AEM Accepts, published online ahead of print on 6 April 2007

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Appl. Environ. Microbiol. doi:10.1128/AEM.01825-06
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Evaluation of two surface sampling methods for the detection of Erwinia herbicola on a variety of materials using culture and quantitative PCR

Mark P. Buttner^*, Patricia Cruz, Linda D. Stetzenbach, and Tracy Cronin

Harry Reid Center for Environmental Studies, University of Nevada, Las Vegas, 4505 S. Maryland Parkway, Las Vegas, NV 89154-4009; Department of Environmental and Occupational Health, School of Public Health, University of Nevada, Las Vegas, 4505 S. Maryland Parkway, Las Vegas, NV 89154-3063; Technical Support Working Group, Arlington, VA

^* To whom correspondence should be addressed. Email: buttner{at}unlv.nevada.edu.

This research was designed to evaluate surface sampling protocols for use with culture and quantitative polymerase chain reaction (QPCR) amplification assay for the detection of the gram-negative bacterial biothreat simulant, Erwinia herbicola, on a variety of surface materials. Surfaces selected for evaluation were wood laminate, glass/computer monitor screens, metal file cabinet, plastic arena seats, nylon seat cushions, finished concrete flooring, and vinyl tile flooring. Laboratory and test chamber studies were performed to evaluate two sampling methods, a sponge and a macrofoam swab, for the detection of E. herbicola on surface materials. In laboratory trials, seven materials were inoculated with a known concentration of E. herbicola cells, and samples were collected from the surfaces of the materials to determine sampling efficiencies. Culture analysis was ineffective for assessing collection efficiency of E. herbicola because very few culturable cells were obtained from surface samples. QPCR demonstrated that E. herbicola DNA was present in high concentrations in all surface samples and sampling efficiencies ranged from 0.7% to 52.2%, depending on the sampling method and the surface material. The swab was generally more efficient than the sponge for collection of E. herbicola from surfaces. Test chamber trials were also performed in which E. herbicola was aerosolized into the chamber and allowed to settle onto test materials. Surface sampling results supported those obtained in laboratory trials. The results of this study demonstrate the capabilities of QPCR to enhance detection and enumeration of biocontaminants on surface materials and provide information on the comparability of sampling methods.