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KMID : 1007520160250030941
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Food Science and Biotechnology
2016 Volume.25 No. 3 p.941 ~ p.947
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Dynabeads Protein G Antibody Conjugates Combined with Modified Brain Heart Infusion Broth for the Enrichment and Separation of Bacillus cereus in Artificially Contaminated Vegetables
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Wei Shuai
Fereidoun Forghani
Park Youn-Seo
Park Byung-Jae
Seo Kun-Ho
Oh Deog-Hwan
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Abstract
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A modified brain heart infusion (MBHI) broth and a protocol of immunomagnetic separation (IMS) using antibody-coated Dynabeads® protein G were developed for the enrichment and separation of Bacillus cereus in artificially contaminated vegetable samples. The MBHI consisted of BHI and 0.34 g/L magnesium sulfate, 12.08 g/L sodium pyruvate, 1.82 g/L yeast extract, and polymyxin B. The amount of immunomagnetic beads (IMBs) and immunoreaction time were optimized. The capture efficiency was 58.32% with 0.4 mg IMBs when the immunoreaction time was 20 min. Capture of B. cereus by IMBs did not interfere with competing flora. Pre-enrichment IMS was validated with four B. cereus strains in artificially contaminated baby sprouts, bean sprouts, lettuce, and spinach at two levels (~0.1 and ~1 CFU/g). We were able to detect and isolate B. cereus in 40/40 samples of vegetables contaminated at 0.1 CFU/g with IMS after 6 h of enrichment in MBHI.
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KEYWORD
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Bacillus cereus, immunomagnetic separation, Dynabeads® protein G, enrichment
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