Abstract
Clostridium septicum produces a number of diseases in human and farm animals which, in most of the cases, are fatal without clinical intervention. Alpha toxin is an important agent and the unique lethal virulent factor produced by Clostridium septicum. This toxin is haemolytic, highly lethal and necrotizing activities but is being used as an antigen to develop animal vaccines. The aim of this study was to isolate the alpha toxin of Clostridium septicum and produce highly specific antibodies against it. In this work, we have developed a simple and efficient method for alpha toxin purification, based on electroelution that can be used as a time-saving method for purifying proteins. This technique avoids contamination by other proteins that could appear during other protein purification techniques such chromatography. The highly purified toxin was used to produce polyclonal antibodies. The specificity of the antibodies was tested by western blot and these antibodies can be applied to the quantitative determination of alpha toxin by slot blot.
Highlights
Clostridium is spore forming, Gram-positive and anaerobic
The protein profile analyses of samples revealed the presence of different proteins, all of them belonging to C. septicum
Three of them were identified as C. septicum alpha toxin: a high molecular weight protein and two proteins of about 49 kDa and 43 kDa
Summary
Clostridium is spore forming, Gram-positive and anaerobic ( some species are microaerophilic). Fourteen species that are clearly or potentially pathogenic produce biologically active proteins (toxins), which are responsible for their pathogenicity; some of them are fatal (Hatheway, 1990). Clostridium can affect the muscle and subcutaneous tissues of cattle, sheep, goats and other animal species (Rahman et al, 2009). Clostridium septicum is a pathogen which can cause various disease syndromes in animals and humans. Animal disease syndromes are slightly less understood than in humans (Tweten, 2001). C. septicum is the primary etiological agent of a traumatic clostridial myonecrosis, a rapidly fulminating and frequently fatal necrotic disease of the human musculature and was found to be a major cause of gas gangrene due to infection of
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