Difference between revisions of "Agglutination"
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| − | + | ==Introduction== | |
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Agglutination is the clumping together of particles, a reaction that can be used as an immunological test by exploiting the interaction between antibody and a particulate antigen. The antibodies that produce such a reaction are known as agglutinins. | Agglutination is the clumping together of particles, a reaction that can be used as an immunological test by exploiting the interaction between antibody and a particulate antigen. The antibodies that produce such a reaction are known as agglutinins. | ||
| − | + | ==Method== | |
The agglutination test is often carried out in round-bottomed test tubes, with doubling dilutions of the antiserum (i.e. 1:2, 1:4, 1:8...). The particulate antigen is then added and the mixture incubated at 37 degrees C. The last tube showing clear agglutination constitutes the end of the test. The dilution at which the test ends is known as the '''titre'''- the number of antibodies per unit volume of the serum. | The agglutination test is often carried out in round-bottomed test tubes, with doubling dilutions of the antiserum (i.e. 1:2, 1:4, 1:8...). The particulate antigen is then added and the mixture incubated at 37 degrees C. The last tube showing clear agglutination constitutes the end of the test. The dilution at which the test ends is known as the '''titre'''- the number of antibodies per unit volume of the serum. | ||
*An excess of antibody can inhibit agglutination reactions, again in much the same way as a precipitation reaction- this is known as the '''prozone effect'''. At high antibody concentrations, the number of epitopes is outnumbered by antibody binding sites. This means the antibodies bind to the antigen univalently, so are unable to cross-link antigen. This often happens in the first few tubes of the test, so the agglutination often only happens in those containing more dilute antiserum. | *An excess of antibody can inhibit agglutination reactions, again in much the same way as a precipitation reaction- this is known as the '''prozone effect'''. At high antibody concentrations, the number of epitopes is outnumbered by antibody binding sites. This means the antibodies bind to the antigen univalently, so are unable to cross-link antigen. This often happens in the first few tubes of the test, so the agglutination often only happens in those containing more dilute antiserum. | ||
| − | + | ==Applications== | |
| − | [[Image:Coombs test schematic.png|thumb|right|100px|Coombs test | + | [[Image:Coombs test schematic.png|thumb|right|100px|Coombs test. A Rad, '''WikiMedia Commons''' 2006]] |
*'''Widal test''': used to test for antibodies to certain bacteria in patient serum, e.g. salmonella | *'''Widal test''': used to test for antibodies to certain bacteria in patient serum, e.g. salmonella | ||
*'''Blood typing''': also known as '''haemagglutination''', the reaction can be used to determine blood type, i.e. an anti-A serum will agglutinate with type A red blood cells but not with B or O. | *'''Blood typing''': also known as '''haemagglutination''', the reaction can be used to determine blood type, i.e. an anti-A serum will agglutinate with type A red blood cells but not with B or O. | ||
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*Hormone assay by agglutination inhibition- red blood cells coated with a specific hormone produce agglutination with a hormone-specific antibody. The addition of free hormone (the test sample) will block the antigen-binding sites, preventing agglutination. By comparing the activity of the test sample against that of a known standard hormone, a quantitative result can be obtained. | *Hormone assay by agglutination inhibition- red blood cells coated with a specific hormone produce agglutination with a hormone-specific antibody. The addition of free hormone (the test sample) will block the antigen-binding sites, preventing agglutination. By comparing the activity of the test sample against that of a known standard hormone, a quantitative result can be obtained. | ||
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| + | {{review}} | ||
| + | <br><br> | ||
| + | {{Jim Bee 2007}} | ||
[[Category:Immunological Testing]] | [[Category:Immunological Testing]] | ||
Latest revision as of 16:48, 17 March 2012
Introduction
Agglutination is the clumping together of particles, a reaction that can be used as an immunological test by exploiting the interaction between antibody and a particulate antigen. The antibodies that produce such a reaction are known as agglutinins.
Method
The agglutination test is often carried out in round-bottomed test tubes, with doubling dilutions of the antiserum (i.e. 1:2, 1:4, 1:8...). The particulate antigen is then added and the mixture incubated at 37 degrees C. The last tube showing clear agglutination constitutes the end of the test. The dilution at which the test ends is known as the titre- the number of antibodies per unit volume of the serum.
- An excess of antibody can inhibit agglutination reactions, again in much the same way as a precipitation reaction- this is known as the prozone effect. At high antibody concentrations, the number of epitopes is outnumbered by antibody binding sites. This means the antibodies bind to the antigen univalently, so are unable to cross-link antigen. This often happens in the first few tubes of the test, so the agglutination often only happens in those containing more dilute antiserum.
Applications
- Widal test: used to test for antibodies to certain bacteria in patient serum, e.g. salmonella
- Blood typing: also known as haemagglutination, the reaction can be used to determine blood type, i.e. an anti-A serum will agglutinate with type A red blood cells but not with B or O.
- Virus diagnosis: some viruses, such as the myxoviruses, cause agglutination of the red blood cells. Antibodies inhibit this process and can be detected in a patient's serum for diagnosis.
- Coombs test: agglutination of red blood cells is an indication that certain antibodies are present on the cell surface, e.g. testing for Rh disease in newborn babies.
- Hormone assay by agglutination inhibition- red blood cells coated with a specific hormone produce agglutination with a hormone-specific antibody. The addition of free hormone (the test sample) will block the antigen-binding sites, preventing agglutination. By comparing the activity of the test sample against that of a known standard hormone, a quantitative result can be obtained.
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This article has been peer reviewed but is awaiting expert review. If you would like to help with this, please see more information about expert reviewing. |
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Originally funded by the RVC Jim Bee Award 2007 |