250-0020s Simulated ABO & Rh blood typing

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                                                                                    Simulated ABO & Rh
                                                                                       Blood Typing
                                                                                        Lab Activity
                                                                                      Student Study Guide

                                                                                               BACKGROUND

                                                                     Around 1900, Karl Landsteiner discovered that there are at least four
                                                                     different kinds of human blood, determined by the presence or ab-
                                                                     sence of specific agglutinogens (antigens) on the surface of red blood
                                                                     cells (erythrocytes). These antigens have been designated as A and B.
                                                                     Antibodies against antigens A or B begin to build up in the blood
                                                                     plasma shortly after birth, the levels peak at about eight to ten years
                                                                     of age, and the antibodies remain, in declining amounts, throughout
    Agglutinogens (Antigens):                                        the rest of a person's of life. The stimulus for antibody production is
    Agglutinogens are substances                                     not clear; however, it has been proposed that antibody production is
    found on the surface of                                          initiated by minute amounts of A and B antigens that may enter the
    erythrocytes.                                                    body through food, bacteria, or other means. Humans normally pro-
                                                                     duce antibodies against those antigens that are not on their erythro-
                                                                     cytes: A person with A antigens has anti-B antibodies; a person with
    Agglutinins (Antibodies):                                        B antigens has anti-A antibodies; a person with neither A nor B anti-
    Agglutinins are antibodies                                       gens has both anti-A and anti-B antibodies; and a person with both A
    found in plasma.                                                 and B antigens has neither anti-A nor anti-B antibodies (Figure 1).
                                                                     Blood type is based on the antigens, not the antibodies, a person pos-
                                                                     sesses.

                                                                     The four blood groups are types A, B, AB, and O. Blood type O, char-
                                                                     acterized by the absence of A and B agglutinogens, is the most com-
                                                                     mon in the United States and is found in 45% of the population. Type
                                                                     A is next in frequency, and is found in 39% of the population. The fre-
                                                                     quencies at which types B and AB occur are 12% and 4% respectively.


                                                                                  Figure 1
                                       Antigens on                              Antibodies
                       Blood                                                                          Can Give      Can Receive
                                       Erythrocytes                             in Plasma
                       Type                                                                           Blood To      Blood From
                                     (Agglutinogens)                           (Agglutinins)
                           A                       A                              Anti-B                A, AB             O, A

                           B                       B                              Anti-A                B, AB             O, B

                          AB                 A and B                     Neither Anti-A nor Anti-B       AB          O, A, B, AB

                           O          Neither A nor B                      Both Anti-A and Anti-B    O, A, B, AB           O



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                                                                                             ABO System
                                                                                        Process of Agglutination
                                                                     There is a simple test performed with antisera containing high levels
                                                                     of anti-A and anti-B agglutinins to determine blood type. Several
                                                                     drops of each kind of antiserum are added to separate samples of
                                                                     blood. If agglutination (clumping) occurs only in the suspension to
                                                                     which the anti-A serum was added, the blood type is A. If agglutina-
                                                                     tion occurs only in the anti-B mixture, the blood type is B. Agglutina-
                                                                     tion in both samples indicates that the blood type is AB. The absence
                                                                     of agglutination in any sample indicates that the blood type is O
           DID YOU KNOW?                                             (Figure 2).
      The average life span of a red
      blood cell is about 120 days.
                                                                                                      Figure 2
                                                                                 Agglutination Reaction of ABO Blood-Typing Sera
                                                                                                Reaction
                                                                                                                                  Blood Type
                                                                                Anti-A Serum           Anti-B Serum
                                                                                Agglutination         No Agglutination                  A
                                                                               No Agglutination        Agglutination                     B
                                                                                Agglutination          Agglutination                    AB
                                                                               No Agglutination       No Agglutination                  O


                                                                                     Importance of Blood Typing
                                                                     As noted in the table above, people can receive transfusions of only
                                                                     certain blood types, depending on the type of blood they have. If in-
                                                                     compatible blood types are mixed, erythrocyte destruction, agglutina-
                                                                     tion and other problems can occur. For instance, if a person with type
                                                                     B blood is transfused with blood type A, the recipient's anti-A anti-
                                                                     bodies will attack the incoming type A erythrocytes. The type A
                                                                     erythrocytes will be agglutinated, and hemoglobin will be released
                                                                     into the plasma. In addition, incoming anti-B antibodies of the type A
                                                                     blood may also attack the type B erythrocytes of the recipient, with
           DID YOU KNOW?                                             similar results. This problem may not be serious, unless a large
      Donor blood contains only                                      amount of blood is transfused.
      packed red blood cells. There
      is no plasma in donor blood,                                   The ABO blood groups and other inherited antigen characteristics of
      thus there are no antibodies                                   red blood cells are often used in medico-legal situations involving
      present.                                                       identification of disputed paternity. A comparison of the blood
                                                                     groups of mother, child, and alleged father may exclude the man as a
                                                                     possible parent. Blood typing cannot prove that an individual is the
                                                                     father of a child; it merely indicates whether or not he possibly could
                                                                     be. For example, a child with a blood type of AB, whose mother is
                                                                     type A, could not have a man whose blood type is O as a father.




