KOOMBS REACTION (R. R. And. Coombs, English immunologist, sort. in 1921; synonym: Koombs's test, antiglobulinovy test) — immunological reaction for identification of monovalent antibodies to auto-and to isoantigens of erythrocytes.
Reaction was developed in 1908 Mr. Moreshi (S. by Moreschi), but was widely used only since 1945 after Koombs showed its role in compatibility test at hemotransfusions, a Rhesus factor conflict, to diagnosis of autoallergichesky and autoimmune states, etc.
To. the river is founded on use in it specially prepared drug — antiglobulinovy serum. In the presence of antiglobulinovy serum the erythrocytes loaded with monovalent antibodies are agglutinated. Erythrocytes, free of on the surface of antibodies, remain not agglutinated.
To. the river is widely applied for: a) establishments of a condition of an isosensitization, i.e. detection of the isoantibodies arising at repeated hemotransfusions (see. Hemotransfusion ) or pregnancies (see. Pregnancy ); b) performance of a compatibility test at hemotransfusions; c) definitions in erythrocytes of a version Rhesus factor (see); d) detection of autoimmune antibodies on erythrocytes of patients of acquired hemolitic anemia (see) and others autoallergichesky diseases (see), and also at some infections proceeding with an allergic component; e) detection of the isoimmune antibodies fixed on erythrocytes of the children suffering hemolitic disease of newborns (see). To. the river is applied also in medicolegal and anthropological researches.
Base material for statement To. rubles are serum or citrate plasma and erythrocytes, the patient. Distinguish two options K. river: indirect and direct. At indirect To. rubles study serum of the patient, in a cut define freely circulating antibodies. At a straight line To. rubles investigate erythrocytes on presence of the antibodies fixed on these uniform elements of blood.
Antiglobulinovy serum for To. rubles receive a lab by immunization. animals (rabbits, goats, sheep, etc.) the globulins emitted from serum of the person with fractionation by ethanol, ammonium sulfate or gel filtering on sephadex. During the receiving antiglobulinovy serum it is necessary to remove the heteroagglutinating antibodies resulting from immunization of animals globulins of the person. Reach it by adsorption of immune serum mix of erythrocytes of people with various blood group [0(1), A (II), V (II), AB (IV)] or cultivation by its isotonic solution of sodium chloride. In the latter case the caption of hetero-agglutinins shall be low (1: 16 — 1: 32), antiglobulinovy serum after cultivation kept good activity in To. the river
Indirect reaction of Koombs
Indirect reaction of Koombs is carried out in two steps. The first phase is completed in the small test tubes of 4 X 0,5 cm in size which are specially intended for this purpose. In each test tube to three drops of serum (integral, 1: 2 etc.), in a cut suspect existence of antibodies, add one drop of a deposit of erythrocytes of the famous antigenic structure. Contents of a test tube are mixed and placed in the thermostat at t ° 37 ° for 1 hour. Then erythrocytes three times wash isotonic solution of sodium chloride. The second stage consists in preparation of 5% of a suspension of the washed erythrocytes and connection on a white (porcelain) plate with a wetted surface of one drop of erythrocytes with one drop of antiglobulinovy serum. The accounting of results is carried out up to 10 min. The exception of false positive results is made by performance of control researches. Use of isotonic solution of sodium chloride instead of antiglobulinovy serum shall not be followed by agglutination of erythrocytes. Performance indirect To. the ruble concerning erythrocytes of the known phenotype of isoantigens allows to establish specificity of antibodies. E.g., a research of serum of the patient with erythrocytes 0(I), CDE, Kk, Fya; 0(1), CDe, Kk, Fua; 0(I), cDe, Kk, Fya; 0(I), cDE, Kk, Fya; 0(I), cde, kk, Fya showed a positive take with a sample of blood in the 1st and 4th cases; other samples of blood (the 2, 3, 5 cases) showed a negative take. Serum contains anti-E-antitela. By means of indirect To. rubles can be revealed monovalent antibodies against antigens: With, D, E, with, e; To, k; Fya, Fyb; Lea, Leb; Jka, Jkb, etc. (see. Blood groups ).
Koombs's straight line reaction
Koombs's Straight line reaction on technology of performance corresponds to the second stage indirect To. river: the studied erythrocytes of the patient which are three times washed by isotonic solution of sodium chloride (5% weigh) connect to antiglobulinovy serum. A straight line To. rubles carry out when there is a basis to believe that erythrocytes of the studied sick in vivo already sensibilizirovana antibodies. A positive straight line To. the river is a diagiostichesky sign at the hemolitic disease of newborns caused by a sensitization of an organism of the woman to antigens of a fruit and penetration of antibodies through a placenta in an organism of the child and also at the acquired hemolitic anemia.
In recent years To. the river is significantly improved. With its help it is possible not only to state existence on erythrocytes of antibodies, but also to establish a class immunoglobulins (see). For this purpose use serum against separate classes of immunoglobulins: IgG, IgM, IgA. Isoimmune antibodies against a Rhesus factor, Kell-antigenov, Daffi-antigenov and other antigens, and also thermal autoimmune antibodies, as a rule, belong to IgG. Cold-reactive autoimmune antibodies, and also isoimmune antibodies against Le and some other antigens, as a rule, belong to IgM. Only seldom found autoimmune antibodies have the nature of IgA (see. Autoantibodies ) .
Bibliography: Dygin V. P. Autoimmune diseases in clinic of internal diseases, L., 1970, bibliogr.; The cashier-with to and y I. A. and Alekseev G. A. Clinical hematology, M., 1970; P. N Jambs. From antigens and from an antibody of the person also pathologies, M., 1974, bibliogr are normal.; In about i v i n P. e. a. Les anemies hemolytiques, p. 93, P., 1971, bibliogr.; Clinical aspects of immunology, ed. by P. G. H. Geli and. lake, Oxford, 1975; Coombs R. R. A., M o u of a n t A. E. a. Race R. R. In-vivo isosensitisation of red cells in babies with haemolytic disease, Lancet, v. 1, p. 264, 1946.
E. A. Zotikov.