Rapid rate-kinetic turbidometric assay for quantitation of viral complement-fixing antibodies. 1985

R E Fulton, and V L Dininno

A rapid rate-kinetic turbidometric assay for the quantitation of viral complement fixing antibodies has been developed, using adenovirus as a model. The procedure is based on the turbidometric quantitation of intact sheep erythrocytes and measures the rate of hemolysis (change in absorbance at 640 nm/min/), at maximum velocity, occurring in the presence of residual complement not fixed by the antigen-antibody reaction. Reagents were standardized and assays performed using a microprocessor-controlled spectrophotometer with kinetic assay capability and a thermoregulated cell compartment. Eleven sera were assayed for complement fixing antibodies both by the conventional microtiter technique and by the rapid turbidometric method described here. Good correlation (r = 0.89) was obtained between the two procedures. Unlike the conventional complement fixation test, the rate-kinetic turbidometric complement fixation assay was found to be tolerant of variation in complement and antigen concentration, endpoint titers were objectively quantitated and, once reagents had been standardized, results could be obtained within 45-60 min. The technique is potentially adaptable to large-scale automation.

UI MeSH Term Description Entries
D003167 Complement Activation The sequential activation of serum COMPLEMENT PROTEINS to create the COMPLEMENT MEMBRANE ATTACK COMPLEX. Factors initiating complement activation include ANTIGEN-ANTIBODY COMPLEXES, microbial ANTIGENS, or cell surface POLYSACCHARIDES. Activation, Complement,Activations, Complement,Complement Activations
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
D006168 Guinea Pigs A common name used for the genus Cavia. The most common species is Cavia porcellus which is the domesticated guinea pig used for pets and biomedical research. Cavia,Cavia porcellus,Guinea Pig,Pig, Guinea,Pigs, Guinea
D006461 Hemolysis The destruction of ERYTHROCYTES by many different causal agents such as antibodies, bacteria, chemicals, temperature, and changes in tonicity. Haemolysis,Extravascular Hemolysis,Intravascular Hemolysis,Extravascular Hemolyses,Haemolyses,Hemolyses, Extravascular,Hemolyses, Intravascular,Hemolysis, Extravascular,Hemolysis, Intravascular,Intravascular Hemolyses
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000258 Adenovirus Infections, Human Respiratory and conjunctival infections caused by 33 identified serotypes of human adenoviruses. Pharyngo-Conjunctival Fever,Human Adenovirus Infections,Infections, Human Adenovirus,Adenovirus Infection, Human,Fever, Pharyngo-Conjunctival,Human Adenovirus Infection,Infection, Human Adenovirus,Pharyngo Conjunctival Fever
D000260 Adenoviruses, Human Species of the genus MASTADENOVIRUS, causing a wide range of diseases in humans. Infections are mostly asymptomatic, but can be associated with diseases of the respiratory, ocular, and gastrointestinal systems. Serotypes (named with Arabic numbers) have been grouped into species designated Human adenovirus A-G. APC Viruses,APC Virus,Adenovirus, Human,Human Adenovirus,Human Adenoviruses
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D000914 Antibodies, Viral Immunoglobulins produced in response to VIRAL ANTIGENS. Viral Antibodies
D000956 Antigens, Viral Substances elaborated by viruses that have antigenic activity. Viral Antigen,Viral Antigens,Antigen, Viral

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