Rapid Detection of Bacillus anthracis Bloodstream Infections by Use of a Novel Assay in the GeneXpert System

• Published in: Journal of clinical microbiology 2017-10, Vol.55 (10), p.2964-2971
• Main Authors: Banada, Padmapriya P, Deshpande, Srinidhi, Russo, Riccardo, Singleton, Eric, Shah, Darshini, Patel, Bhavana, Burday, Michele, Koshy, Ranie, Wang, Qing, Jones, Martin, Gall, Alexander, Lokhov, Sergey, Kwiatkowski, Robert, Persing, David, Connell, Nancy, Alland, David
• Format: Article
• Language: English
• Subjects: Bacteriology
• ISSN: 0095-1137; 1098-660X
• DOI: 10.1128/JCM.00466-17

is a tier 1 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 bacteremia using a system that is suitable for point-of-care testing. 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. The assay limit of detection (LOD) and dynamic range were determined by spiking DNA into individual PCR mixtures and CFU into human blood. One-milliliter blood samples were added to the filter-based detection cartridge and tested for 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- bacteremia or nonbacteremic controls. The assay LODs were 5 genome equivalents per reaction and 10 CFU/ml blood for both the Sterne and V1B strains. There was a 6-log dynamic range. Assay specificity was 100% for tests of non- bacterial isolates and patient blood samples. Assay time was less than 90 min. This automated system suitable for point-of-care detection rapidly identifies directly from blood with high sensitivity. This assay might lead to early detection and more rapid therapy in the event of a bioterrorism attack.