한국어병학회지 (Journal of fish pathology)

  • 제20권3호
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  • Pages.307-313
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  • 2007
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  • 1226-0819(pISSN)
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  • 2233-5412(eISSN)
한국어병학회 (The Korean Society of Fish Pathology)
권호 표지

Piezoelectric immunosensor for the detection of Edwardsiellosis

  • Hong, Sung-Rok (Faculty of Marine Bioscience and Technology, Department of Chemistry, Kangnung National University) ;
  • Choi, Suk-Jung (Faculty of Marine Bioscience and Technology, Department of Chemistry, Kangnung National University) ;
  • Jeong, Hyun-Do (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Hong, Su-Hee (Faculty of Marine Bioscience and Technology, Department of Chemistry, Kangnung National University)
  • 발행 : 20071200
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초록

In this study, a QCM biosensor was made to detect Edwardsiella tarda (E. tarda) using a specific antibody. A 9 MHz AT-cut piezoelectric wafer layered with two gold electrodes of 5mm diameter had a reproducibility of 0.1 Hz in frequency response and was used as the transducer of the QCM biosensor. Self assembled layer (SAM) was conformed on a quartz crystal by treating with 3-mer-captopropionic acid (MPA) and activated with N-ethyl-N'-(3-dimethyl-aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS). The resulting NHS group was further converted to hydrazide by the reaction with hydrazine. Aldehyde group was introduced into the carbohydrate moiety of anti-E. tarda antibody by the reaction with periodic acid and was used to immobilise the antibody through the reaction with hydrazide group on the electrode surface. A baseline was established in the presence of phosphate-buffered saline (PBS) and a resonant frequency (F1) was measured. Sample was added to the sensor surface and second resonant frequency (F2) was measured after unbound substances were washed out with PBS several times. Finally, the frequency shift (ΔF) representing the mass change was calculated by subtracting F2 from F1. After adding the oxidized anti-E. tarda antibody to the electrode surface containing hydrazide group, frequency shift of 288.811.4 Hz (mean S.E) was observed, thus proving that considerable amount of antibody was immobilized. In the immunoassay test, the frequency shift of 1877.75 Hz, 580.67 Hz, 221.39 Hz, 7.671.83 Hz (mean S.E) were observed at doses of 1000, 500, 100, 50 g of bacterial cells, respectively. It was also demonstrated that the prepared sensor chip was stable enough to withstand repeated surface regeneration with 0.2 M Tris-glycine and 1 % DMSO, pH 2.3 more than ten times.

