Abstract
A new immunodiffusion technique in agarose gel for the quantification of complement-fixing antibodies is described. The test involves the incorporation of antigen and complement in a primary agarose gel. Heat-inactivated serum samples are allowed to diffuse radially from wells overnight at 4°C. A secondary gel, containing antibody-coated sheep erythrocytes, is layered on top of the first gel and the system is incubated for 45 min at 37°C. Where complement is fixed, i.e., around wells with positive serum samples, zones of unlysed cells appear. There is a straight line relationship between zone areas so produced and log2 serum titres obtained with the conventional complement fixation test. The method appears to be applicable to a variety of antigens. It has been found suitable for bacterial and viral antigens. The test can also be reversed, thus allowing the quantification of diffusible antigens in a gel containing immune serum and complement. This paper describes in detail the use of this method as a diagnostic tool for the assay of complement-fixing antibodies to the type-specific antigens of influenza virus in paired human sera.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Haaheim R. Single-radial-complement-fixation: a new immunodiffusion technique. 2. Assay of the antibody response to the internal antigens (MP and NP) of influenza A virus in human sera after vaccination and infection. Dev Biol Stand. 1977 Jun 1;39:481–484. [PubMed] [Google Scholar]
- Schild G. C., Pereira M. S., Chakraverty P. Single-radial-hemolysis: a new method for the assay of antibody to influenza haemagglutinin. Applications for diagnosis and seroepidemiologic surveillance of influenza. Bull World Health Organ. 1975;52(1):43–50. [PMC free article] [PubMed] [Google Scholar]
- Skaug K., Orstavik I., Ulstrup J. C. Application of the passive haemolysis test for the determination of rubella virus antibodies. Acta Pathol Microbiol Scand B. 1975 Aug;83(4):367–372. doi: 10.1111/j.1699-0463.1975.tb00114.x. [DOI] [PubMed] [Google Scholar]
- Skehel J. J., Schild G. C. The polypeptide composition of influenza A viruses. Virology. 1971 May;44(2):396–408. doi: 10.1016/0042-6822(71)90270-4. [DOI] [PubMed] [Google Scholar]
- Vesikari T., Vaheri A. Rubella: a method for rapid diagnosis of a recent infection by demonstration of the IgM antibodies. Br Med J. 1968 Jan 27;1(5586):221–223. doi: 10.1136/bmj.1.5586.221. [DOI] [PMC free article] [PubMed] [Google Scholar]