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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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Abstract

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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Authors and Affiliations

  1. Department of Microbiology, Immunology and Biochemistry, The University of Tennessee Health Science Center, Memphis, TN, 38163, USA

    Fereidoun Forghani

  2. KU Center for Food Safety, College of Veterinary Medicine, Konkuk University, Seoul, 05029, Korea

    Kun-Ho Seo

  3. Department of Food Science and Biotechnology, School of Bioconvergence Science and Technology, Kangwon National University, Chuncheon, Gangwon, 24341, Korea

    Shuai Wei, Youn-Seo Park, Byung-Jae Park & Deog-Hwan Oh

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  1. Shuai Wei
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  2. Fereidoun Forghani
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  4. Byung-Jae Park
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Correspondence to Deog-Hwan Oh.

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Wei, S., Forghani, F., Park, YS. et al. Dynabeads protein G antibody conjugates combined with modified brain heart infusion broth for the enrichment and separation of Bacillus cereus in artificially contaminated vegetables. Food Sci Biotechnol 25, 941–947 (2016). https://doi.org/10.1007/s10068-016-0154-z

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  • DOI: https://doi.org/10.1007/s10068-016-0154-z

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