Document Detail

Artificial plasmid engineered to simulate multiple biological threat agents.
MedLine Citation:
PMID:  18923830     Owner:  NLM     Status:  MEDLINE    
The objective of this study was to develop a non-virulent simulant to replace several virulent organisms during the development of detection and identification methods for biological threat agents. We identified and selected specific genes to detect Yersinia pestis, Francisella tularensis, Burkholderia mallei, Burkholderia pseudomallei, Rickettsia sp., Coxiella burnetii, Brucella sp., enterohemorrhagic Escherichia coli O157:H7, Bacillus anthracis, and variola (smallpox) virus. We then designed and engineered a non-infectious simulant that included the nucleic-acid signature of each microorganism in a single chimerical molecule. Here, we reported an approach that by direct (de novo) chemical synthesis permitted the production of a single chimerical construct 2,040 bp long that included the nucleic-acid signature of the bacterial and viral biological threat agents listed above without requiring access to these agents. Sequences corresponding to each one of the biological agents in the synthetic simulant were amplified by PCR, resulting in amplicons of the expected length, of similar intensity, and without any detectable unspecific products. The novel simulant described here could reduce the need for infectious agents in the development of detection and diagnostic methods and should also be useful as a non-virulent positive control in nucleic-acid-based tests against biological threat agents.
Monica Carrera; Jose-Luis Sagripanti
Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2008-10-16
Journal Detail:
Title:  Applied microbiology and biotechnology     Volume:  81     ISSN:  1432-0614     ISO Abbreviation:  Appl. Microbiol. Biotechnol.     Publication Date:  2009 Jan 
Date Detail:
Created Date:  2008-12-24     Completed Date:  2009-01-23     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8406612     Medline TA:  Appl Microbiol Biotechnol     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  1129-39     Citation Subset:  IM    
Edgewood Chemical Biological Center, Research Development and Engineering Command US Army, AMSRD-ECB-RT, Aberdeen Proving Ground, Aberdeen, MD 21010-5424, USA.
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MeSH Terms
Bacteria / genetics*,  isolation & purification
Biological Warfare Agents*
DNA, Recombinant*
Genes, Bacterial
Genes, Viral
Viruses / genetics*,  isolation & purification
Reg. No./Substance:
0/DNA, Recombinant

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