Open AccessProduction of a monoclonal antibody specific for a Flavobacterium species isolated from soil
Author(s) -
Mason Julie,
Burns Richard G.
Publication year - 1990
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1990.tb03953.x
Subject(s) - flavobacterium , immunogold labelling , monoclonal antibody , biology , bacteria , microbiology and biotechnology , immunofluorescence , antigen , gram negative bacteria , pseudomonas , antibody , biochemistry , escherichia coli , immunology , genetics , gene
Hybridomas secreting monoclonal antibodies (MABs) specific for a soil Flavobacterium species (P25) were isolated. The MAB (D10) was used to target P25 using an enzyme‐linked immunosorbant assay (ELISA) and indirect immunofluorescence. Cross‐reactivity of the MAB with other Gram‐negative bacteria (including Flavobacterium spp.) and a number of Gram‐positive bacteria was investigated but none were found. Cross‐reactivity with other orange/yellow pigmented Gram‐negative rods ( Pseudomonas/Flavobacterium type) isolated from the soil into which P25 has been introduced in field experiments was also assessed using a modified colony blotting procedure. None of the indigenous species tested were recognised by the monoclonal antibody, thereby allowing unambiguous identification of P25 in soil. The MAB D10 was shown to recognise P25 growth under low‐nutrient or stored under starvation conditions, suggesting that the antigen is a constitutive component of the cell and that the microorganism should be detected in oligotrophic environments such as soil. The pattern of fluorescence of P25 gave a clear indication of the localisation of the antigen in the outer membrane/cell wall region, and this was confirmed by immunogold labelling. Preliminary studies on the limits of detection of P25 using immunofluorescence suggest that densities as low as 20 bacteria g −1 soil can be enumerated.