Method of testing for the presence of Salmonella serotypes expressing Salmonella enteritidis fimbrial antigen (SEFA) and reagents therefore
AssigneeThe Minister of Agriculture, Fisheries and Food in Her Britannic Majesty's Government of the United Kingdom of Great Britian and Northern Ireland
InventorChristopher J. Thorns
Life of this patent
Estimating the term…
Abstract
A method of testing for the presence of Salmonella serotypes S. enteritidis and S. dublin is provided. Novel monoclonal antibodies are used to detect the presence of an epitope specific for these serotypes in cultures which have been grown on selected media which enhance the expression of said epitope in fimbrial sites. Test kits utilizing the antigen or its epitopic parts, antibodies and/or the media are further provided.
Claims31
A method of testing a sample for the presence of microorganisms for Salmonella serotypes expressing Salmonella enteritidis fimbrial antigen (SEFA) comprising the steps of:
- (a) exposing a sample suspected of containing the microorganisms, or SEFA to an antibody which specifically binds to the antigen specifically bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902 or an antibody which specifically binds the epitope bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902;
- (b) detecting antibody-antigen specific binding, wherein antigen-antibody specific binding is indicative of the presence of microorganisms selected from the group consisting of S. enteritidis, S. dublin, S. moscow and S. blegdam, and the absence of antibody-antigen specific binding is indicative of the absence of S. enteritidis.
A method of testing for the presence of a previous or current infection with Salmonella serotypes expressing SEFA comprising the steps of:
- (a) exposing said SEFA to a biological specimen obtained from a subject suspected of having a current or a previous Salmonella infection, wherein said SEFA specifically binds an antibody which specifically binds to the antigen specifically bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902 or an antibody which specifically binds the epitope bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902;
- then
- (b) detecting antibody-antigen specific binding wherein the presence of antibody-antigen specific binding is indicative of the presence of a previous or current Salmonella infection with microorganisms selected from the group consisting of S. enteritidis, S. dublin, S. moscow and S. blegdam, and the absence of antibody-antigen specific binding is indicative of the absence of a previous or current infection with S. enteritidis.
A method of determining whether a Salmonella serotype belongs to either a group consisting of S. enteritidis, S. moscow and S. blegdam or a group consisting of S. dublin, S. moscow and S. blegdam comprising the steps of:
- (a) exposing a sample suspected of containing at least one of said Salmonella serotypes to an antibody which specifically binds to the antigen specifically bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902 or an antibody which specifically binds the epitope bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902 and then detecting antibody-antigen binding, wherein antibody-antigen specific binding is indicative of the presence of a Salmonella serotype of either one of the groups;
- and
- (b) exposing a further sample of said sample suspected of containing at least one of said Salmonella serotypes to an antibody which specifically binds S. enteritidis but not S. dublin and detecting antibody-antigen specific binding wherein antibody antigen specific binding indicates the presence of S. enteritidis, S. moscow or S. blegdam.
A method of testing a sample for the presence of organisms of the group of Salmonella serotypes expressing SEFA comprising the steps of:
- (a) seeding said sample suspected of containing the organisms into or onto a culture medium supporting the expression of Salmonella enteritidis fimbrial antigen;
- (b) culturing said sample in or on the culture medium;
- and
- (c) exposing a second sample obtained from the culturing step (b) to an antibody which specifically binds to the antigen specifically bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902 or an antibody which specifically binds the epitope bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902 and then detecting antibody-antigen specific binding wherein antibody-antigen specific binding is indicative of the presence of organisms of the group of Salmonella serotypes expressing SEFA.
A method as claimed in 5 wherein culturing in step (b) is carried out at a temperature of at least 22.degree. C. such that SEFA produced uniformly covers the Salmonella cell.
Hybridoma cells deposited at the European Collection of Animal Cell Cultures under accession numbers 90101101 or 90121902.
Monoclonal antibodies produced by the hybridoma cells deposited at the ECACC under accession numbers 90101101 or 90121902.
A test kit for testing a sample for the presence of microorganisms of Salmonella serotypes expressing SEFA comprising:
- (a) hybridoma cells which produce antibodies which specifically bind the antigen specifically bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902 or an antibody which specifically binds the epitope bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902, or
- (b) monoclonal or polyclonal antibodies which specifically bind the antigen specifically bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902 or an antibody which specifically binds the epitope bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902, or
- (c) both (a) and (b).
A test kit for use in a method of testing for the presence of a previous or current infection with Salmonella serotypes expressing SEFA, said test kit comprising SEFA, wherein the SEFA specifically binds an antibody which specifically binds to the antigen specifically bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902 or an antibody which specifically binds the epitope bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902.
An isolated polypeptide comprising SEFA which is specifically bound by an antibody which specifically binds to the antigen specifically bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902 or an antibody which specifically binds the epitope bound by the monoclonal antibody secreted by ECACC 90101101 or ECACC 90121902.
Dependencies are inferred from the claim text.