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Lectin-biotin assay for slime present inin situ biofilm produced byStaphylococcus epidermidis using transmission electron microscopy (TEM)

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Journal of Industrial Microbiology

Abstract

A lectin-biotin assay was developed for use in the specific detection of slime produced byStaphylococcus epidermidis RP62A and M187sp11 grown in a chemically defined medium. Mature biofilm was formed on polyvinylchloride (PVC) disks using a combined chemostat-modified Robbins device (MRD) model system. Specimens fixedin situ were: 1) stained with ruthenium red; 2) reacted overnight with biotin-labeled lectins (WGA, succinyl-WGA, Con A, or APA) followed by treatment with gold-labeled extravidin; or 3) reacted with antibodies againstS. epidermidis RP62A capsular polysaccharide/adhesin (PS/A) using an immunogold procedure. WGA and succinyl-WGA (S-WGA), which specifically bindN-acetylglucosamine, were shown by TEM to react only with slime, both cell-associated and exocellular. In contrast, Con A, APA and anti-PS/A reacted with the bacterial cell surface but did not react with slime. These results indicate the usefulness of WGA lectin as a specific marker for detection of the presence and distribution of slime matrix material inS. epidermidis biofilm.

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Authors and Affiliations

  1. Department of Microbiology, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, 78284-7758, San Antonio, Texas, USA

    B A Sanford, V L Thomas, S J Mattingly, M A Ramsay & M M Miller

  2. Department of Pathology, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, 78284-7758, San Antonio, Texas, USA

    B A Sanford, V L Thomas, S J Mattingly, M A Ramsay & M M Miller

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  1. B A Sanford
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Sanford, B.A., Thomas, V.L., Mattingly, S.J. et al. Lectin-biotin assay for slime present inin situ biofilm produced byStaphylococcus epidermidis using transmission electron microscopy (TEM). Journal of Industrial Microbiology 15, 156–161 (1995). https://doi.org/10.1007/BF01569820

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  • DOI: https://doi.org/10.1007/BF01569820

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