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Copymaster. Permission granted to make unlimited copies for use in any one
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                                                                                        The Genetics of Blood Types
                                                                     The human blood types (A, B, AB, and O) are inherited by multiple
                                                                     alleles, which occurs when three or more genes occupy a single locus
                                                                     on a chromosome. Gene IA codes for the synthesis of antigen
                                                                     (agglutinogen) A, gene IB codes for the production of antigen B on the
                                                                     red blood cells, and gene i does not produce any antigens. The pheno-
                                                                     types listed in the table below are produced by the combinations of
                                                                     the three different alleles: IA, IB, and i. When genes IB and IA are pre-
                                                                     sent in an individual, both are fully expressed. Both IA and IB are
          DID YOU KNOW?                                              dominant over i so the genotype of an individual with blood type O
     Camels and their relatives are                                  must be ii (Figure 3).
     the only mammals having
     oval red blood cells.                                                                               Figure 3
                                                                                  Phenotype                  Possible Genotypes
                                                                                                                       IAIA
                                                                                        A
                                                                                                                       IAi
                                                                                                                       IBIB
                                                                                         B
                                                                                                                       IBi
                                                                                       AB                              IAIB
                                                                                        O                              ii

                                                                               Use IA for antigen A, IB for antigen B, and i for no antigens present.
                                                                               Genes IA and IB are dominant over i.
                                                                               AB blood type results when both genes IA and IB are present.


                                                                                                         Rh System
                                                                     In the period between 1900 and 1940, a great deal of research was
                                                                     done to discover the presence of other antigens in human red blood
                                                                     cells. In 1940, Landsteiner and Wiener reported that rabbit sera con-
                                                                     taining antibodies for the red blood cells of the Rhesus monkey
                                                                     would agglutinate the red blood cells of 5% of Caucasians. These anti-
                                                                     gens, six in all, were designated as the Rh (Rhesus) factor, and they
                                                                     were given the letters C, c, D, d, E, and e by Fischer and Race. Of
                                                                     these six antigens, the D factor is found in 85% of Caucasians, 94% of
           DID YOU KNOW?                                             African Americans, and 99% of Asians. An individual who possesses
     Rh is so named because the                                      these antigens is designated Rh+; an individual who lacks them is
     initial study was done with                                     designated Rh-.
     Rhesus monkeys.
                                                                     The genetics of the Rh blood group system is complicated by the fact
                                                                     that more than one antigen can be identified by the presence of a
                                                                     given Rh gene. Initially, the Rh phenotype was thought to be deter-
                                                                     mined by a single pair of alleles. However, there are at least eight al-
                                                                     leles for the Rh factor. To simplify matters, consider one allele: Rh+ is
                                                                     dominant over Rh-; therefore, a person with an Rh+/Rh- or Rh+/Rh+
                                                                     genotype has Rh+ blood.



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                                                                     The anti-Rh antibodies of the system are not normally present in the
                                                                     plasma, but anti-Rh antibodies can be produced upon exposure and
                                                                     sensitization to Rh antigens. Sensitization can occur when Rh+ blood
                                                                     is transfused into an Rh- recipient, or when an Rh- mother carries a
                                                                     fetus who is Rh+. In the latter case, some of the fetal Rh antigens may
                                                                     enter the mother's circulation and sensitize her so that she begins to
                                                                     produce anti-Rh antibodies against the fetal antigens. In most cases,
                                                                     sensitization to the Rh antigens takes place toward the end of preg-
                                                                     nancy, but because it takes some time to build up the anti-Rh antibod-
                                                                     ies, the first Rh+ child carried by a previously unsensitized mother is
                                                                     usually unaffected. However, if an Rh- mother, or a mother previ-
                                                                     ously sensitized by a blood transfusion or a previous Rh+ pregnancy,
                                                                     carries an Rh+ fetus, maternal anti-Rh antibodies may enter the fetus'
                                                                     circulation, causing the agglutination and hemolysis of fetal erythro-
                                                                     cytes and resulting in a condition known as erythroblastosis fetalis
                                                                     (hemolytic disease of the newborn). To treat an infant in a severe
                                                                     case, the infant's Rh+ blood is removed and replaced with Rh- blood
                                                                     from an unsensitized donor to reduce the level of anti-Rh antibodies.

