3 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

You might also like
Discover & Discuss Important Research

Keeping up-to-date with research can feel impossible, with papers being published faster than you'll ever be able to read them. That's where Researcher comes in: we're simplifying discovery and making important discussions happen. With over 19,000 sources, including peer-reviewed journals, preprints, blogs, universities, podcasts and Live events across 10 research areas, you'll never miss what's important to you. It's like social media, but better. Oh, and we should mention - it's free.

  • Download from Google Play
  • Download from App Store
  • Download from AppInChina

Researcher displays publicly available abstracts and doesn’t host any full article content. If the content is open access, we will direct clicks from the abstracts to the publisher website and display the PDF copy on our platform. Clicks to view the full text will be directed to the publisher website, where only users with subscriptions or access through their institution are able to view the full article.