Affiliation:
1. Institute of Microbiology, Federal Armed Forces Medical Academy, 80937 Munich, Germany1;
2. Porton Down, Salisbury, United Kingdom2;
3. Naval Medical Research Institute, Bethesda, Maryland3; and
4. Federal Institute for Control of Infectious Diseases of Animals, 2340 Mödling, Austria4
Abstract
ABSTRACT
The early detection of
Francisella tularensis
, the causative agent of tularemia, is important for adequate treatment by antibiotics and the outcome of the disease. Here we describe a new capture enzyme-linked immunosorbent assay (cELISA) based on monoclonal antibodies specific for lipopolysaccharide (LPS) of
Francisella tularensis
subsp.
holarctica
and
Francisella tularensis
subsp.
tularensis
. No cross-reactivity with
Francisella tularensis
subsp.
novicida
,
Francisella philomiragia
, and a panel of other possibly related bacteria, including
Brucella
spp.,
Yersinia
spp.,
Escherichia coli
, and
Burkholderia
spp., was observed. The detection limit of the assay was 10
3
to 10
4
bacteria/ml. This sensitivity was achieved by solubilization of the LPS prior to the cELISA. In addition, a novel immunochromatographic membrane-based handheld assay (HHA) and a PCR, targeting sequences of the 17-kDa protein (TUL4) gene of
F. tularensis
, were used in this study. Compared to the cELISA, the sensitivity of the HHA was about 100 times lower and that of the PCR was about 10 times higher. All three techniques were successfully applied to detect
F. tularensis
in tissue samples of European brown hares (
Lepus europaeus
). Whereas all infected samples were recognized by the cELISA, those with relatively low bacterial load were partially or not detected by PCR and HHA, probably due to inhibitors or lack of sensitivity. In conclusion, the HHA can be used as a very fast and simple approach to perform field diagnosis to obtain a first hint of an infection with
F. tularensis
, especially in emergent situations. In any suspect case, the diagnosis should be confirmed by more sensitive techniques, such as the cELISA and PCR.
Publisher
American Society for Microbiology
Subject
Microbiology (medical),Clinical Biochemistry,Immunology,Immunology and Allergy
Cited by
130 articles.
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