                                                                                          Blood Components
                                                                     The formed elements in blood include erythrocytes, or red blood cells
                                                                     (RBCs); various types of leukocytes, or white blood cells (WBCs); and
                                                                     platelets.

          DID YOU KNOW?                                              Erythrocytes are circular, biconcave disks of 5 to 8 micrometers. Their
     Leukocytes are capable of                                       chief function is to transport oxygen (O2) and carbon dioxide (CO2).
     amoeboid movement and are                                       The transport of O2 and CO2 depends largely on the hemoglobin pre-
     sometimes called amoebo-                                        sent in the erythrocytes. The biconcave shape is also related to the
     cytes.                                                          erythrocytes function of transporting gases, in that it provides an in-
                                                                     creased surface area through which gases can diffuse.

                                                                     The number of circulating RBCs is closely related to the blood's oxy-
                                                                     gen-carrying capacity. Any changes in the RBC count may be signifi-
                                                                     cant. RBC counts are routinely made to diagnose and evaluate the
                                                                     course of various diseases.

                                                                     Leukocytes range in size from approximately 9 to 25 micrometers and
                                                                     function primarily to control various disease conditions. Leukocytes
                                                                     can move against the current of the bloodstream through amoeboid
                                                                     movement, and pass through the blood vessel walls to enter the tis-
                                                                     sues. The total WBC count normally varies from 5,000 to 10,000/mm3.
                                                                     Certain infectious diseases are accompanied by an increase in WBCs.
                                                                     If the number exceeds 10,000/mm3, the person has an acute infection.
                                                                     If it drops below 5,000/mm3, the person may have a condition such
                                                                     as measles or chicken pox. The percentage of the different types of
                                                                     leukocytes present in the blood may also change in particular dis-
                                                                     eases, this number is important for diagnostic purposes and is called
                                                                     a differential count.




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Copymaster. Permission granted to make unlimited copies for use in any one
school building. For educational use only. Not for commercial use or resale.
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                                                                                                    OBJECTIVES

                                                                              Define agglutinogen and agglutinin
                                                                              Perform an actual blood typing procedure
                                                                              Observe the antigen/antibody reaction in simulated blood
                                                                              Determine the ABO and Rh blood type of four unknown
                                                                               samples
                                                                              Prepare a wet mount of simulated blood
                                                                              Estimate the number of erythrocytes and leukocytes in
                                                                               normal blood
                                                                              Understand requirements for blood transfusions

                                                                                                    MATERIALS

                                                                     MATERIALS NEEDED PER GROUP

                                                                     4         Blood typing slides
                                                                     12        Toothpicks
                                                                     1         Microscope slide
           DID YOU KNOW?                                             1         Coverslip
    Prior to the mid-1500s, no one                                             Compound microscope (400X magnification)
    had any conception of blood                                                Marker
    circulating through the organs
    in the body.                                                     SHARED MATERIALS

                                                                     4         Unknown blood samples:
                                                                                  Mr. Smith
                                                                                  Mr. Jones
                                                                                  Mr. Green
                                                                                  Ms. Brown
                                                                               Simulated Anti-A Serum
                                                                               Simulated Anti-B Serum
                                                                               Simulated Anti-Rh Serum

                                                                                                       PROCEDURE

                                                                                        Although WARD'S Simulated Blood is completely safe, non-
                                                                                        biological, and non-toxic, you should wear the proper personal
                                                                                        protective equipment to mimic the experience of an actual he-
                                                                                        matology laboratory.

                                                                     PART A: ABO and Rh BLOOD TYPING

                                                                     1.        Label each blood typing slide:
                                                                                  Slide #1: Mr. Smith
                                                                                  Slide #2: Mr. Jones
                                                                                  Slide #3: Mr. Green
                                                                                  Slide #4: Ms. Brown

                                                                                                             2002 WARD'S Natural Science Establishment, Inc.
Copymaster. Permission granted to make unlimited copies for use in any one
school building. For educational use only. Not for commercial use or resale.
                                                                                            5                            All Rights Reserved
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