Intended Use
For the determination of total complement activity (CH50) in human serum. This assay is used to determine the functional integrity of the entire classic complement pathway.
For In Vitro Diagnostic Use.
Summary
The complement system consists of a group of several proteins, which normally exist in serum in an inactive form. The classic pathway is initiated by the complexing of antigen to its specific antibody, either IgG or IgM, and is the primary amplifier of the biologic effects of humoral immunity. Activation of the complement sequence leads to the consumption of complement components which, in turn, can lead to a decrease in their concentration. Thus, the determination of complement activity can indicate whether the complement system has been activated by an immunologic and/or pathogenic mechanism.
Increased complement levels are often associated with inflammatory conditions, trauma or acute illness such as myocardial infarction. Deficiencies of complement account for a small percentage of primary immunodeficiencies but depression of complement frequently co-exists with SLE and other disorders associated with an immunopathologic process. Thus, low levels of complement can be found in rheumatic diseases, glomerulonephritis, infectious diseases and in deficiency disorders.
The traditional method for determination of functional complement activity is the total hemolytic (CH50) assay. This assay measures the ability of the test sample to lyse 50% of a standardized suspension of sheep erythrocytes coated with anti-erythrocyte antibody. Both the classic activation and the terminal complement components are measured in this reaction. Total complement activity is usually abnormal if any component is defective.
Assessment of CH50 is useful in screening for genetic deficiencies in the complement system and in monitoring the progress of patients with immune complex disease.Principle of the Procedure
Total complement consists of a number of distinct components. When sheep erythrocytes are sensitized by antibody against sheep erythrocytes an antigen-antibody complex is formed. This complex, when exposed to the complement in human serum, will activate the components resulting in lysis of the erythrocytes and the release of hemoglobin. The degree of lysis is proportional to the concentration of total complement in the serum.
The Diamedix immunosimplicity (Is®) EZ Complement CH50 Test consists of test tubes that contain sheep erythrocytes sensitized with antibodies against sheep erythrocytes in 3 ml of standardized buffer solution.
The cell suspension has been adjusted so that approximately 5 µl of sample from an individual with normal complement levels will lyse approximately 50% of the cells. Cell lysis can be read on a standard spectrophotometer at 415 nm. Patient values are then compared to a Reference Serum with a known CH50 value.Please refer to complete Package Insert for greater detail about the test.
Features & Benefits
Formatted Procedure
