Differentiation of coagulase-positive and coagulase-negative staphylococci by lectins and plant agglutinins

Abstract

MATERIALS AND METHODS Reagents. All chemicals, salts, and sugars were of the highest grade available. L. polyphemus (crude), T. vulgaris (pure), and other routinely used lectins were purchased from E-Y Laboratory (San Mateo, Calif.). M. indica extract was prepared from the dried seeds of commercially purchased mangoes. The seeds were pulverized in a micromill (Chemical Rubber Co., Columbus, Ohio) and extracted for 2 h at room temperature in 10 times their weight of phosphate-buffered saline (PBS: 0.05 M potassium phosphate, 0.15 M 547

Fluorescein Isothiocyanate-Labeled Lectin Analysis of the Surface of the Nitrogen-Fixing Bacterium Azospirillum brasilense by Flow Cytometry

“…For FITC-Con A, the buffer solution was 5 mM Tris hydrochloride (pH 7.0) with 15 mM NaCl, 1 mM CaCl2, and 1 mM MnCl2. For other FITC-lectins (SBA, WGA, LTA, GS I, and GS II), the buffer was PBS (pH 7.45), but for FITC-GS I and FITC-GS II, 5 mM CaCl2 was added to the buffer. The binding of FITC-lectins to the washed bacteria was accomplished by the incubation of 5 x 109 cells of A. brasilense with 50 ,ug of FITC-lectin in 0.5 ml of buffer solution.…”

“…Agglutination. Cells of A. brasilense were grown for 24 h under N2-fixing or NH4+-assimilating conditions and examined for lectin-mediated agglutination according to the published procedure (5). Cells were washed in a buffer solution containing 0.85% NaCl and 10 mM Bis-Tris hydrochloride (pH 7.4).…”

“…The role of bacterial cell surface recognition in the colonization of host tissue has been well documented for animal (31) and plant (11) systems. Analysis of the cell surface by using lectins has yielded useful information for the characterization of sugars in bacterial capsules and cell walls (5,9,28,32). Lectin-cell interactions may be detected by agglutination; however, quantitation of the degree of agglutination is not easily achieved.…”

Significance of Staphylocoagulase in Food Microbiology: A Review

“…Most clinical a n d food laboratories utilize the tube coagulase test as the most efficient a n d reliable method for distinguishing S. aureus from other Micrococcaceae (AOAC 1975;STSM 1965;Kloos and Smith 1980;Baird-Parker et al 1976;Davidson et al 1982). Several investigators (Sperber a n d Tatini 1975; a n d Rayman et al 1975) have determined that the coagulase test is a valid means of identifying s. aureus, provided t h a t only a firm clot that does not move when the tube is tipped, is considered a positive reaction.…”

Lectin typing of five medically important Candida species

“…Lectins are proteins of non-immune origin that bind specifically to carbohydrates. 7 The selectivity of lectins for microbial-surface sugars has been demonstrated for bacteria of the genera Staphylococcus, 8,9,10 Neisseria, 11,12,13 Haemophilus, 14 Campylobacter 15 and Streptococcus 16,17 as well as for protozoa 18 and yeasts of the genus Candida. 19,20,21,22 In the present study we evaluated our results about the use of lectins for the typing of clinical Candida isolates.…”