Orthopoxvirus detection in environmental specimens during suspected bioterror attacks: Inhibitory influences of common household products.

Kurth, A., Achenbach, J., Miller, L., Mackay, I. M., Pauli, G. and Nitsche, A. (2008) Orthopoxvirus detection in environmental specimens during suspected bioterror attacks: Inhibitory influences of common household products.. Applied and Environmental Microbiology, 74 1: 32-37. doi:10.1128/AEM.01501-07

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Author Kurth, A.
Achenbach, J.
Miller, L.
Mackay, I. M.
Pauli, G.
Nitsche, A.
Title Orthopoxvirus detection in environmental specimens during suspected bioterror attacks: Inhibitory influences of common household products.
Journal name Applied and Environmental Microbiology   Check publisher's open access policy
ISSN 0099-2240
Publication date 2008-01-01
Year available 2008
Sub-type Article (original research)
DOI 10.1128/AEM.01501-07
Open Access Status File (Publisher version)
Volume 74
Issue 1
Start page 32
End page 37
Total pages 6
Editor Omston, L. N.
Place of publication The United States of America
Publisher American Society for Microbiology
Language eng
Subject 060502 Infectious Agents
060506 Virology
920109 Infectious Diseases
920203 Diagnostic Methods
C1
Abstract After terrorists attacked the United States in 2001, the appearance of letters and other objects containing powdery substances with unknown potentials for biological threat focused attention on the speed, sensitivity, and reliability of diagnostic methods. This study summarizes the abilities and limitations of real-time PCR, electron microscopy (EM), and virus isolation when used to detect potential bioweapons. In particular, we investigated the inhibitory influences of different common household products present in environmental specimens on PCR yield, EM detection, and virus isolation. We used vaccinia virus as a model for orthopoxviruses by spiking it into specimens. In the second part of the study, we describe modifications of diagnostic methods to overcome inhibitory effects. A variety of PCR amplification enhancers, DNA extraction protocols, and applications of internal controls were evaluated to improve diagnostic simplicity, speed, and reliability. As a result, we strongly recommend using at least two different frontline techniques in parallel, e.g., EM and PCR. A positive result obtained by any one of these techniques should be followed by a biological method to confirm the putative diagnosis. Confirmatory methods include virus isolation followed by an agent-specific immunofluorescence assay to confirm the presence of replication-competent particles.
Keyword bioweapons
diagnostics
vaccinia virus
PCR
virus culture
Orthopoxvirus
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: 2009 Higher Education Research Data Collection
 
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Created: Tue, 24 Mar 2009, 00:04:42 EST by Lesley Arnicar on behalf of Clinical Medical Virology Centre