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Applied and Environmental Microbiology, July 2009, p. 4297-4306, Vol. 75, No. 13
0099-2240/09/$08.00+0     doi:10.1128/AEM.02549-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Recovery Efficiency and Limit of Detection of Aerosolized Bacillus anthracis Sterne from Environmental Surface Samples {triangledown}

Cheryl Fairfield Estill,1* Paul A. Baron,1 Jeremy K. Beard,2 Misty J. Hein,1 Lloyd D. Larsen,2 Laura Rose,3 Frank W. Schaefer III,4 Judith Noble-Wang,3 Lisa Hodges,3 H. D. Alan Lindquist,4 Gregory J. Deye,1 and Matthew J. Arduino3

Centers for Disease Control and Prevention, National Institute for Occupational Safety and Health, 4676 Columbia Parkway, Cincinnati, Ohio 45226,1 Dugway Proving Ground, P.O. Box 217, Dugway, Utah 84022,2 Centers for Disease Control and Prevention, Center for Infectious Diseases, 1600 Clifton Road, N.E., Atlanta, Georgia 30333,3 U.S. Environmental Protection Agency, National Homeland Security Research Center, 26 W. M. L. King Drive, Cincinnati, Ohio 452684

Received 7 November 2008/ Accepted 2 May 2009

After the 2001 anthrax incidents, surface sampling techniques for biological agents were found to be inadequately validated, especially at low surface loadings. We aerosolized Bacillus anthracis Sterne spores within a chamber to achieve very low surface loading (ca. 3, 30, and 200 CFU per 100 cm2). Steel and carpet coupons seeded in the chamber were sampled with swab (103 cm2) or wipe or vacuum (929 cm2) surface sampling methods and analyzed at three laboratories. Agar settle plates (60 cm2) were the reference for determining recovery efficiency (RE). The minimum estimated surface concentrations to achieve a 95% response rate based on probit regression were 190, 15, and 44 CFU/100 cm2 for sampling steel surfaces and 40, 9.2, and 28 CFU/100 cm2 for sampling carpet surfaces with swab, wipe, and vacuum methods, respectively; however, these results should be cautiously interpreted because of high observed variability. Mean REs at the highest surface loading were 5.0%, 18%, and 3.7% on steel and 12%, 23%, and 4.7% on carpet for the swab, wipe, and vacuum methods, respectively. Precision (coefficient of variation) was poor at the lower surface concentrations but improved with increasing surface concentration. The best precision was obtained with wipe samples on carpet, achieving 38% at the highest surface concentration. The wipe sampling method detected B. anthracis at lower estimated surface concentrations and had higher RE and better precision than the other methods. These results may guide investigators to more meaningfully conduct environmental sampling, quantify contamination levels, and conduct risk assessment for humans.

* Corresponding author. Mailing address: Centers for Disease Control and Prevention, National Institute for Occupational Safety and Health, MS R-14, 4676 Columbia Parkway, Cincinnati, OH 45226. Phone: (513) 841-4476. Fax: (513) 841-4486. E-mail: CEstill{at}cdc.gov

{triangledown} Published ahead of print on 8 May 2009.

Applied and Environmental Microbiology, July 2009, p. 4297-4306, Vol. 75, No. 13
0099-2240/09/$08.00+0     doi:10.1128/AEM.02549-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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