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Serological Tests

Bacterial species and types can be identified by specific antigen antibody reactions. Antigens are substances that induce the production of antibodies in a foreign species. Bacteria and bacterial components serve as excellent antigens. The test includes  production of antibodies in an animal host and testing of the antiserum by either the agglutination or precipitation test.In the agglutination test, a drop of the culture of a particular bacterium is mixed on a slide with the anti-serum of an individual infected by it and examined under a microscope.

If clumping occurs, the test bacterium is considered to be the same- or closely related to the bacterium used as the antigen.

The same test can be performed by the tube dilution technique which permits, the assay of antibody titer. In this, a constant amount of the antigen is added to a  series of tubes containing serum dilutions. After mixing, the tubes are incubated at a particular temperature and the highest dilution of serum showing visible agglutination is determined. Diagnostic antisera are used to identify and group the more common bacteria.

Precipitation test is used to check the similarity of soluble antigens. An antiserum is tested by allowing it to diffuse through an agar gel towards a source of crude extract of bacteria. A sharp line of precipitate forms in the zone of antigen antibody interaction. This test is performed by placing about 1.5 per cent solution of agar into a Petri dish, after the agar has hardened, a well is bored in the centre to hold the antiserum,

Various antigens to be tested are placed in wells, around the central source of antiserum leaving about 5-15 mm space between the wells.

The antigens and antibodies diffuse through the agar and encounter each other. White lines precipitate appear at the zones of optimal con­centration between those antigens having mutual reactivity.

The way in which precipitin lines intersect can reveal whether the antigens in adjacent wells have any common determinant groups.Three classes of surface antigens have been extensively studied among various enteric bacteria. These are O antigens (lipopolysaccharides in the outer membrane), K antigens (capsular polysac­charides) and H antigens (flagellar proteins).

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