exhibits the serological relationships of Listeria spp. Most L. monocytogenes isolates obtained from patients and the environment are type 1 or 4. More than 90% of L. monocytogenes isolates can be serotyped with commercially available sera. All nonpathogenic species, except L. welshimeri, share one or more somatic antigens with L. monocytogenes. Serotyping alone without thorough characterization, therefore, is not adequate for identification of L. monocytogenes.
Serology is useful when epidemiological considerations are crucial. Use a TSBye culture to inoculate Tryptose broth. Incubate for 24 h at 35° C, at which temperature flagella (H) antigen expression is reduced. Transfer to Tryptose agar slants and incubate for 24 h at 35° C. Wash both slants in a total of 3 ml Difco fluorescent antibody (FA) buffer and transfer to a sterile 16 x 125-mm screw-cap tube. Heat in a water bath at 80°C for 1 h. Sediment cells by centrifugation at 1600 g for 30 min. Remove 2.2-2.3 ml of supernatant fluid and resuspend the pellet in the remainder of buffer. Follow manufacturer's recommendations for sera dilution and agglutination procedure. If flagella (H) or somatic (O) sub-factor serotyping is desired, see Chapter 11.
exhibits the serological relationships of Listeria spp. Most L. monocytogenes isolates obtained from patients and the environment are type 1 or 4. More than 90% of L. monocytogenes isolates can be serotyped with commercially available sera. All nonpathogenic species, except L. welshimeri, share one or more somatic antigens with L. monocytogenes. Serotyping alone without thorough characterization, therefore, is not adequate for identification of L. monocytogenes.
Serology is useful when epidemiological considerations are crucial. Use a TSBye culture to inoculate Tryptose broth. Incubate for 24 h at 35° C, at which temperature flagella (H) antigen expression is reduced. Transfer to Tryptose agar slants and incubate for 24 h at 35° C. Wash both slants in a total of 3 ml Difco fluorescent antibody (FA) buffer and transfer to a sterile 16 x 125-mm screw-cap tube. Heat in a water bath at 80°C for 1 h. Sediment cells by centrifugation at 1600 g for 30 min. Remove 2.2-2.3 ml of supernatant fluid and resuspend the pellet in the remainder of buffer. Follow manufacturer's recommendations for sera dilution and agglutination procedure. If flagella (H) or somatic (O) sub-factor serotyping is desired, see Chapter 11.
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