| Optimal swab processing recovery method for detection of bioterrorism-related Francisella tularensis by real-time PCR. | |
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MedLine Citation:
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PMID: 20654658 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Francisella tularensis, the etiological agent of tularemia, is regarded as a potential bioterrorism agent. The advent of bioterrorism has heightened awareness of the need for validated methods for processing environmental samples. In this study we determined the optimal method for processing environmental swabs for the recovery and subsequent detection of F. tularensis by the use of real-time PCR assays. Four swab processing recovery methods were compared: heat, sonication, vortexing, and the Swab Extraction Tube System (SETS). These methods were evaluated using cotton, foam, polyester and rayon swabs spiked with six pathogenic strains of F. tularensis. Real-time PCR analysis using a multi-target 5'nuclease assay for F. tularensis showed that the use of the SETS method resulted in the best limit of detection when evaluated using multiple strains of F. tularensis. We demonstrated also that the efficiency of F. tularensis recovery from swab specimens was not equivalent for all swab processing methodologies and, thus, that this variable can affect real-time PCR assay sensitivity. The effectiveness of the SETS method was independent of the automated DNA extraction method and real-time PCR platforms used. In conclusion, diagnostic laboratories can now potentially incorporate the SETS method into specimen processing protocols for the rapid and efficient detection of F. tularensis by real-time PCR during laboratory bioterrorism-related investigations. |
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Authors:
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Roblena E Walker; Jeannine M Petersen; Kenyatta W Stephens; Leslie A Dauphin |
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Publication Detail:
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Type: Evaluation Studies; Journal Article Date: 2010-07-21 |
Journal Detail:
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Title: Journal of microbiological methods Volume: 83 ISSN: 1872-8359 ISO Abbreviation: J. Microbiol. Methods Publication Date: 2010 Oct |
Date Detail:
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Created Date: 2010-09-10 Completed Date: 2010-12-13 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8306883 Medline TA: J Microbiol Methods Country: Netherlands |
Other Details:
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Languages: eng Pagination: 42-7 Citation Subset: IM |
Copyright Information:
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Published by Elsevier B.V. |
Affiliation:
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Bioterrorism Rapid Response and Advanced Technology (BRRAT) Laboratory, Division of Bioterrorism Preparedness and Response (DBPR), Centers for Disease Control and Prevention (CDC), Atlanta, GA, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Bioterrorism* DNA, Bacterial / genetics, isolation & purification Francisella tularensis / genetics, isolation & purification* Humans Polymerase Chain Reaction / methods* Tularemia / microbiology |
| Chemical | |
Reg. No./Substance:
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0/DNA, Bacterial |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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