This Article Full Text Full Text (PDF) Other Versions of this Article: AEM.01825-06v1 73/11/3505    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Buttner, M. P. Articles by Cronin, T. Search for Related Content PubMed PubMed Citation Articles by Buttner, M. P. Articles by Cronin, T. Agricola Articles by Buttner, M. P. Articles by Cronin, T.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, June 2007, p. 3505-3510, Vol. 73, No. 11
0099-2240/07/$08.00+0     doi:10.1128/AEM.01825-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Evaluation of Two Surface Sampling Methods for Detection of Erwinia herbicola on a Variety of Materials by Culture and Quantitative PCR

Mark P. Buttner,^1* Patricia Cruz,¹ Linda D. Stetzenbach,² and Tracy Cronin^3,

Harry Reid Center for Environmental Studies, University of Nevada, Las Vegas, 4505 S. Maryland Parkway, Las Vegas, Nevada 89154-4009,¹ Department of Environmental and Occupational Health, School of Public Health, University of Nevada, Las Vegas, 4505 S. Maryland Parkway, Las Vegas, Nevada 89154-3063,² Technical Support Working Group, Arlington, Virginia³

Received 2 August 2006/ Accepted 29 March 2007

This research was designed to evaluate surface sampling protocols for use with culture and quantitative PCR (QPCR) amplification assay for detection of the gram-negative bacterial biothreat simulant Erwinia herbicola on a variety of surface materials. Surfaces selected for evaluation were wood laminate, glass and computer monitor screens, metal file cabinets, 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 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 E. herbicola collection efficiency because very few culturable cells were obtained from surface samples. QPCR demonstrated that E. herbicola DNA was present in high concentrations on all of the 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 the detection and enumeration of biocontaminants on surface materials and provide information on the comparability of sampling methods.

Published ahead of print on 6 April 2007.

Present address: TD Cronin & Associates, LLC, Alexandria, VA.