title: Purification and Evaluation of Polysaccharide Intercellular Adhesion (PIA) Antigen from Staphylococcus epidermidis.
abstract: The polysaccharide intercellular adhesin (PIA) confers major functional effects in biofilm formation, which bears an important role in the pathogenicity of Staphylococcus epidermidis. Following the identification of biofilm-forming strains by biochemical and molecular methods, isogenic strain was prepared and in vitro biofilm formation assay was performed consequently. By parallel analysis of both the PIA-positive and PIA-negative strains using size exclusion chromatography by Fast protein liquid chromatography (FPLC) method, the respective PIA was purified. Recovered PIA was examined using colorimetric and hemagglutination assays. Finally, the recovered PIA was analyzed using Fourier-transform infrared spectroscopy and proton nuclear magnetic resonance spectroscopy methods. By the parallel purification process and comparison of the obtained graphs from the FPLC detector, fractions near the void volume were determined as PIA. The colorimetric and hemagglutination assays were applied and the content of carbohydrates (hexose = 620 µg/ml, hexosamine = 5700 µg/ml and ketoses = 170 µg/ml) and hemagglutination titer (1:128) in recovered polysaccharide were determined. This study shows that PIA has a significant role in the biofilm formation in S. epidermidis strains. The recovered polysaccharide and its molecular weight were analyzed within the near void volume of the utilized column.
authors.affiliation: Department of Microbiology, Microbial Research Center, Pasteur Institute of Iran, Tehran, Iran.|Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Bahman
authors.initials: B
authors.lastname: Mirzaei
authors.affiliation: Department of Microbiology, Microbial Research Center, Pasteur Institute of Iran, Tehran, Iran.|Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Seyed Fazlollah
authors.initials: SF
authors.lastname: Moosavi
authors.affiliation: Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran.|Department of Mycobacteriology and pulmonary research, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Ryhane
authors.initials: R
authors.lastname: Babaei
authors.affiliation: Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran. d.siadat@gmail.com.|Department of Mycobacteriology and pulmonary research, Pasteur Institute of Iran, Tehran, Iran. d.siadat@gmail.com.
authors.forename: Seyed Davar
authors.initials: SD
authors.lastname: Siadat
authors.affiliation: Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran.|Department of Mycobacteriology and pulmonary research, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Farzam
authors.initials: F
authors.lastname: Vaziri
authors.affiliation: Laboratory of Medical Microbiology, Department of Medical Diagnostic Sciences, KU Leuven, U.Z. Gasthuisberg, Herestraat 49 CDG 8th floor, 3000, Leuven, Belgium.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Shahrooei
mesh_terms: D018829:Adhesins, Bacterial / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D018441:Biofilms; D011135:Polysaccharides, Bacterial / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D017550:Spectroscopy, Fourier Transform Infrared; D013212:Staphylococcus epidermidis / Q000737:chemistry* / Q000235:genetics / Q000502:physiology
chemical_list: D018829:Adhesins, Bacterial; D011135:Polysaccharides, Bacterial; C113579:polysaccharide intercellular adhesin