Enzyme-linked lectinsorbent assay measures N-acetyl-D-glucosamine in matrix of biofilm produced by Staphylococcus epidermidis.
An enzyme-linked lectinsorbent assay (ELLA) was developed for quantification of in situ biofilm produced by Staphylococcus epidermidis in polystyrene 96-well tissue culture plates with phosphatase-labeled wheat germ agglutinin (WGA) as a specific probe for the GlcNAcbeta-1,4n component of exocellular matrix material (EMM) that is responsible for intercellular adhesion and accumulation. The ELLA and the modified Christensen dye assay were used to test 13 laboratory strains of coagulase-negative staphylococci and 10 clinical isolates of S. epidermidis. Four biofilm-positive laboratory strains of S. epidermidis were positive by both tests, and six biofilm-negative strains were negative by both. One strain of S. hemolyticus was positive by the ELLA only. Two of the 10 clinical isolates of S. epidermidis were positive by both assays, two were negative by both, and the remaining were positive by the ELLA only. The ELLA was objective, reproducible, specific, sensitive, and useful for screening strains for their capacity to adhere to plastic, produce EMM, and form biofilm.
Thomas
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VL
Department of Microbiology, 7703 Floyd Curl Drive, The University of Texas Health Science Center at San Antonio, San Antonio, Texas 78284-7758, USA.
Sanford
B A
BA
Moreno
R
R
Ramsay
M A
MA
D000117:Acetylglucosamine / Q000032:analysis* / Q000096:biosynthesis*; D000469:Alkaline Phosphatase; D018441:Biofilms; D005069:Evaluation Studies as Topic; D006801:Humans; D037102:Lectins; D008722:Methods; D015203:Reproducibility of Results; D013212:Staphylococcus epidermidis / Q000302:isolation & purification / Q000378:metabolism*; D014909:Wheat Germ Agglutinins
D037102:Lectins; D014909:Wheat Germ Agglutinins; D000469:Alkaline Phosphatase; D000117:Acetylglucosamine