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Enzyme-linked Immunosorbent Assay Strip Sensor for Rapid Detection of Staphylococcus aureus

Staphylococcus aureus 신속 검출을 위한 효소면역측정 스트립 센서

  • Park, So Jung (Department of Chemical Engineering, Hankyong National University) ;
  • Kim, Young-Kee (Department of Chemical Engineering, Hankyong National University)
  • 박소정 (한경대학교 화학공학부) ;
  • 김영기 (한경대학교 화학공학부)
  • Received : 2011.07.29
  • Accepted : 2011.09.21
  • Published : 2011.10.10

Abstract

In this study, an established enzyme-linked immunosorbent assay and immuno-chromatography technique are combined to fabricate an immuno-strip sensor for the detection of S. aureus. The immuno-strip is manufactured by using four different functional membranes. The capture antibody is immobilized on the nitrocellulose membrane due to the high affinity and the capillary action through porous membranes induces a flow of sample. A colorimetric signal is appeared according to the enzyme reaction and is analyzed by the digital camera (qualitative analysis) and home-made image analysis software (quantitative analysis). Under the optimal conditions, samples with S. aureus in the range of $2.7{\times}10^4{\sim}2.7{\times}10^7CFU/mL$ can be detected by the colorimetric method within 30 min.

본 연구에서는, enzyme-linked immunosorbent assay (ELISA)와 면역크로마토그래픽 기법을 결합하여 Staphylococcus aureus 검출을 위한 면역스트립을 제작하였다. 면역스트립은 4개의 서로 다른 기능을 가진 멤브레인을 이용하여 만들어졌다. 니트로셀룰로오스 멤브레인은 항체와의 결합력이 높기 때문에, 포획항체를 고정화하였고, 다공성 멤브레인들을 통하여 모세관 현상으로 인해 시료흐름을 유도하였다. 효소반응에 의해 생성된 발색신호는 디지털카메라와 자체 제작된 소프트웨어를 이용하여 정성, 정량분석 하였다. 최적의 분석조건 하에서 30 min 이내에 $2.7{\times}10^4{\sim}2.7{\times}10^7CFU/mL$ 범위의 S. aureus 농도를 측정할 수 있었다.

Keywords

References

  1. H. J. Lee, Korean Journal of Veterinary Public Health, 32, 81 (2008).
  2. J.-M. Chang, and T. J. Fang, Food Microbiology, 24, 745 (2007). https://doi.org/10.1016/j.fm.2007.03.005
  3. L. Yang and R. Bashir, Biotechnology Advances, 26, 135 (2008). https://doi.org/10.1016/j.biotechadv.2007.10.003
  4. S. G. Park, Y. O. Hwang, J. H. Jung, and K. M. Lee, J. Food Hyg. Saf., 16, 159 (2001).
  5. J.-H. Lee, K.-Y. Song, J.-Y. Hyeon, I.-G. Hwang, H.-S. Kwak, J.-A Han, Y.-H. Chung, and K.-H Seo, Korean J. Food Sci. Ani. Resour., 30, 410 (2010). https://doi.org/10.5851/kosfa.2010.30.3.410
  6. J. Y. Moon, E. J. Lee, and Y. B. Kim, Journal of Bacteriology and Virology, 34, 91 (2004).
  7. V. L. M. Rall, F. P. Vieira, R. Rall, R. L. Vieitis, Fernandes Jr., J. M. G. Candeias, K. F. G. Cardoso, and J. P. Araujo Jr., Veterinary Microbiology, 132, 408 (2008). https://doi.org/10.1016/j.vetmic.2008.05.011
  8. S.-H. Huang, Sensors and Actuators B, 127, 335 (2007). https://doi.org/10.1016/j.snb.2007.04.027
  9. E. C. Alocilja and S. M. Radke, Biosensors and Bioelectronics, 18, 841 (2003). https://doi.org/10.1016/S0956-5663(03)00009-5
  10. O. Lazcka, F. J. D. Campo, and F. X. Munoz, Biosensors and Bioelectronics, 22, 1205 (2007). https://doi.org/10.1016/j.bios.2006.06.036
  11. A. D. Taylor, J. Ladd, Q. Yu, S. C. J. Homola, and S. Jiang, Biosensors and Bioelectronics, 22, 752 (2006). https://doi.org/10.1016/j.bios.2006.03.012
  12. P. D. Skottrup, M. Nicolaisen, and A. F. Justesen, Biosensors and Bioelectronics, 24, 339 (2008). https://doi.org/10.1016/j.bios.2008.06.045
  13. S.-H. Paek, C.-W. Lee, C.-S. Lee, and S.-H. Yook, Korean J. Biotechnol. Bioeng., 11, 42 (1996).
  14. S. H. Kim, W. S. Jung, and S.-H. Paek, Korean J. Biotechnol. Bioeng., 18, 190 (2003).
  15. S. Ko and S. A. Grant, Biosensors and Bioelectronics, 21, 1283 (2006). https://doi.org/10.1016/j.bios.2005.05.017
  16. J.-K. Park, The Magazine of the Institute of Electronics Engineers of Korea, 28, 56 (2001).
  17. J. H. Lee, S. S. Yang, B. W. Kim, S. J. Sim, H. Chae, and H. C. Yoon, Colloids and Surfaces A: Physicochem. Eng. Aspects, 313-314, 509 (2008). https://doi.org/10.1016/j.colsurfa.2007.04.143
  18. D. Ivnitski, I. Abdel-Hamid, P. Atanasov, and E. Wilkins, Biosensor and Bioelectronics, 14, 599 (1999). https://doi.org/10.1016/S0956-5663(99)00039-1
  19. C.-H Chang and S.-J. Kim, Korean J. Vet. Res., 29, 503 (1989).
  20. C.-H. Yoon, J.-H. Cho, H.-I. Oh, M.-J. Kim, C.-W. Lee, J.-W. Choi, and S.-H. Peak, Biosensors and Bioelectronics, 19, 289 (2003). https://doi.org/10.1016/S0956-5663(03)00207-0
  21. K. Inoue, P. Ferrante, Y. Hirano, T. Yasukawa, H. Shiku, and T. Matsue, Talanta, 73, 886 (2007). https://doi.org/10.1016/j.talanta.2007.05.008
  22. J. A. Ho, H.-W. Hsu, and M.-R. Huang, Analytical Biochemistry, 330, 342 (2004). https://doi.org/10.1016/j.ab.2004.03.038
  23. M. Varshney, Y. Li, B. Srinivasan, and S. Tung, Sensors and Actuators B, 128, 99 (2007). https://doi.org/10.1016/j.snb.2007.03.045
  24. S.-M. Han, J.-H. Cho, I.-H. Cho, E.-H. Paek, H.-B. Oh, B.-S. Kim, C. Ryu, K. Lee, Y.-K. Kim, and S.-H. Paek, Analytica Chemica Acta, 587, 1 (2007). https://doi.org/10.1016/j.aca.2007.01.028
  25. C. Fernandez-Sanchez, C. J. Mcneil, K. Rawson, O. Nilsson, H. Y. Leung, and V. Gnanapragasam, Journal of Immunological Methods, 307, 1 (2005). https://doi.org/10.1016/j.jim.2005.08.014
  26. J.-G. Choi, W.-B. Shin, J.-H. Je, J.-Y. Kim, K.-H. Lee, M.-G. Kim, S.-D. Ha, K.-S. Kim, K.-Y. Kim, C.-H. Kim, and D.-H. Chung, Korean J. Food Sci. Technol., 39, 299 (2007).