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Purification, Characteriztion and Immunodetection of Bacillus Cereus Toxins using Chicken Antibodies (Ig Y)

Purushothama, B.R. (2011) Purification, Characteriztion and Immunodetection of Bacillus Cereus Toxins using Chicken Antibodies (Ig Y). [Student Project Report]

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Abstract

This Dissertation / Report is the outcome of investigation carried out by the creator(s) / author(s) at the department/division of Central Food Technological Research Institute (CFTRI), Mysore mentioned below in this page.

Item Type: Student Project Report
Additional Information: The genus Bacillus is a large and diverse group of bacteria belonging to the family Bacillaceae, Phylum Firmicutes. The species in this genus are aerobic or facultatively anaerobic, endospore forming, rod shaped gram positive bacteria widely distributed in nature, especially in the soil. B. cereus group known as the causative agents of food-borne illness belongs to Group I of the genus Bacillus. A variety of media have been used for the detection of these bacteria. However, the toxins elaborated in to the food during contamination of food can be distinguished only after reported food poisoning outbreaks. The detection process involves sample preparation, isolation of toxin followed by detection by HPLC or GC. Hence a rapid, sensitive, robust and specific method for the detection of these toxins in foods needs to be addressed. With these in view, studies were carried out on the production, purification and immunodetection of the toxins elaborated by the genus Bacillus cereus. The isolate chosen was a standard isolate procured from IMTECH, Chandigarh. The BHI broth grown culture elaborated two kinds of toxins, haemolytic toxin and cytotoxin. These were purified by size exclusion chromatography and characterized by HPLC and SDS-PAGE. SDS-PAGE analysis of the purified fraction pool II showed 3 bands with haemolytic toxin corresponding to B, L1 and L2 components. The Fraction pool I showed single band indicating that the toxins were electrophoretically pure. The molecular weight were ∼ 46KDa and ∼ 34 KDa for Haemolytic toxin and cytotoxin K respectively. The poultry were immunized with the purified toxins and competitive sandwich ELISA was carried out.
Uncontrolled Keywords: Bacillus cereus, food poisoning, Microbial toxins
Subjects: 500 Natural Sciences and Mathematics > 07 Life Sciences > 04 Microbiology > 02 Bacteriology
Divisions: Fermentation Technology and Bioengineering
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 31 May 2011 04:44
Last Modified: 28 Dec 2011 10:23
URI: http://ir.cftri.com/id/eprint/10041

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