Lectin-biotin assay for slime present inin situ biofilm produced byStaphylococcus epidermidis using transmission electron microscopy (TEM) |
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Authors: | Dr B A Sanford V L Thomas S J Mattingly M A Ramsay M M Miller |
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Institution: | (1) Department of Microbiology, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, 78284-7758 San Antonio, Texas, USA;(2) Department of Pathology, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, 78284-7758 San Antonio, Texas, USA |
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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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Keywords: | S epidermidis biofilm slime lectin marker |
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