General characteristic of Rh-Hr system.

The Agglyutinogen system a rhesus (Rh-Hr) after antigens of ABO has the greatest value for clinical practice. In transfusiology 6 antigens have the greatest clinical value.

For their designation use two nomenclatures. According to the nomenclature of Wienera offered in 1942 antigens a rhesus are designated by symbols: Rh0, Rh', Rh'', Rh1w, Hr', Hr''.

Other nomenclature offered in 1944 to Fischer and R. Race uses alphabetic references: D, C, E, Cw, c, e. Antigens a rhesus, as well as other group signs of a blood of the person, are inherited from parents and during life don't change. Antigens a rhesus are in a membrane of erythrocytes. Antigens of D,C, E are inherited on dominant type, and antigens of c and e – on recessive.

In this regard it is possible to distinguish two blood types from recipients: Rh-positive (Rh +) to which the blood of all persons having an antigen of D in erythrocytes (simply concerns: DCE, DSE, DSE, Dce), and Rh-negative (Rh-) to which the blood of all persons which don't have an antigen of D belongs (simply dCE, dCe, dcE, dce).

Definition of a blood type by means of tsoliklon

Monoclonal anti-A and anti-B antibodies are produced by two various mouse hybridomas and belong to class M immunoglobulins.

Tsoliklon anti-AB represents an admixture monoclonal anti-A and anti-B antibodies.

Monoclonal antibodies give faster and more expressed agglutination test with agglutinogens A or B, and the result of their interaction can be considered in 3 minutes.

Definition of a blood type is made by method of a direct hemagglutination on the plane: on a plate or on the tablet indoors with good lighting at a temperature of +15-+25 degrees Celsius.

A. On the tablet or on a plate individual pipettes apply tsoliklona anti-A and anti-B on one big drop (0,1 ml) under the corresponding inscriptions.

B. Near drops of antibodies put on one small drop of the studied blood (0,02-0,03 ml) and admix a blood with reagent.

C. The plate or the tablet need to be shaken slightly within three minutes. Agglutination of erythrocytes from tsoliklona usually comes in the first 3-6 seconds, but it is necessary to conduct observation three minutes in view of later emergence of agglutination with the erythrocytes containing weak kinds of antigens A.

G. The result of reaction can be positive or negative. The positive take is expressed in agglutination of erythrocytes. At negative reaction the drop remains evenly painted in red color, agglyutination it aren't found.

Definition of a blood type in the two-dimensional way

A. Definition of a blood type consists in the two-dimensional way in simultaneous definition of group agglutinogens in erythrocytes of an blood on tsoliklona and group agglutinins in Serum of the studied blood by means of standard erythrocytes.

B. For definition of a blood type in the two-dimensional way except monoclonal antibodies anti-A, anti-B and anti-AB use standard erythrocytes of O(I), A (II) and B (III) group.

B. The blood for mixing is taken from a vein or a place of a nyxis of a finger in a dry pure test tube. The blood is centrifuged or left alone for 20-30 minutes for unit of Serum.

G. Definition is made on a white plate on which top part apply designations from left to right: anti-A, Anti-B. At the upper edge inscribe a surname and initials of the person at whom define a blood type.

D. Under the corresponding designations of blood types apply on a plate on one big drop (0,1 ml) of standard monoclonal antibodies.

E. Under designations of O(I), A (II) and B (III) apply on the right part of a plate on one small (0,01 ml) to a drop of standard erythrocytes in the following order from left to right: O (I), A(II) and B (III).

Zh. From the test tube containing the patient's blood, a pipette extract Serum and drop it on one big (0,1 ml) a drop on the prepared standard erythrocytes. After that the same pipette gather erythrocytes of an testing blood since a bottom of a test tube and put them on small (0,01 ml) to a drop near each drop of the prepared monoclonal antibodies.

H. In all drops of an antibody and Serum carefully mix with erythrocytes, using glass rods, the plate is shaken, then for 1-2 minutes left alone and again periodically shaken. Observation over the course of reaction is made by not less than five minutes.

I. In process of offensive of agglutination with standard erythrocytes, but not earlier than in 3 minutes, in those drops, in which it I came, add on one drop (0,05 ml) of isotonic NaCl solution and continue observation when rocking a plate before the efflux of five minutes.

The accounting of reaction is made by comparison of the results received by means of standard Serums and erythrocytes.

Results of the reactions received by means of monoclonal antibodies and standard erythrocytes have to coincide, i.e. point to the maintenance of the agglutinogens and agglutinins corresponding to the same blood type. These results can be expressed in four various combinations.

A. The studied blood didn't give agglutination with monoclonal antibodies that points to absence in it group agglutinogens and belonging to O(I) group. Thus Serum of the studied blood (the lower series) gives negative reaction with standard erythrocytes of O(I) group and positive – with erythrocytes of A(II) and B groups (III). It indicates existence in an blood of agglutinins α and b, i.e. confirms its belonging to O(I) group.

B. By means of monoclonal antibodies anti-A in the studied blood existence of an agglutinogen is established A. Thus Serum of the studied blood doesn't give agglutination with standard erythrocytes of O(I) and A groups (II), i.e. it doesn't contain agglutinin a, but agglutinates erythrocytes of B(III) group, i.e. in its structure there is an agglutinin b that points to belonging of an testing blood to group A (II) b.

B. By means of monoclonal antibodies anti-B in the studied blood existence of an agglutinogen of B is defined. Thus Serum of the studied blood gives negative reaction with standard erythrocytes of O(I) and B groups (III) that confirms lack of agglutinin b, and positive with erythrocytes of A(II) group that speaks about availability of agglutinin an and confirms belonging of an blood to group B (III) α.

G. By means of monoclonal antibodies anti-A and anti-B in the studied blood existence of agglutinogens A and B. In this case as control conduct research with tsoliklony anti-AB. If to it also there was an agglutination, carry out assay with a normal saline solution of chloride sodium lack of agglutination with which confirms specificity of reaction. Thus Serum of the studied blood gives negative reaction with standard erythrocytes of all three groups that specifies on lack of agglutinins in the studied blood, i.e. confirms belonging of an blood to AB (IV) group of the lake.