Serotyping involves the production of antibodies following the injection of a suitable mammal with the microorganism or a specific extract of the organism. If an organism is non-flagellated then serotyping will be based on the somatic antigens. If the organism is flagellated then serotyping may also be based on the flagella antigens. Three antigenic sites are recognized: somatic (O) (German "Ohne") or body, flagella (H) (German "Hauch") or motility, and K (German "Kapsel"), e.g., Escherichia coli O157:H7. The O antigens are comprised of the O polysaccharides that are on the surface and are heat stabile. The K and H antigens are heat labile. With whole bacterial cells, agglutination methods are used. With soluble antigens such as toxins, precipitin or gel diffusion assays are used.

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