[The application of monoclonal peroxidase conjugates to identify comma bacillus of serum groups 01 and 0139 in the reaction of dot-immune analysis].

The source of monoclonal antibodies was chosen the cultural fluid of hybridoma-producers deposited in the specialized collection of cell cultures of vertebrates (St. Petersburg) with numbers RKKK(P) 386D and RKKK(P) 674D. The specific immunoglobulin (Ig) from cultural fluid was concentrated by precipitation with saturated solution of ammonium sulfate. The scheme of obtaining monoclonal antibodies included activation of peroxidase, conjugation of activated peroxidase with Ig, removal of unbounded proteins, storage and control. The preservation of activity of conjugates was supported with BSA (10%) or glycerin (50%). The last on is preferable to be applied for this purpose. The test of monoclonal antibody-01 and monoclonal antibody-0139 of peroxidase conjugates with kit of strains of comma bacillus 01 and 0139 demonstrated their strict specificity because they interacted only with corresponding serum groups under absence of crossed reactions with representatives of geterologic microorganisms. The direct dot-immune analysis is carried out during 1.5 hour and its sensitivity is within the limits 105-106. The application of diagnostic monoclonal peroxidase conjugates 01, 0139 in laboratory practice can promote the increase of specificity of serologic analysis of cholera and saving time-frame of its application.

[The optimization of conditions of diagnostic fluorescent monoclonal immunoglobulins production to identify comma bacillus of serogroups O1 and O139].

The article considers, the issue of producing the species-specific fluorescent monoclonal immunoglobulins to detect comma bacillus of serogrups O1 and O139 in the reaction of direct immunfluorescence. It is established that not only ascitic but culture fluid too can be the source of monoclonal antibodies for producing fluorescent conjugates. The optimal conditions are selected to produce the fluorescent monoclonal immunoglobulins-monoclonal antibodies. The corresponding producing techology can be reproduced at any time in view of availability of hybrid-producers of monoclonal antibodies O1 and monoclonal antibodies O139 in the institute cryodepositoty. The results of testing the fluorescent preparations on homologous and heterologous strains demonstrated their strict specificiy and high sensibility regarding comma bacillus of serogroups O1 and O139. The new preparations favor significant increase of effectiveness of diagnostics of V. cholerae O1 and O139.

[Identification of Vibrio cholerae serogroups O1 and O139 by monoclonal antibodies in the slide agglutination test].

The agglutinating properties of MCA-O1 of the IgG class and MCA-O139 of the IgM class towards epitopes of O-antigen of Vibrio cholerae O1 and accordingly Vibrio cholerae O139 were studied. The ascitic and cultural fluids by hybridomas F8G12 and D11 deposited in the specialized Collection of Cell Cultures of Vertebrates (Saint Petersburg) under RKKK (II) 386 D and RKKK (II) 674 D were the sources of monoclonal immunoglobulins. The advantage of diagnostic monoclonal immunoglobulins is that they are distinguished for strict specificity and their use in practical health care contributes to the higher specificity of a laboratory test for cholera and to its shorter performance.