This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow E-mail this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ASM journals
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Grunow, R.
Right arrow Articles by Meyer, H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Grunow, R.
Right arrow Articles by Meyer, H.

 Previous Article  |  Next Article 

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.


This article has been cited by other articles:

  • Porsch-Ozcurumez, M., Kischel, N., Priebe, H., Splettstosser, W., Finke, E.-J., Grunow, R. (2004). Comparison of Enzyme-Linked Immunosorbent Assay, Western Blotting, Microagglutination, Indirect Immunofluorescence Assay, and Flow Cytometry for Serological Diagnosis of Tularemia. CVI 11: 1008-1015 [Abstract] [Full Text]  
  • Stenmark, S., Sjostedt, A. (2004). Transfer of specific antibodies results in increased expression of TNF-{alpha} and IL12 and recruitment of neutrophils to the site of a cutaneous Francisella tularensis infection. J Med Microbiol 53: 501-504 [Abstract] [Full Text]  
  • Golovliov, I., Baranov, V., Krocova, Z., Kovarova, H., Sjostedt, A. (2003). An Attenuated Strain of the Facultative Intracellular Bacterium Francisella tularensis Can Escape the Phagosome of Monocytic Cells. Infect. Immun. 71: 5940-5950 [Abstract] [Full Text]  
  • Peruski, A. H., Peruski, L. F. Jr. (2003). Immunological Methods for Detection and Identification of Infectious Disease and Biological Warfare Agents. CVI 10: 506-513 [Full Text]  
  • Tarnvik, A., Berglund, L. (2003). Tularaemia. Eur Respir J 21: 361-373 [Abstract] [Full Text]  
  • Emanuel, P. A., Bell, R., Dang, J. L., McClanahan, R., David, J. C., Burgess, R. J., Thompson, J., Collins, L., Hadfield, T. (2003). Detection of Francisella tularensis within Infected Mouse Tissues by Using a Hand-Held PCR Thermocycler. J. Clin. Microbiol. 41: 689-693 [Abstract] [Full Text]  
  • Ellis, J., Oyston, P. C. F., Green, M., Titball, R. W. (2002). Tularemia. Clin. Microbiol. Rev. 15: 631-646 [Abstract] [Full Text]  
  • Marik, P. E., Bowles, S. (2002). Analytic Reviews: Management of Patients Exposed to Biological and Chemical Warfare Agents. J Intensive Care Med 17: 147-161 [Abstract]  
  • Farlow, J., Smith, K. L., Wong, J., Abrams, M., Lytle, M., Keim, P. (2001). Francisella tularensis Strain Typing Using Multiple-Locus, Variable-Number Tandem Repeat Analysis. J. Clin. Microbiol. 39: 3186-3192 [Abstract] [Full Text]  
  • Dennis, D. T., Inglesby, T. V., Henderson, D. A., Bartlett, J. G., Ascher, M. S., Eitzen, E., Fine, A. D., Friedlander, A. M., Hauer, J., Layton, M., Lillibridge, S. R., McDade, J. E., Osterholm, M. T., O'Toole, T., Parker, G., Perl, T. M., Russell, P. K., Tonat, K., for the Working Group on Civilian Biodefense, (2001). Tularemia as a Biological Weapon: Medical and Public Health Management. JAMA 285: 2763-2773 [Abstract] [Full Text]  


Copyright © 2010 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»