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참고문헌

  1. Absolom D., R., Zingg, W. and Neumann, A. W.: Protein adsorption to polymer particles: role of surface properties. J. Biomed. Mater. Res. 21: 161-171, 1987 https://doi.org/10.1002/jbm.820210202
  2. Amandi, A., S.F. Hiu, J. S. Rohovec, and J. L. Fryer.: Isolation and characterization of Edwardsiella tarda from fall chinook salmon (Oncorhvnchus tshawytscha). Applied Environ. Microbiol. 43: 13801384, 1982
  3. Attili, B. S. and Suleiman, A. A.: A piezoelectric immunosensor for the detection of cortisol. Anal. Lett. 28: 2149-2159, 1995 https://doi.org/10.1080/00032719508000035
  4. Boveniser, J. S., Jacobs, M. B., Guilbault, G. G. and O'Sullivan, C. K.: The detection of Pseudomonas aeruginosa using the quartz crystal microbalance. Anal. Lett., 31: 1287-1295, 1998 https://doi.org/10.1080/00032719808002866
  5. Bunde, R. L., Jarvi, E. J. and Rosentreter, J. J.: Piezoelectric quartz crystal biosensors. Talanta, 46, pp. 1223-1236, 1998 https://doi.org/10.1016/S0039-9140(97)00392-5
  6. Byers, H. K., Cipriano, R. C. Gudkovs, N., and Crane, M. St. J.: PCR-based assays for the fish pathogen aeromonas salrnonicida. II.Further evaluation and validation of three PCR primer sets with infected fish. Dis. Aquat. Org., 49: 139-144, 2002 https://doi.org/10.3354/dao049139
  7. Clark, D. J., Blake-Coleman, B. C., and Calder, M. R.: Principles and potential of piezo-electric transducers and acoustical techniques. in Biosensors : fundamentals and Application(Turner, A. P. F., Karube, I. and Wilson, G.S., eds.), Oxford University press. Oxford, pp 551-571, 1987
  8. Guilbault, G. G., Hock, B. and Schmid, R.: A Piezoelectric Immunobiosensor for Atrazine in Drinking Water. Biosensors Bioelectronics 7: 411-420, 1992 https://doi.org/10.1016/0956-5663(92)85040-H
  9. Horisberger, M. and Vauthey, M.: Labelling of colloidal gold with protein a quantitative study using $\beta$-lactoglobulin, Histochemistry 80(1):13-19, 1984 https://doi.org/10.1007/BF00492765
  10. Konig, B. and Gratzel, M.: Human granulocytes detected with a piezoimmunosensor, Anal. Letters. 26(11):2313, 1993 https://doi.org/10.1080/00032719308017472
  11. Konig, B. and Gratzel, M.: A novel immunosensor for herpes viruses, Anal. Chem. 66: 341, 1994 https://doi.org/10.1021/ac00075a005
  12. Kubota, S., Kaige, N., Miyazaki, T and Miyashita, T.: Histopathological studies of edwardsiellosis of Tilapia l. Natural infection. Vull. Fac. Fsih., Mie Univ., 9: 155-65, 1982
  13. Kusuda, R., Itami, T., Munekiyo, M. and Nakajima, H: Characteristics of all Edwardsiella sp. from an epizootic of cultured crimson sea breams. Bull. Jap, Soc. Scient. Fish., 43: 129-234, 1977 https://doi.org/10.2331/suisan.43.129
  14. Kusuda, R., Toyoshima, T., Iwamura, Y. and Sako, H.: Edwardsiella tarda from an epizootic of mullets (Mugil cephalus) in Okitsu Bay. Bull. Jap. Soc. Scient. Fish., 42: 271-275, 1976 https://doi.org/10.2331/suisan.42.271
  15. Lin, Z., Yip, C. M., Joseph, I. S. and Ward, M. D.: Operation of an Ultrasensitive 30-MHz Quartz Crystal Microbalance in Liquids, Anal. Chem. 65: 1546-1551, 1993 https://doi.org/10.1021/ac00059a011
  16. Matsumura, H. and Kleijn, J. M.: Admittance measurements on protein layers adsorbed at the Pt/solution interface : effect of d.c. potential and a.c. field. Colloids and surfaces. B, Biointerfaces, 277-282, 1993
  17. Meyer, F. P. and Bullock, G. L.: Edwatdsiella tarda, a new pathogen of channel catfish (Ictalurus punctatus) Appl. Microbiol. 25: 155-156, 1973
  18. Minunni, M., Mascini, M., Guilbault, G. G. and Hock, B.: The Quartz Crystal Microbalance as Biosensor. A Status Report.: Anal. Letters, 28: 749-764, 1995 https://doi.org/10.1080/00032719508001422
  19. Muramatsu, H., Dicks, J. M., Tamia, E. and Karube, I.: Piezoelectric Crystal Biosensor Modified with Protein A for Determination of Immunoglobulins. Analytical Chemistry, 59: 2760-2763, 1987 https://doi.org/10.1021/ac00150a007
  20. Prusak-Sochaczewski E, Luong, J. H. and Guilbault, G. G.: Development of a piezoelectric immunosensor for the detection of Salmonella typhimurium. Enzyme Microb Technol., 12(3): 173-177, 1990 https://doi.org/10.1016/0141-0229(90)90034-N
  21. Shons, A., Dorman and F. Najarian, J.: An immunospecific microbalance, J. Biomed. Mater. Res., 6: 565-570, 1972 https://doi.org/10.1002/jbm.820060608
  22. Skladal, P., Minunni, M., Mascini, M., Kolar, V., and Franek, M.: Characterization of MonoclonalAntibodies to 2.4A-Dichlorophenoxyacetic Acid Using A Piezoelectric Quartz- Crystal Microbalance in Solution. J. Immunol. Methods 176: 117-125, 1994 https://doi.org/10.1016/0022-1759(94)90356-5
  23. Steegborn, C. and Skladal, P.: Construction and characterisation of the direct piezoelectric immunosensor for atrazine operating in solution. Biosens. Bioelectron., 12:19-27, 1997 https://doi.org/10.1016/0956-5663(96)89086-5
  24. Uttenthaler, E., Kosslinger, C. and Drost, S.: Characterization of immobilization methods for African swine fever virus protein and antibodies with a piezoelectric immunosensor. Biosens. Bioelectron., 13: 1279-1286, 1998 https://doi.org/10.1016/S0956-5663(98)00089-X
  25. Vikholm, I., Viitala, T., Albers, W.M. and Peltonen, J.: Highly efficient immobilisation of antibody fragments to functionalised lipid monolayers, Biochim. Biophys. Acta., 1421: 39-52, 1999 https://doi.org/10.1016/S0005-2736(99)00112-1
  26. Wakabayashi, J. and Egusa, S.: Edwardsiella tarda (Paracolo- bacterium anguillimortiferum) associated with pond-cultured eel disease. Bull. Jap. Soc. Scient. Fish., 39:931-936, 1973 https://doi.org/10.2331/suisan.39.931
  27. Yu, L., Yuan, L., Feng, H. and Li, S.F.Y. : Determination of the bacterial pathogen Edwardsiella tarda in fish species by capillary electrophoresis with blue light-emitting diode-induced fluorescence. Electrophoresis, 25: 3139-3144, 2004 https://doi.org/10.1002/elps.200406018