Journal of Dairy Science and Biotechnology

Korean Society of Dairy Science and Biotechnology (한국낙농식품응용생물학회)
권호 표지

Rapid, Sensitive, and Specific Detection of Salmonella Enteritidis in Contaminated Dairy Foods using Quantum Dot Biolabeling Coupled with Immunomagnetic Separation

  • Kim, Hong-Seok (KU Center for Food Safety and College of Veterinary Medicine, Konkuk University) ;
  • Chon, Jung-Whan (KU Center for Food Safety and College of Veterinary Medicine, Konkuk University) ;
  • Kim, Hyunsook (Dept. of Food & Nutrition, College of Human Ecology, Hanyang University) ;
  • Kim, Dong-Hyeon (KU Center for Food Safety and College of Veterinary Medicine, Konkuk University) ;
  • Yim, Jin-Hyuk (KU Center for Food Safety and College of Veterinary Medicine, Konkuk University) ;
  • Song, Kwang-Young (KU Center for Food Safety and College of Veterinary Medicine, Konkuk University) ;
  • Kang, Il-Byung (KU Center for Food Safety and College of Veterinary Medicine, Konkuk University) ;
  • Kim, Young-Ji (KU Center for Food Safety and College of Veterinary Medicine, Konkuk University) ;
  • Lee, Soo-Kyung (KU Center for Food Safety and College of Veterinary Medicine, Konkuk University) ;
  • Seo, Kun-Ho (KU Center for Food Safety and College of Veterinary Medicine, Konkuk University)
  • Received : 2015.11.13
  • Accepted : 2015.12.19
  • Published : 2015.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

Colloidal semiconductor CdSe-ZnS core-shell nanocrystal quantum dot (Qdot) are luminescent inorganic fluorophores that show potential to overcome some of the functional limitations encountered with organic dyes in fluorescence labeling applications. Salmonella Enteritidis has emerged as a major cause of human salmonellosis worldwide since the 1980s. A rapid, specific, and sensitive method for the detection of Salmonella Enteritidis was developed using Qdot as a fluorescence marker coupled with immunomagnetic separation. Magnetic beads coated with anti-Salmonella Enteritidis antibodies were employed to selectively capture the target bacteria, and biotin-conjugated anti-Salmonella antibodies were added to form sandwich immune complexes. After magnetic separation, the immune complexes were labeled with Qdot via biotin-streptavidin conjugation, and fluorescence measurement was carried out using a fluorescence measurement system. The detection limit of the Qdot method was a Salmonella Enteritidis concentration of $10^3$ colony-forming units (CFU)/mL, whereas the conventional fluorescein isothiocyanate-based method required over $10^5CFU/mL$. The total detection time was within 2 h. In addition to the potential for general nanotechnology development, these results suggest a new rapid detection method of various pathogenic bacteria from a complex food matrix.

