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Applied and Environmental Microbiology, December 2004, p. 7040-7045, Vol. 70, No. 12
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.12.7040-7045.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Evaluation of the Biological Sampling Kit (BiSKit) for Large-Area Surface Sampling

Mark P. Buttner,1* Patricia Cruz,1 Linda D. Stetzenbach,1 Amy K. Klima-Comba,1 Vanessa L. Stevens,1 and Peter A. Emanuel2

Harry Reid Center for Environmental Studies, University of Nevada, Las Vegas, Las Vegas, Nevada,1 Critical Reagents Program, Chemical Biological Medical Systems, Department of Defense, Aberdeen Proving Ground, Maryland2

Received 10 February 2004/ Accepted 12 July 2004

Current surface sampling methods for microbial contaminants are designed to sample small areas and utilize culture analysis. The total number of microbes recovered is low because a small area is sampled, making detection of a potential pathogen more difficult. Furthermore, sampling of small areas requires a greater number of samples to be collected, which delays the reporting of results, taxes laboratory resources and staffing, and increases analysis costs. A new biological surface sampling method, the Biological Sampling Kit (BiSKit), designed to sample large areas and to be compatible with testing with a variety of technologies, including PCR and immunoassay, was evaluated and compared to other surface sampling strategies. In experimental room trials, wood laminate and metal surfaces were contaminated by aerosolization of Bacillus atrophaeus spores, a simulant for Bacillus anthracis, into the room, followed by settling of the spores onto the test surfaces. The surfaces were sampled with the BiSKit, a cotton-based swab, and a foam-based swab. Samples were analyzed by culturing, quantitative PCR, and immunological assays. The results showed that the large surface area (1 m2) sampled with the BiSKit resulted in concentrations of B. atrophaeus in samples that were up to 10-fold higher than the concentrations obtained with the other methods tested. A comparison of wet and dry sampling with the BiSKit indicated that dry sampling was more efficient (efficiency, 18.4%) than wet sampling (efficiency, 11.3%). The sensitivities of detection of B. atrophaeus on metal surfaces were 42 ± 5.8 CFU/m2 for wet sampling and 100.5 ± 10.2 CFU/m2 for dry sampling. These results demonstrate that the use of a sampling device capable of sampling larger areas results in higher sensitivity than that obtained with currently available methods and has the advantage of sampling larger areas, thus requiring collection of fewer samples per site.

* Corresponding author. Mailing address: Harry Reid Center for Environmental Studies, University of Nevada, Las Vegas, 4505 S. Maryland Parkway, Las Vegas, NV 89154-4009. Phone: (702) 895-1418. Fax: (702) 895-2688. E-mail: buttner{at}unlv.nevada.edu.

Applied and Environmental Microbiology, December 2004, p. 7040-7045, Vol. 70, No. 12
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.12.7040-7045.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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