Affiliation:
1. Bacterial Zoonoses Branch, Division of Vector-Borne Infectious Diseases, National Center for Infectious Disease, Centers for Disease Control and Prevention, Ft. Collins, Colorado 80522
Abstract
ABSTRACT
Tularemia is the zoonotic disease caused by the gram-negative coccobacillus
Francisella tularensis.
Its wide distribution in the environment poses a challenge for understanding the transmission, ecology, and epidemiology of the disease.
F. tularensis
is also considered a potential biological weapon due to its extreme infectivity. We have developed a multitarget real-time TaqMan PCR assay capable of rapidly and accurately detecting
F. tularensis
in complex specimens. Targeted regions included the IS
Ftu2
element and the
23kDa
,
fopA
, and
tul4
genes. Analysis of the four TaqMan assays demonstrated that three (IS
Ftu2
,
23kDa
, and
tul4
) performed within our established criterion of a detection limit of one organism. The combined use of the three assays was highly specific, displaying no cross-reactivity with the non-
Francisella
bacteria tested and capable of differentially diagnosing both
F. tularensis
and
Francisella philomiragia
. When the multitarget TaqMan assay (IS
Ftu2
,
23kDa
, and
tul4
) was compared to culturing, using environmentally contaminated specimens, the TaqMan PCR assay was significantly more sensitive than culturing (
P
≤ 0.05). The sensitive and specific nature of this rapid multitarget TaqMan assay provides a valuable new tool that with future evaluations can be used for analyzing clinical specimens, field samples during bioterrorism threat assessment, and samples from outbreaks and for improving our understanding of the ecology and environmental prevalence of
F. tularensis
.
Publisher
American Society for Microbiology
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