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Establishment and preliminary application of oligonucleotide microarray assay for detection of food-borne toxigenic microorganisms

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Abstract

Rapid, high-throughput and accurate detection and identification of food-borne toxigenic microorganisms is crucial for food safety nowadays. An oligonucleotide microarray was designed and established and was applied to detect common food-borne toxigenic microorganisms in this study. PCR amplification of marker genes and 16S rRNA gene of 14 toxigenic bacteria and fungi using specific primers and oligo probes residing in these genes were employed and designed to fabricate the microarray. Optimization of hybridization conditions was implemented. The optimal conditions for hybridization were 51 °C for 30 min. Furthermore, the ratio of biotin labeled to unlabeled primer for PCR amplification was also optimized to enhance specific hybridization of the microarray. Specificity, sensitivity (710 CFU/mL), and reproductivity assessment confirmed the practicability of the microarray. Finally, this microarray was successfully applied to detect 6 common toxigenic microorganisms from 328 food samples. The established microarray may provide potential for rapid detection and identification of toxigenic microorganisms from foods.

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Acknowledgments

We thank the technicians of Zhuhai Entry-Exit Inspection and Quarantine Bureau and for their kind help with sampling.

Conflict of interest

Guiyun Cao has received research grant from the Water Quality Research Center of Zhuhai Water Group Co., LTD. Jianwang Feng, Xiaoyu Wang, Songnan Hu, Xiaoshan Kuang, Shiming Tang, Shuzhu You and Lideng Liu declare that they have no conflict of interest.

Compliance with Ethics Requirements

This article does not contain any studies with human or animal subjects.

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Authors and Affiliations

  1. Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, 519015, China

    Jiawang Feng, Xiaoyu Wang, Songnan Hu, Xiaoshan Kuang, Shiming Tang, Shuzhu You & Lideng Liu

  2. The Water Quality Research Center of Zhuhai Water Group Co., Ltd., Zhuhai, 519000, China

    Guiyun Cao

Authors
  1. Jiawang Feng
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  2. Xiaoyu Wang
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  3. Guiyun Cao
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  4. Songnan Hu
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  6. Shiming Tang
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  7. Shuzhu You
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  8. Lideng Liu
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Corresponding author

Correspondence to Jiawang Feng.

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Feng, J., Wang, X., Cao, G. et al. Establishment and preliminary application of oligonucleotide microarray assay for detection of food-borne toxigenic microorganisms. Eur Food Res Technol 236, 1073–1083 (2013). https://doi.org/10.1007/s00217-013-1951-8

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  • DOI: https://doi.org/10.1007/s00217-013-1951-8

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