Keywords

References

  1. FAO/WHO (The Food and Agriculture Organization of the United Nations (FAO) and the World Health Organization). 2004. Joint FAO/WHO workshop on Enterobacter sakazakii and other microorganisms in powdered infant formula. WHO, Geneva, 2 to 5 February 2004.
  2. Gerion, D., Pinaud, F., Williams, S. C., Parak, W. J., Zanchet, D., Weiss, S. and Alivisatos, A. P. 2001. Synthesis and properties of biocompatible water soluble silica-coated CdSe/ZnS semiconductor Quantum Dots. J. Phys. Chem. B 105:8861-8871. https://doi.org/10.1021/jp0105488
  3. Howarth, M., Takao, K., Hayashi, Y. and Ting, A. A, 2005. Targeting Quantum Dots to surface proteins in living cells with biotin ligase. PNAS 102:7583-7588. https://doi.org/10.1073/pnas.0503125102
  4. Kaul, Z., Yaguchi, T., Kaul, S. C., Hirano, T., Wadhwa, R. and Taira, K. 2003. Mortalin imaging in normal and cancer cells with quantum dot immunoconjugates. Cell Research 13:503-507. https://doi.org/10.1038/sj.cr.7290194
  5. Kloepfer, J. A., Mielke, R. E., Wong, M. S., Nealson, K. H., Stucky, G. and Nadeau, J. L. 2003. Quantun Dots as strain- and metabolism-specific microbiological labels, Appl. Environ. Microbiol. 69:4205-4213. https://doi.org/10.1128/AEM.69.7.4205-4213.2003
  6. Lee, L.Y., Ong, S. L., Hu, J. Y., Ng, W. J., Feng, Y., Tan, X. and Wong, S. W. 2004. Use of semiconductor quantum dots for protostable immunofluorescence labeling of Cryptosporidium parvum. Appl. Environ. Microbiol. 70:5732-5736. https://doi.org/10.1128/AEM.70.10.5732-5736.2004
  7. Ness, J. M., Akhtar, R. S., Latham, C. B. and Roth, K. A. 2003. Combined tyramide signal amplification and quantum sensitive and photostable immunofluorescence detect. J. Histochem. Cytochem. 51:981-987. https://doi.org/10.1177/002215540305100801
  8. Seo, K. H., Brackett, R. E., Frank, J. F. and Hilliard, S. 1998. Immunomagnetic separation and flow cytometry for rapid detection of Escherichia coli O157:H7. J Food Prot. 61:812-816. https://doi.org/10.4315/0362-028X-61.7.812
  9. Su, X. L. and Li, Y. 2004. Quantum dot biolabeling coupled with immunomagnetic separation for detection of Escherichia coli O157:H7. Anal Chem. 76:4806-4810. https://doi.org/10.1021/ac049442+
  10. Varshney, M., Yang, L., Su, X. and Li, Y. 2005. Magnetic nanoparticle-antibody conjugated for the separation of Escherichia coli O157:H7 in ground beef. J. Food Prot. 68:1804-1811. https://doi.org/10.4315/0362-028X-68.9.1804
  11. Wang, B. B., Wang, Q., Jin, Y. G., Ma, M. H. and Cai, Z. X. 2015. Two-color quantum dots-based fluorescence resonance energy transfer for rapid and sensitive detection of Salmonella on eggshells. J. Photochem. Photobio. A: Chem. 299:131-137. https://doi.org/10.1016/j.jphotochem.2014.10.020
  12. Wang, D., He, E. J., Rosenzweig, N. and Rosenzweig, Z. 2004. Superparamagnetic $Fe_2O_3$ beads - CdSe/ZnS Quantum Dots core shell nanocomposite particles for cell separation. Nano Letters 4:409-413. https://doi.org/10.1021/nl035010n
  13. Wang, L., Li, Y. and Mustapha, A. 2007. Rapid and simultaneous quantitation of Escherichia coli O157:H7, Salmonella, and Shigella in ground beef by multiplex real-time PCR and Immunomagenetic separation. J. Food Prot. 70:1366-1372. https://doi.org/10.4315/0362-028X-70.6.1366
  14. Yang, L. and Li, Y. 2005. Quantum dots as fluorescent labels for quantitative detection of Salmonella typhimurium in chicken carcass wash water. J. Food Prot. 68:1241-1245. https://doi.org/10.4315/0362-028X-68.6.1241
  15. Yang, L. and Li, Y. 2006. Simultaneous detection of Escherichia coli O157:H7 and Salmonella Typhimurium using quantum dots as fluorescence labes. Analyst. 131:394-401. https://doi.org/10.1039/B510888H
  16. Zhu, L., Ang, S. and Liu, W.T. 2004. Quantum dots as a novel immunofluorescent detection system for Cryptosporidium parvum and Giardia lamblia. Appl. Environ. Microbiol. 70:597-598. https://doi.org/10.1128/AEM.70.1.597-598.2004