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Steinmetz, I; Rheinheimer, C; Hübner, I; Bitter-Suermann, D.
Genus-specific epitope on the 60-kilodalton Legionella heat shock protein recognized by a monoclonal antibody.
J Clin Microbiol. 1991; 29(2):346-354 Doi: 10.1128/JCM.29.2.346-354.1991 [OPEN ACCESS]
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Leading authors Med Uni Graz: Steinmetz Ivo
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Abstract:: A monoclonal antibody (MAb) immunoglobulin G2a (2125) was produced against a 60-kDa Legionella heat shock protein (HSP), recognizing a unique epitope common to all species of the genus Legionella. The antibody reacted in the immunoblot with 59 Legionella species and serogroups that were tested and showed no cross-reactivity with other bacteria, including Acinetobacter spp., Bordetella spp., Pseudomonas spp., Mycobacterium spp., and Escherichia coli. Two other MAbs (2122 and 2130) reacted with the 60-kDa Legionella protein as well but showed different cross-reactivities with other gram-negative bacteria in the same molecular mass range. The genus-specific MAb 2125 as well as the cross-reacting MAbs 2122 and 2130 were shown to be reactive with the expressed protein of the cloned gene of the 60-kDa HSP of Legionella micdadei and Legionella pneumophila. These antibodies demonstrate that Legionella-specific and nonspecific epitopes are present on this protein. A sandwich enzyme-linked immunosorbent assay (ELISA) in which the genus-specific MAb is used both as a capture antibody and as a biotinylated second antibody has been established. With this test it is possible to detect Legionella whole cells, sonicated cells, and cell fractions containing the 60-kDa HSP. The main part of the 60-kDa HSP is found in the cytoplasmic fraction. The sandwich ELISA can be used to demonstrate the increased expression of the 60-kDa protein in Legionella cells following heat shock as well as marked differences in the detection of the 60-kDa HSP on whole cells of different Legionella strains. The high specificity and sensitivity of the sandwich ELISA for sonicated cells might be very useful to screen on a genus level for Legionella cells or the 60-kDa antigen in environmental isolates or body fluids of patients.
Find related publications in this database (using NLM MeSH Indexing): Antibodies, Bacterial -; Antibodies, Monoclonal -; Antibody Specificity -; Antigens, Bacterial -; Bacterial Proteins - chemistry; Bacterial Proteins - immunology; Cross Reactions -; Enzyme-Linked Immunosorbent Assay -; Epitopes -; Heat-Shock Proteins - chemistry; Heat-Shock Proteins - immunology; Humans -; Legionella - immunology; Legionella - isolation & purification; Legionellosis - diagnosis; Molecular Weight -; Species Specificity -