An arrangement of enzyme-linked immunosorbent assay to reduce background nonspecific interactions between serum immunoglobulins and B. pseudomallei antigens

Novitskaya I.V., Tereshko D.L., Pechenkina L.A.

Federal Government Health Institution «Volgograd plague Control Research Institute» of the Federal Service for Surveillance in the Sphere of Consumers Rights Protection and Human Welfare, Volgograd, Russia

The aim of the study was to select the conditions for enzyme-linked immunosorbent assay (ELISA) that ensure a reliable decrease in background values of the reaction for detecting serum melioidosis antibodies.

Materials and methods. When performing ELISA, various types of plates, agents blocking the free surfaces of the wells, as well as buffers for diluting samples and antigen-coating were used. The samples for the study were sera of intact and B. pseudomallei-infected laboratory animals, as well as human sera obtained from clinically healthy individuals living in and outside the territory endemic for melioidosis.

Results and discussion. Specific detection of melioidosis antibodies in sera of experimental animals, as well as reduction of background interactions during the study of intact human sera, was achieved by blocking free surfaces of wells with a 5% solution of dry skim milk, as well as using phosphate-buffered saline (pH 7.4) for sensitization of plates and multicomponent Tris-HCl buffer (pH 8.0) containing ethylene glycol, bovine serum albumin, sodium chloride and Tween 20 for sample dilution. The possibility of using the proposed version of ELISA in immunodiagnostics of melioidosis is discussed.