An ELISA for measuring serum antibodies to Actinobacillus actinomycetemcomitans

Abstract

An adaptation of the indirect enzyme‐linked immunosorbent assay (ELISA) is described for the determination of isotype specific antibodies directed to intact Actinobacillus actinomycetemcomitans and bacterial sonicates. Sera from nine individuals, with or without diagnosed periodontal disease, have been examined for IgG antibodies to the microorganism. The conditions for the assay have been studied and the ELISA has been compared to bacterial agglutination, indirect immunofluorescence or passive hemagglutination. A positive correlation is noted among the analyses in separating the sera into high, moderate and low antibody activities. However, based upon the highest dilution of the sera that gave positive reactions in the assays, the modified ELISA is 5–50 fold more sensitive than either the microagglutination, indirect immunofluorescence or passive hemagglutination techniques as performed. The ELISA is a sensitive assay that uses small quantities of biological materials to determine isotype specific antibody levels. This assay can provide a means for rapid screening of large numbers of biological samples for antibodies to intact microorganisms.

Multiserotype Enzyme-Linked Immunosorbent Assay as a Diagnostic Aid for Periodontitis in Large-Scale Studies

“…Serum IgG antibodies against A. actinomycetemcomitans and P. gingivalis were determined by an ELISA essentially as described earlier (6). As antigens we used mixtures of six strains of A. actinomycetemcomitans and three strains of P. gingivalis.…”

B and T Lymphocytes Are the Primary Sources of RANKL in the Bone Resorptive Lesion of Periodontal Disease

“…22 This periodontal bone resorption was abrogated by OPG-Fc local injection. 22 Accumulation of plasma cells and B cells in the disease lesion 11,12 and elevated IgG antibodies to periodontal bacteria in the serum 1,2,28 and gingival crevice fluid 40 are distinct features of periodontitis. However, the nature of B cells in the context of periodontal bone resorption has not been clearly elucidated until our previous study.…”

“…The method for detection of IgG response to oral bacteria has been previously published. 27,28 To detect anti-bacterial IgG levels in the gingival homogenates, the following formalin-fixed bacteria were used as antigen in ELISA: A. actinomycetemcomitans strain Y4 (American Type Culture Collection no. 43718), Fusobacterium nucleatum str.…”

Chemical and biological activities of a 64-kilodalton outer sheath protein from Treponema denticola strains

“…The pellet was resuspended to two-thirds the original volume with phosphate-buffered saline (PBS) and dialyzed against PBS at 4°C for 24 h, and the protein content was determined. Antibody titers to the 64-kDa outer sheath protein, using the 33% ammonium sulfate IgG-enriched fraction against formalinized whole cells of GM-1, were determined by an enzymelinked immunosorbent assay by the method of Ebersole et al (18). Antibody titers of 1:102,400 were achieved.…”