COMPARATIVE ANALYSIS OF EFFECTIVENESS OF SOLID-PHASE METHODS OF IMMUNE DETECTION OF BOTULINIC TOXIN IN BLOOD SERA OF A PATIENT WITH BOTULISM DIAGNOSIS

Abstract

Aim. Comparison of effectiveness of solid phase methods of immune detection of botulinic toxin in blood sera of a patient with botulism diagnosis: dot-immune assay using specific anti-botulinic antibodies (AT) labeled with nanoparticles of colloid silver, phosphorescent analysis (PHOSPHAN) using streptavidin label with platinum coproporphyrin (PtCP) and polystyrene nanoparticles, containing chelate complex of europium ions with naphthoyl trifluoroacetone (NA-Eu). Materials and methods. Silver nanoparticle labeled IgG isolated from a commercial diagnostic polyvalent sera against type А, В, С, E, F botulotoxins manufactured by SPA Allergen (Stavropol) with 5000 - 10000 IU activity and biotin conjugated commercial monoclonal antibodies against botulotoxin A, polyclonal mono-specific AB against botulotixin В and E and polyvalent immunoglobulin against botulotoxin А, В, С, E, F. Detection ofbotulotoxin in clinical material was carried out in dot-immunoassay on nitrocellulose membrane by PHOSPHAN method in an experimental test system using 2 detector systems based on streptavidin: PtCP and NA-Eu. Results. Botulotoxin was detected in blood sera of the botulism patient using both of the developed immune detection methods. PHOSPHAN method allowed to identify serotype В botulotoxin, that corresponded with the results obtained in botulotoxin biological neutralization reaction. Sensitivity of PHOSPHAN with NA-Eu luminescent nanoparticle based detection system was higher than with PtCP label. Conclusion. The developed methods (PHOSPHAN and dot-immunoassay) differ by high specificity and sensitivity and may be recommended for express detection of botulinic toxin in clinical material.