Factors affecting assay of myoglobin by complement fixation or immunodiffusion. 1975

J E Olerud, and D L Clark

Quantitative micro-scale complement-fixation and semi-quantitative gel immunodiffusion assays for myoglobin have been evaluated experimentally and used to measure myoglobin in the sera of more than 1500 patients. We report certain problems, observations, and methodological improvements. Serum caused enhancement of precipitin lines in the immunodiffusion assay. In the complement-fixation assay, serum interfered both by enhancing or inhibiting complement fixation. Suggested modifications in the complement-fixation assay are: (a) routine threefold or greater dilution of serum with buffer and (b) use of serum-based rather than buffer-based calibration materials in preparing standard curves. Three cycles of freeze-thaw of serum or buffer decreased detectable myoglobin by 50-100%. We conclude that certain serum-related factors and freeze-thaw phenomena are important causes of error in immunological assays for myoglobin.

UI MeSH Term Description Entries
D008832 Microchemistry The development and use of techniques and equipment to study or perform chemical reactions, with small quantities of materials, frequently less than a milligram or a milliliter.
D009211 Myoglobin A conjugated protein which is the oxygen-transporting pigment of muscle. It is made up of one globin polypeptide chain and one heme group.
D003168 Complement Fixation Tests Serologic tests based on inactivation of complement by the antigen-antibody complex (stage 1). Binding of free complement can be visualized by addition of a second antigen-antibody system such as red cells and appropriate red cell antibody (hemolysin) requiring complement for its completion (stage 2). Failure of the red cells to lyse indicates that a specific antigen-antibody reaction has taken place in stage 1. If red cells lyse, free complement is present indicating no antigen-antibody reaction occurred in stage 1. Complement Absorption Test, Conglutinating,Conglutination Reaction,Conglutinating Complement Absorption Test,Complement Fixation Test,Conglutination Reactions,Fixation Test, Complement,Fixation Tests, Complement,Reaction, Conglutination,Reactions, Conglutination,Test, Complement Fixation,Tests, Complement Fixation
D005069 Evaluation Studies as Topic Works about studies that determine the effectiveness or value of processes, personnel, and equipment, or the material on conducting such studies. Critique,Evaluation Indexes,Evaluation Methodology,Evaluation Report,Evaluation Research,Methodology, Evaluation,Pre-Post Tests,Qualitative Evaluation,Quantitative Evaluation,Theoretical Effectiveness,Use-Effectiveness,Critiques,Effectiveness, Theoretical,Evaluation Methodologies,Evaluation Reports,Evaluation, Qualitative,Evaluation, Quantitative,Evaluations, Qualitative,Evaluations, Quantitative,Indexes, Evaluation,Methodologies, Evaluation,Pre Post Tests,Pre-Post Test,Qualitative Evaluations,Quantitative Evaluations,Report, Evaluation,Reports, Evaluation,Research, Evaluation,Test, Pre-Post,Tests, Pre-Post,Use Effectiveness
D005615 Freezing Liquids transforming into solids by the removal of heat. Melting
D005779 Immunodiffusion Technique involving the diffusion of antigen or antibody through a semisolid medium, usually agar or agarose gel, with the result being a precipitin reaction. Gel Diffusion Tests,Diffusion Test, Gel,Diffusion Tests, Gel,Gel Diffusion Test,Immunodiffusions,Test, Gel Diffusion,Tests, Gel Diffusion
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man

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