Hemolyzed Information and Courses from MediaLab, Inc.

Band Appearance Possible Cause Short Migration Patterns Contaminated or Aged Buffer Diffuse Bands Markedly Wet Gels Poor Sample Application Streaks Perpendicular to Bands Tearing/Poking Gel in Sample Application Weak Bands Not Enough Sample Not Enough Stain Other Causes of Irregular and Distorted Bands Hemolyzed Sample Bent or Dirty Applicator Air Bubbles in Sample Application Too Much Sample Wick Flow Too Much Heat or Drying

Specimen rejection criteria established by your laboratory should be followed at all times, as improperly collected or processed coagulation specimens could adversely affect patient results. Generally speaking, hemolyzed specimens should not be used in coagulation testing because ADP liberated from lysed red blood cells can interfere with a number of coagulation tests, especially those involving platelet assessment. Grossly lipemic specimens may cause erroneous results or a clot may not be detected if a photo-optical coagulation system is used. An alternative method that is not affected by lipemia, such as an electromechanical method, may be required One way to avoid a grossly lipemic specimen is to ask the patient to fast prior to specimen collection.

Serum iron (SI) is a measure of circulating iron bound to transferrin and is reflective of total body iron. SI is elevated in hereditary hemochromatosis (HH) and acute hepatitis. SI is decreased in iron deficiency anemia and chronic inflammation. SI concentrations exhibit diurnal variation, with the lowest values occurring around midnight. In addition, specimens collected from the same individual at the same time of the day may exhibit day to day variations as high as 40%. SI determinations are also affected by diet, menstrual cycle, pregnancy, ingestion of iron supplements, and oral contraceptive use. SI levels alone are considered insensitive indicators of HH. SI is typically measured on automated analyzers using spectrophotometric methods. Iron in the sample is released from transferrin with an acid reagent, reduced to the ferrous state, and reacted with a chromogen such as bathophenanthroline or ferrozine. The intensity of the color change is proportional to the iron concentration. Interference can arise from the use of a hemolyzed sample and contamination of reagents and water with iron. A typical reference interval for SI is 60 - 150 micrograms/dL. SI is usually ordered along with its companion test, the total iron binding capacity (TIBC), or with transferrin (Tf).(2)

Antibodies of the ABO system cause agglutination of saline-suspended red cells at 4°C to 20°C. Heating to 37° weakens the reaction. “Naturally” occurring ABO antibodies may not be strong enough to agglutinate cells without centrifugation. Thus, testing serum for the presence of anti-A or anti-B has classically been performed using the tube system in which serum and cells added to a test tube are centrifuged and then evaluated for agglutination. A slide test has also been performed for forward reactions. Although tube tests are still in wide use, newer systems utilizing other technology such as gel agglutination are becoming more prevalent. The image on this page illustrates agglutination reactions observed with the tube system, from 4+ in the topmost image, to 0 in the lowest image. ABO reactions should be strong. Weak or missing reactions occur, but must be "resolved" before blood products can be released.4+ agglutination: Red blood cell button is a solid agglutinate; clear background.3+ agglutination: Red blood cell button breaks into several large agglutinates; clear background.2+ agglutination: Red blood cell button breaks into many medium-sized agglutinates; clear background; no free red blood cells.1+ agglutination: Red blood cell button breaks into many small clumps barely visible macroscopically; background is turbid; many free red blood cells.Negative: No agglutinated red blood cells present; red cells are observed flowing off the red blood cell button during the process of grading.Other reaction which may occur are the mixed-field reaction, in which mixtures of agglutinated and unagglutinated red blood are present; and hemolysis, in which red cells are hemolyzed by the antibody. Both of these patterns are considered positive reactions.

Medical errors also occur in the analytic processes and systems of patient care. Analytic errors begin with problems in the transportation of medical samples for testing. These occur between the patient's location and the testing facility. They happen during the time between specimen collection and arrival in the testing facility. The possibility for analytic medical error continues through the analytic processes and procedures of medical testing. Analytic medical error also includes systems, processes, and procedures involved in the transmission and reporting of test results. These medical errors occur during the time the laboratory is directly involved in receiving, analyzing, and reporting test samples. Examples: Wrong transport storage or temperature Delay in transport Sample mixup during transport Acceptance of unacceptable samples that are insufficient, hemolyzed, or clotted Centrifugation, mixing, and other test sample preparation errors Wrong test procedures Test control errors Sample mixup during testing Outdated reagents Wrong reagents Test result mixup Transcription errors Data reporting process errors Result report delays

Hemolysis means the breakup of fragile red blood cells within the specimen, and the release of their hemoglobin (the red oxygen carrying substance present within the red cells), and other substances, into the plasma.A hemolyzed specimen is one which has undergone hemolysis. A hemolyzed specimen can be recognized after it is centrifuged by the red color of the plasma.

Pre-analytical errors are errors that occur prior to the testing process. Hemolyzed specimens, clotted specimens, incorrect tube type, and inadequate tube fill can all produce pre-analytical errors. Fortunately, many of these errors can be detected by the laboratory analyst so that corrections can be made before testing begins or before resulting and reporting the test. Unfortunately, the correction that needs to be made usually involves redrawing the patient. The table on the following page lists several preanalytical errors that can occur during the phlebotomy procedure.

Red cell casts appear as clear cylinders containing red blood cells and may have an orange red tinge. Their presence indicates bleeding into the nephron. Red cells within the cast are rapidly hemolyzed and the cast becomes a hemoglobin cast, having an orange color and a homogeneous ground-glass texture. In order for a cast to be considered an RBC cast, the outline of the red cells must be clearly visible in at least one area of the cast.