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Clinical and Diagnostic Laboratory Immunology, January 2000, p. 86-90, Vol. 7, No. 1
1071-412X/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Detection of Francisella tularensis in Biological Specimens Using a Capture Enzyme-Linked Immunosorbent Assay, an Immunochromatographic Handheld Assay, and a PCR

Roland Grunow,1,* Wolf Splettstoesser,1 Sahra McDonald,2 Christian Otterbein,1 Tom O'Brien,3 Cecilia Morgan,3 Jennifer Aldrich,3 Erwin Hofer,4 Ernst-Jürgen Finke,1 and Hermann Meyer1

Institute of Microbiology, Federal Armed Forces Medical Academy, 80937 Munich, Germany1; Porton Down, Salisbury, United Kingdom2; Naval Medical Research Institute, Bethesda, Maryland3; and Federal Institute for Control of Infectious Diseases of Animals, 2340 Mödling, Austria4

Received 30 March 1999/Returned for modification 19 August 1999/Accepted 20 October 1999

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 103 to 104 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.

* Corresponding author. Mailing address: Institute of Microbiology, Federal Armed Forces Medical Academy, Neuherbergstr. 11, D-80937 Munich, Germany. Phone: 41-89-3168 3277. Fax: 49-89-3168 3292. E-mail: tbl01cn{at}mail.lrz-muenchen.de.

Clinical and Diagnostic Laboratory Immunology, January 2000, p. 86-90, Vol. 7, No. 1
1071-412X/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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