assay LOD was 5 genome equivalents per reaction, and 10 CFU/ml blood for both B. anthracis Sterne and V1B strains. There was a six log10 dynamic range. Assay specificity was 100% for tests of non-anthracis bacterial isolates and patient blood samples. Assay time was less than 90 minutes.CONCLUSIONS. This automated system suitable for point of care detection rapidly identifies B. anthracis directly from blood with high sensitivity. This assay could lead to early detection and more rapid therapy in the event of a bioterrorism attack.

Publisher URL: https://www.ncbi.nlm.nih.gov/pubmed/28747367

DOI: PubMed:28747367

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