Hemagglutination assay

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Jump to: navigation, search The Haemagglutination Assay (HA) is a method of quantification for viruses or bacteria by hemagglutination. It is an easy, simple and rapid method which can be applied to large numbers of samples. Some viral families and many bacteria have envelope or surface proteins which are able to agglutinate (stick to) human or animal Red blood cells (RBC) and bind to N-acetylneuraminic acid. As each of the agglutinating molecule attaches to multiple RBCs, a lattice-structure will form. The haemagglutination assay of a virus, in contrast to other forms of virus quantification such as a plaque assay or 50% Tissue Culture Infective Dose, does not give any measure of viral infectivity, because no virus replication is required in this assay. The same may not be true when using HA for bacteria. The detailed conditions depend on the type of virus or bacteria being assayed since certain pH values and ionic strengths can impact the activity of the proteins of interest in a difficult to predict manner. Normally, a virus dilution (e.g. 2-fold from 1:4 to 1:4096) will be applied to an RBC dilution (e.g. 0.1% to 0.7% in steps of 0.2%) for approx. 30 min, often at 4C, otherwise viruses with neuraminidase activity will detach the virus from the RBCs. Then the lattice forming parts will be counted and the titre calculated. Virus concentration in virions per milliliter = 107 x HA titer.[1] For bacteria, depending on species, a bacterial dilution will be applied to an equal part RBC dilution and then incubated for 30 min to an hour at an optimal growth temperature before being observed.[2]

[edit] See also

Hemagglutination

[edit] References
1. ^ Donald HB, Isaacs A (1954). "Counts of influenza virus particles". J. Gen. Microbiol. 10 (3): 45764. PMID 13174769. 2. ^ X. Chen , et al. (2007). "The S-Layer Proteins of L. crispatus strain ZJ001 is responsible for competitive exclusion against E. coli O157:h7 and S. typhimurim". Int. J. Food Microbiology 115 (3): 307312. doi:10.1016/j.ijfoodmicro.2006.11.007. PMID 17289201.

Agglutination (biology)
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Agglutination is the clumping of particles. The word agglutination comes from the Latin agglutinare, meaning "to glue."[1] This occurs in biology in three main examples:
1. The clumping of cells such as bacteria or red blood cells in the presence of an antibody. The antibody or other molecule binds multiple particles and joins them, creating a large complex. 2. The coalescing of small particles that are suspended in a solution; these larger masses are then (usually) precipitated. 3. An allergic reaction type occurrence where cells become more compacted together to prevent foreign materials entering them. This is usually the result of an antigen in the vicinity of the cells.

Contents
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1 Agglutination in hematology o 1.1 Hemagglutination o 1.2 Leukoagglutination 2 Agglutination in microbiology 3 History of discoveries 4 See also 5 References

[edit] Agglutination in hematology

[edit] Hemagglutination

The 'bedside card' method of blood typing, in this case using a Serafol card. The result is blood group A positive.

Hemagglutination is when the particles involved are red blood cells. The agglutin is called hemagglutinin. In cross-matching, agglutination occurring when donor red blood cells and recipient's serum or plasma are incubated together indicates that the donor blood is incompatible for that particular recipient.
[edit] Leukoagglutination

Leukoagglutination is when the particles involved are white blood cells. An example is the PH-L form of phytohaemagglutinin.

[edit] Agglutination in microbiology

Agglutination is commonly used as a method of identifying specific bacterial antigens, and in turn, the identity of such bacteria. Because the clumping reaction occurs quickly and is easy to produce, agglutination is an important technique in diagnosis.

[edit] History of discoveries

Two bacteriologists, Herbert Edward Durham (-1945) and Max von Gruber (18531927), discovered specific agglutination in 1896. The clumping became known as Gruber-Durham

reaction. Gruber introduced the term agglutinin (from the Latin) for any substance that caused agglutination of cells. French physician Fernand Widal (18621929) put Gruber and Durham's discovery to practical use later in 1896, using the reaction as the basis for a test for typhoid fever. Widal found that blood serum from a typhoid carrier caused a culture of typhoid bacteria to clump, whereas serum from a typhoid-free person did not. This Widal test was the first example of serum diagnosis. Austrian physician Karl Landsteiner found another important practical application of the agglutination reaction in 1900. Landsteiner's agglutination tests and his discovery of ABO blood groups was the start of the science of blood transfusion and serology which had made transfusion possible and safe after a while.

[edit] See also

Macrophage Immune System Blocking antibody

[edit] References
1. ^ agglutination, Mosby's Medical, Nursing and Allied Health Dictionary, Fourth Edition,kMosby Year-Book, 1994, p. 49