This test is done to help evaluate a person for hemolytic anemia.
Haptoglobin is a blood protein made by the liver. The haptoglobin levels decrease in hemolytic anemia. Hemolytic anemias include a variety of conditions that result in hemolyzed, or burst, red blood cells.
Decreased values can also indicate a slower type of red cell destruction unrelated to anemia. For example, destruction can be caused by mechanical heart valves or abnormal hemoglobin, such as sickle cell disease or thalassemia.
Haptoglobin is known as an acute phase reactant. Its level increases during acute conditions such as infection, injury, tissue destruction, some cancers, burns, surgery, or trauma. Its purpose is to remove damaged cells and debris and rescue important material such as iron. Haptoglobin levels can be used to monitor the course of these conditions.
Hemoglobin is the protein in the red blood cell that carries oxygen throughout the body. Iron is an essential part of hemoglobin; without iron, hemoglobin can not function. Haptoglobin's main role is to save iron by attaching itself to any hemoglobin released from a red cell.
When red blood cells are destroyed, the hemoglobin is released. Haptoglobin is always present in the blood waiting to bind to released hemoglobin. White blood cells (called macrophages) bring the haptoglobin-hemoglobin complex to the liver, where the haptoglobin and hemoglobin are separated and the iron is recycled.
In hemolytic anemia, so many red cells are destroyed that most of the available haptoglobin is needed to bind the released hemoglobin. The more severe the hemolysis, the less haptoglobin remains in the blood.
Haptoglobin is measured in several different ways. One way is called rate nephelometry. A person's serum is mixed with a substance that will bind to haptoglobin. The amount of bound haptoglobin is measured using a rate nephelometer, which measures the amount of light scattered by the bound haptoglobin. Another way of measuring haptoglobin is to measure it according to how much hemoglobin it can bind.
This test requires 5 mL of blood. The person being tested should avoid taking oral contraceptives or androgens before this test. A healthcare worker ties a tourniquet on the person's upper arm, locates a vein in the inner elbow region, and inserts a needle into that vein. Vacuum action draws the blood through the needle into an attached tube. Collection of the sample takes only a few minutes.
Discomfort or bruising may occur at the puncture site or the person may feel dizzy or faint. Pressure to the puncture site until the bleeding stops reduces bruising. Warm packs to the puncture site relieve discomfort.
Normal results vary based on the laboratory and test method used. Haptoglobin is not present in newborns at birth, but develop adult levels by 6 months.
Decreased haptoglobin levels usually indicates hemolytic anemia. Other causes of red cell destruction also decrease haptoglobin: a blood transfusion reaction; mechanical heart valve; abnormally shaped red cells; or abnormal hemoglobin, such as thalassemia or sickle cell anemia.
Haptoglobin levels are low in liver disease, because the liver can not manufacture normal amounts of haptoglobin. Low levels may also indicate an inherited lack of haptoglobin, a condition found particularly in African Americans.
Haptoglobin increases as a reaction to illness, trauma, or rheumatoid disease. High haptoglobin values should be followed-up with additional tests. Drugs can also effect haptoglobin levels.
Acute phase reactant— A substance in the blood that increases as a response to an acute condition such as infection, injury, tissue destruction, some cancers, burns, surgery, or trauma.
Haptoglobin— A blood protein made by the liver. Its main role is to save iron by attaching itself to any hemoglobin released from a red cell.
Hemoglobin— The protein in the red blood cell that carries oxygen.
Hemolytic anemia— A variety of conditions that result in hemolyzed, or burst, red blood cells.
Normal results vary widely from person to person. Unless the level is very high or very low, haptoglobin levels are most valuable when the results of several tests done on different days are compared.
Örjan Thomas Gunnarson Ouchterlony, a Swedish bacteriologist who was born 1914 in Göteborg (Gothenburg), developed a double immunodiffusion technique in 1948 that, when used in forensics, determines whether a bloodstain is human or animal. This technique is commonly called Ouchterlony double gel diffusion test, which refers to Ouchterlony's critical analysis in 1968 in his Handbook of Immunodiffusion and Immunoelectrophoresis. Another synonym employed is the agar gel immunodiffusion test, AGID.
The binding of an antibody to an antigen is a fundamental reaction of immunology. Antibodies and antigens form complexes that result in the formation of a visible white aggregate, which is called precipitation, making it possible to assay antibody-antigen systems. The antigen in precipitation reactions is soluble and so small that it must combine with many antibodies to form visible clumps. Soluble antigens can be attached to particulate material serving as carriers that can be detected using the more sensitive agglutination technique. Antigen-antibody reactions are widely used in research, laboratory diagnosis of diseases, pregnancy tests, and forensic identification of blood.
The technique involves cutting cylindrical wells into a purified preparation of semi-solidified agar gel in a Petri dish. The wells are filled with antibody or antigen and the dish is allowed to incubate. Homologous antigen and antibody diffuse toward each other from the individual wells to a point in the agar where optimum concentration of each is reached. Subsequently, a precipitin line will form within 18–24 hours somewhere between the two wells. If challenges are mixed together in a single well and allowed to diffuse out into the agar towards the serum test well, multiple precipitin bands are seen routinely. Non-specific reactants diffuse past each other, forming no precipitate. The precipitation reaction is subject to inhibition if either antigen or antibody is present in excess. The qualitative Ouchterlony test can simultaneously monitor multiple antibody-antigen systems and can be used to identify particular antigens.
The Ouchterlony method is wearisome due to the time and interpretative expertise required, and the need for reagent sensitivity and selectivity validation. Today, immunoassay tests are used that rely on immunological principles similar to the Ouchterlony test. Results are accurate, more sensitive, and visible within ten minutes, however, the test apparatus is portable and simple to use, requiring no prior experience to conduct and interpret the results. They can give the crime scene examiner a rapid indication as to whether a sample should be taken for DNA analysis from a bloodstain. Similarly, the laboratory analyst can utilize these tests to confirm whether a bloodstain is of human origin, which may be important where DNA results have failed. If animal blood is suspected, then the Ouchterlony test is utilized.
see also Antibody; Antigen; Blood; Bloodstain evidence; Crime scene investigation; DNA; Homogeneous enzyme immunoassay (EMIT).