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Artificial plasmid engineered to simulate multiple biological threat agents.

Abstract：

: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.

journal_name

Appl Microbiol Biotechnol

journal_title

Applied microbiology and biotechnology

authors

Carrera M,Sagripanti JL

doi

10.1007/s00253-008-1715-8

subject

Has Abstract

pub_date

2009-01-01 00:00:00

pages

1129-39

issue

6

eissn

0175-7598

issn

1432-0614

journal_volume

81

pub_type

杂志文章

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