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标题：Immunogold Labelling of Porphyromonas gingivalis in Pure Culture and in Apical Border Plaque
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作者：P. Barber ; A. Boyd ; H. N. Newman 等
期刊名称：Microbial Ecology in Health and Disease
印刷版ISSN：1651-2235
出版年度：1990
卷号：3
期号：4
DOI：10.3402/mehd.v3i4.7537
语种：English
出版社：Microbial Ecology in Health and Disease
摘要：In this study we have attempted to develop methodology to enable identification of periodontitis-related organisms in situ using a post-embedding immunogold labelling technique. Pure cultures of Porphyromonas gingivalis W50 strain and specimens of plaque were fixed in 0.5 per cent glutaraldehyde in 0.1 M sodium cacodylate buffer and subsequently embedded in LR white acrylic resin. This procedure resulted in good ultrastructural preservation of the tissue and good retention of antigenicity. Fluorescence microscopy of a range of organisms and immunogold labelling of ultrathin sections were then employed using a high titre (1:5 x 105 by ELISA) murine polyclonal antiserum to P. gingivalis W50. Analysis of specificity by fluorescence microscopy using 46 different bacterial species demonstrated minimal cross-reactivity of the test antiserum, with only Bacteroides corporis showing an unexplained cross-reaction. At the ultrastructural level, consistent labelling of the pure culture specimens was achieved and labelling in plaque was confined to selected Gram-negative coccoid cells, or to associated areas of matrix and polymorphonuclear cytoplasm. These results indicate that post-embedding immunogold labelling may be an appropriate technique for the identification of bacterial strains within a mixed flora such as that of dental plaque.Keywords: Immunogold; Gold probes; Dental plaque; Porphyromonas gingivalis; Chronic periodontitis.