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Screening and preliminary application of a DNA aptamer for rapid detection of Salmonella O8

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Abstract

We report on a rapid method for the detection of Salmonella O8. It does not require an enrichment step but rather uses an aptamer as a probe that was selected by system evolution of ligands by exponential enrichment (SELEX) assay. Firstly, aptamer against Salmonella O8 was selected from a 78 bp random DNA library that was prepared in-vitro. The binding ability of the aptamers to target bacterium was examined by aptamer-linked immobilized sorbent assay. A high affinity aptamer was successfully selected from the initial random DNA pool, and its secondary structure was also investigated. Next, this high affinity aptamer B10 was used to recognize Salmonella O8 via fluorescence microscopy. The selected aptamer has a high specificity and high affinity against its target. We believe that the resulting fluorescence in-situ labeling assay is a potentially useful alternative in rapid screening and detection of foodborne pathogens.

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Acknowledgements

The authors wish to thank the Science and Technology Research Project of general administration of quality supervision, inspection and quarantine of P. R. China (2010IK135, 201210127), the 12th Five-Year Key Programs (2012BAK17B10) and the Huangshan Mountain Young Scholar Fund of Hefei University of Technology (407–037025) for financial support.

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Authors and Affiliations

  1. School of Biotechnology and Food Engineering, Hefei University of Technology, Hefei, 230009, People’s Republic of China

    Guoqing Liu, Wei Chen, Yongkan Ye, Xiaojiao Yang & Yingqi Lian

  2. Anhui Entry-exit Inspection and Quarantine Bureau, Hefei, 230022, People’s Republic of China

    Xiaofeng Yu, Yi Yan & Kai Zong

  3. Jiangsu Entry-exit Inspection and Quarantine Bureau, Nanjing, 200002, People’s Republic of China

    Feng Xue

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  1. Guoqing Liu
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  2. Xiaofeng Yu
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  3. Feng Xue
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  4. Wei Chen
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  5. Yongkan Ye
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  6. Xiaojiao Yang
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  7. Yingqi Lian
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  8. Yi Yan
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  9. Kai Zong
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Correspondence to Wei Chen or Kai Zong.

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Liu, G., Yu, X., Xue, F. et al. Screening and preliminary application of a DNA aptamer for rapid detection of Salmonella O8. Microchim Acta 178, 237–244 (2012). https://doi.org/10.1007/s00604-012-0825-2

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  • DOI: https://doi.org/10.1007/s00604-012-0825-2

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