Rapid detection of Bacillus anthracis bloodstream infections by use of a novel assay in the GeneXpert system

Padmapriya P. Banada, Srinidhi Deshpande, Riccardo Russo, Eric Singleton, Darshini Shah, Bhavana Patel, Michele Burday, Ranie Koshy, Qing Wang, Martin Jones, Alexander Gall, Sergey Lokhov, Robert Kwiatkowski, David Persing, Nancy Connell, David Alland

Research output: Contribution to journalArticle

Abstract

Bacillus anthracis 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 B. anthracis 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 B. anthracis DNA into individual PCR mixtures and B. anthracis CFU into human blood. One-milliliter 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-B. anthracis bacteremia or nonbacteremic controls. The assay LODs were 5 genome equivalents per reaction and 10 CFU/ml blood for both the B. anthracis Sterne and V1B strains. There was a 6-log10 dynamic range. Assay specificity was 100% for tests of non-B. anthracis bacterial isolates and patient blood samples. Assay time was less than 90 min. This automated system suitable for point-ofcare detection rapidly identifies B. anthracis directly from blood with high sensitivity. This assay might lead to early detection and more rapid therapy in the event of a bioterrorism attack.

Original languageEnglish (US)
Pages (from-to)2964-2971
Number of pages8
JournalJournal of clinical microbiology
Volume55
Issue number10
DOIs
StatePublished - Oct 1 2017
Externally publishedYes

Keywords

  • Bacillus anthracis
  • Diagnostic
  • Diagnostics
  • GeneXpert
  • Whole blood

ASJC Scopus subject areas

  • Microbiology (medical)

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  • Cite this

    Banada, P. P., Deshpande, S., Russo, R., Singleton, E., Shah, D., Patel, B., Burday, M., Koshy, R., Wang, Q., Jones, M., Gall, A., Lokhov, S., Kwiatkowski, R., Persing, D., Connell, N., & Alland, D. (2017). Rapid detection of Bacillus anthracis bloodstream infections by use of a novel assay in the GeneXpert system. Journal of clinical microbiology, 55(10), 2964-2971. https://doi.org/10.1128/JCM.00466-17