Clinical Microbiology: Open Access

Detection of infection with Actinomyces israelii by monoclonal antibodies 101-121 against Actinomyces israelii polysaccharide

6^th Clinical Microbiology Conference

October 20-22, 2016 Rome, Italy

Adnan Saeed, Pasciak Mariola, Szkudlarek Jerzy, Drab Marek and Gamian Andrzej

Polish Academy of Sciences, Poland

Posters & Accepted Abstracts: Clin Microbiol

Abstract:

Human actinomycosis, is rare a chronic, suppurative granulomatous infectious disease, has been recognized for over a century, caused by microorganisms of genus Actinomyces species. Presently, Actinomyces species comprises 42 species, 20 of them are relevant to human medicine. Actinomycosis is defined as a hard mass-type lesion with a specific histopathological structure. There are a large number of case reports of actinomycosis in the literature, but in most cases, diagnosis has been based solely on clinical and histopathological findings. The goal of the study was to determine the chemical composition of polysaccharide antigens extracted from A. israelii and generate monoclonal antibody reactive with the polysaccharide to understand their role in pathogenicity. Polysaccharides were extracted from dry bacterial cell mass by using trichloroacetic acid (TCA) and enzymes (DNase, RNase and protease). Further were purified by ion-exchange chromatography (DEAE Sephadex A25) and gel filtration (TOYOPEARL HW 55 s). Composition and structure of polysaccharide was determined by gas liquid chromatography-mass spectrometry GLC-MS. Monoclonal antibodies were generated by the hybridoma technique. The ELISA method was carried out for evaluating specificity of monoclonal antibody to the polysaccharide antigen. Then quantitative immunoprecipitation test has been performed. The experimental infection of mice with A. israelii was performed and investigated in histological study. Detection of the bacterial strain within the mouse tissues was done by immunohistochemical test. Scanning electron microscopy showed a variation of the branched shape of A. israelii with filamentous character of slime. Polysaccharide of A. israelii consists of glucose, galactose and mannosamine in the molar ratio 1, 5, 10 respectively. Two hybridomas 101 and 121 producing mAbs against polysaccharide antigen were IgM class. Quantitative microimmunoprecipitation test showed that monoclonal antibody precipitated polysaccharide antigen of A. israelii. Immunohistochemical test with monoclonal antibodies identified A. israelii infection in the liver mouse tissue. Monoclonal antibodies with polysaccharide used in immunohistochemical assays could serve as tools for diagnostic purposes in vitro approaches.

Biography :

Email: adnan.m14.m14@gmail.com