STUDY OF ANTIBODIES TO PATHOGENIC FACTORS IN STAPHYLOCOCCAL IMMUNOGLOBULIN

Abstract

            F(ab’) fragments of antibacterial, as well as alpha-toxin, PV-leukocidin and hyaluronidase antibodies in polyclonal staphylococcal immunoglobulin have been studied, the specific activity of which was investigated by enzyme-linked immunoenzyme assay (ELISA) and passive (indirect) hemagglutination reaction using appropriate test systems. F(ab’) fragments were obtained using a new technology using immobilized pepsin. To obtain the F(ab’)2 fragment we used a special kit produced by Thermo Scientific (USA) (PierceTM Fab Preparation Kit), which contains all the components necessary for obtaining the F(ab’)2 fragment of the antibody and is a fast and effective means of obtaining and purifying the said product. The titer of antibodies to α-toxin was determined in the Lh (limes-haemolisis) reaction, which was 150 IU/ml; the range of antibacterial antibody titers in immune serum and immunoglobulin in the passive hemagglutination test was -1:3200 - 1:12800; the titer of antileukocidin antibodies was -1:640 - 1:1280, and to hyaluronidase - 1:320 - 1:640. These indicators in normal serum ranged within 1:10-1:40. Antibodies in antistaphylococcal immunoglobulin solution were determined by the immunoenzyme method, the results of which were registered on an immunoenzyme reader (Sunostik SPR-960). The average data of normal (control) sera for staphylococcal toxin was 0.081; while the positivity index of immune serum and immunoglobulin for the said toxin was 10.08. The positivity index for antibacterial antibodies was 9.2517, the positivity index for PV-leukocidin was 4.3968, and for hyaluronidase - 0.9214.