Reactivities of genus-specific monoclonal antibody EM-6E11 against Listeria species and serotypes of Listeria monocytogenes grown in nonselective and selective enrichment broth media.
Depending on the growth medium used for enrichment of bacterial cells prior to assay, the monoclonal antibody (MAb) EM-6E11 recognizing Listeria genus-specific epitope on 43 and 94 to 97 kDa cell-surface antigens (A. K. Bhunia and M. G. Johnson, Appl. Environ. Microbiol. 58:1924-1929, 1992) exhibited extensive variability in the detection of Listeria species. MAb EM-6E11 strongly detected live cells of all Listeria species and all serotypes of L. monocytogenes by ELISA when cells were grown in nonselective brain heart infusion (BHI) broth, in selective Listeria enrichment broth (LEB), or in Listeria repair broth (LRB). In contrast, EM-6E11 detected only four of the thirteen serotypes of L. monocytogenes (serotypes 1/2c, 3b, 4ab, and 7) when cells were grown in the UVM1 formulation of Listeria enrichment broth (UVM1) or Fraser broth (FRB). This MAb failed to react with live cells of four other Listeria species, including L. ivanovii, L. welshimeri, L. grayi, and L. murrayi cells grown in UVM1 or FRB. Heating of Listeria cells at 100 degrees C for 20 min, irrespective of the enrichment media used, led to large losses of MAb EM-6E11 reactivity in ELISA, suggesting that the specific cell-surface epitopes involved may not be heat stable. Our results confirm that MAb EM-6E11 is suitable for detection of live cells but not heat-killed cells of Listeria spp. and can be used in conjunction with an enrichment step in BHI, LEB, or LRB but not in UVM1 or FRB.
Department of Food Science, University of Arkansas Biotechnology Center, Fayetteville, USA. ramakris@comp.uark.edu
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D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000954:Antigens, Surface / Q000276:immunology; D003470:Culture Media; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D006358:Hot Temperature; D008087:Listeria / Q000145:classification / Q000254:growth & development* / Q000276:immunology*; D008089:Listeria monocytogenes / Q000145:classification / Q000254:growth & development* / Q000276:immunology*; D012703:Serotyping; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D003470:Culture Media; D000939:Epitopes