Four EIA (enzyme immunoassay) test systems for the detection of salmonella O-antigens in various biological tissues were studied. To find the optimum test system two types of affinity purified antibodies were used to coat the microtitre plates: (i) monospecific antibodies isolated on immunoadsorbent bearing synthetic O-antigen factor 4 (O:4; salmonella serogroup B) as ligand, (ii) antibodies specific for lipopolysaccharide (LPS) B isolated from the IgG fraction of hyperimmune sera. The use of affinity purified antibodies led to an increase in the sensitivity of 'sandwich' EIA by an order of magnitude and to improved specificity. Competitive EIA was ten to twenty times less sensitive. It is demonstrated for the first time that salmonella O-antigens can be detected in the sera of animals within a day after challenge. For patients with salmonellosis, O-antigen could be detected after the fifth day of illness, but in the urine and faeces only (not in the blood serum), in 30%-70% of cases. This substantially improves the identification of salmonellosis from among other enteric infections. In competitive EIA, inhibition of standard antibodies by the sera under study was caused not by the presence of O-antigen but by antibodies homologous to the coating antigen. This resulted in "blocking' of the latter and led to false-positive results. The results obtained enable the optimum EIA technique to be selected to improve the serological diagnosis of salmonellosis with both synthetic salmonella O-antigens and LPS.
Система генетического контроля реакции врожденного иммунитета на гриппозную инфекцию и функции генов позволяет вести разработку системного лечения гриппа с ориентацией на фенотипические проявления мутаций с учетом наследственной предрасположенности индивида к тяжелому течению заболевания и/или развитию осложнений.
Aim. To determine the potentialities of use of affinity interaction of immobilized biologically active
substances (bacterial cells or their fragments, toxins, antigens of various chemical nature, immunoglobulins, enzymes, gangliosides, etc.) for medical practice.
Materials and methods. Emulsion polymerization of acrylamide monomers in the gaseous nitrogen
current was used as a basic method for preparation of solid-phase magnetic immunosorbents (MIC). A
procedure for preparation of siliceous MIC was also applied. The prepared MICs were used a solid
phase in enzyme immunoassay and immunofluorescence assay and the recorded data were compared
with those of stidied conventionally used in practical medicine.
Results. The use of MIC made it possible to detect pathogens of particularly dangerous infections in
large volumes of the samples contaminated with another microflora. With the proposed MIC, one can
stand a good chance of surveying large contingents of the population, of obtaining the quantitative results in shorter periods to establish a diagnosis. With this, the sensitivity and specificity of immunoassays substantially increase. Whether MIC may be used as selective hemosorbents to remove specific antibodies from the blood of patients with rheumatic diseases for therapeutic purposes was studied.
Conclusion. The findings are indicative of wide potentialities of use of affinity interaction of biologically active substances immobilized on inert carriers with the inserted magnetic material in the laboratory
diagnosis of diseases of both infectious and autoimmune nature, which may be widely used in the inand outpatient settings.
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