METHOD OF DETECTING AN IMMUNOLOGICAL REACTION

摘要

Both blood group and non-blood group immunological reactions are the subject of this invention. A method of forward grouping is presented wherein known antibodies are attached directly to a solid surface and the red blood cells presented for immunological reaction are activated by treatment with a proteolytic enzyme. A reverse blood grouping procedure according to the invention utilizes synthetic or purified antigens which may also be attached directly to a solid surface. The surface is then contacted with an unknown blood component to permit antibodies to undergo immunological reaction with the previously attached antigens. A solution of red blood cells is used as the indicator mechanism and is activated before usage by treatment with a proteolytic enzyme. The method of the invention is applicable to non-blood group immunological reactions wherein purified antigens or antibodies or synthetic substances are attached directly to a solid surface. The surface is then contacted with an unknown fluid that may contain antigens or antibodies specific to those previously attached. A solution of red blood cells including the known antigen or antibody attached to the solid surface is then brought into contact so that the latter will undergo an immune reaction and indicate the presence of specific antigens or antibodies in the unknown fluid. The invention encompasses a method of performing a major crossmatch or compatibility test utilizing anti-human immunoglobulin that is attached directly or indirectly to a solid surface which is then contacted by the two blood components which have been previously incubated to permit an antigen-antibody interaction of red blood cells and antibodies. The antibody sensitized cells will immunologically adhere to the solid phase. In an alternative major crossmatch procedure, a binder is used to attach red blood cell membranes from a blood donor and the serum from a recipient is allowed to undergo an immune reaction with these membranes on the solid surface. They are then contacted with red blood cells or synthetic particles sensitized with anti-human immunoglobulin. Antibody screening and antibody identification are carried out by attaching known antigen carrying cells to a solid surface. Optionally, these membranes can be preserved with a dilute glutaraldehyde solution. The solid surface is then contacted with the unknown solution which will undergo an immune reaction to the extent antibodies specific to the previously adhered antigens are present. Red blood cells or synthetic particles coated with anti-human immunoglobulins are used as the indicator mechanism. These cells or particles will undergo an immune reaction to the extent antibodies in the unknown blood component have immunologically reacted with the antigens on the solid surface. A direct anti-globulin test may be performed by attaching anti-human immunoglobulin to the solid surface and then subjecting the solid surface to the unknown blood component which may contain immunoglobulin coated red blood cells.