A Standard Method To Inactivate Bacillus anthracis Spores to Sterility via Gamma Irradiation
Abstract
In 2015, a laboratory of the United States Department of Defense (DoD) inadvertently shipped preparations of gamma-irradiated spores of Bacillus anthracis that contained live spores. In response, a systematic evidence-based method for preparing, concentrating, irradiating, and verifying the inactivation of spore materials was developed. We demonstrate the consistency of spore preparations across multiple biological replicates and show that two different DoD institutions independently obtained comparable dose-inactivation curves for a monodisperse suspension of B. anthracis spores containing 3 × 10 10 CFU. Spore preparations from three different institutions and three strain backgrounds yielded similar decimal reduction (D 10 ) values and irradiation doses required to ensure sterility (D SAL ) to the point at which the probability of detecting a viable spore is 10 −6 . Furthermore, spores of a genetically tagged strain of B. anthracis strain Sterne were used to show that high densities of dead spores suppress the recovery of viable spores. Together, we present an integrated method for preparing, irradiating, and verifying the inactivation of spores of B. anthracis for use as standard reagents for testing and evaluating detection and diagnostic devices and techniques.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Jun 15, 2018
- Source ID
- 10.1128/aem.00106-18
Entities
People
- Alena M. Calm
- Alice A. Young
- Amber M. Prugh
- April A. Shea
- Augustus W. Fountain Iii
- Avery V. Quirk
- Bryan A. Rivers
- Casey B. Bernhards
- Christopher K Cote
- Christopher P. Klimko
- David P. Fetterer
- Derrell Mcpherson
- Henry S Gibbons
- Jennifer L. Shoe
- Jeremy A. Miller
- Jerry W. Pfarr
- Joel Bozue
- Josephine S. Esteban-trexler
- Matthew D. Bohmke
- Melissa Hunter
- Neil Kennihan
- Sarah E. Katoski
- Susan L. Welkos
- Todd M. Sickler
- Tony Buhr
- Zachary A. Minter
Organizations
- Defense Threat Reduction Agency
- Edgewood Chemical Biological Center
- Naval Surface Warfare Center