Staphylococcus aureus 신속 검출을 위한 효소면역측정 스트립 센서

박소정; 김영기; So Jung Park; Young-Kee Kim

Applied Chemistry for Engineering, Vol.22, No.5, 522-525, October, 2011

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Staphylococcus aureus 신속 검출을 위한 효소면역측정 스트립 센서

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

박소정, 김영기^†

한경대학교 화학공학부

So Jung Park, and Young-Kee Kim^†

Department of Chemical Engineering, Hankyong National University, Anseong, Gyeonggi-Do 456-749, Korea

E-mail:

초록

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 × 10^(4)∼2.7 × 10^(7)CFU/mL 범위의 S. aureus 농도를 측정할 수 있었다.

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 × 10^(4)∼2.7 × 10^(7)CFU/mL can be detected by the colorimetric method within 30 min.

Keywords:Staphylococcus aureus;immuno-strip sensor;enzyme-linked immunosorbent assay (ELISA);immuno-chromatographic assay

[References]

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Yang L, Bashir R, Biotechnology Advances., 26, 135 (2008)
Park SG, Hwang YO, Jung JH, Lee KM, J. Food Hyg. Saf., 16, 159 (2001)
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Huang SH, Sensors and Actuators B., 127, 335 (2007)
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Lazcka O, Campo FJD, Munoz FX, Biosensors and Bioelectronics., 22, 1205 (2007)
Taylor AD, Ladd J, Yu Q, Homola SCJ, Jiang S, Biosensors and Bioelectronics., 22, 752 (2006)
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Paek SH, Lee CW, Lee CS, Yook SH, Korean J. Biotechnol. Bioeng., 11, 42 (1996)
Kim SH, Jung WS, Paek SH, Korean J. Biotechnol. Bioeng., 18, 190 (2003)
Ko S, Grant SA, Biosensors and Bioelectronics., 21, 1283 (2006)
Park JK, The Magazine of the Institute of Electronics Engineers of Korea., 28, 56 (2001)
Lee JH, Yang SS, Kim BW, Sim SJ, Chae H, Yoon HC, Colloids and Surfaces A: Physicochem. Eng. Aspects., 313, 509 (2008)
Ivnitski D, Abdel-Hamid I, Atanasov P, Wilkins E, Biosensor and Bioelectronics., 14, 599 (1999)
Chang CH, Kim SJ, Korean J. Vet. Res., 29, 503 (1989)
Yoon CH, Cho JH, Oh HI, Kim MJ, Lee CW, Choi JW, Peak SH, Biosensors and Bioelectronics., 19, 289 (2003)
Inoue K, Ferrante P, Hirano Y, Yasukawa T, Shiku H, Matsue T, Talanta., 73, 886 (2007)
Ho JA, Hsu HW, Huang MR, Analytical Biochemistry., 330, 342 (2004)
Varshney M, Li Y, Srinivasan B, Tung S, Sensors and Actuators B., 128, 99 (2007)
Han SM, Cho JH, Cho IH, Paek EH, Oh HB, Kim BS, Ryu C, Lee K, Kim YK, Paek SH, Analytica Chemica Acta., 587, 1 (2007)
Fernandez-Sanchez C, Mcneil CJ, Rawson K, Nilsson O, Leung HY, Gnanapragasam V, Journal of Immunological Methods., 307, 1 (2005)
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[1] Lee HJ, Korean Journal of Veterinary Public Health., 32, 81 (2008)

[2] Chang JM, Fang TJ, Food Microbiology., 24, 745 (2007)

[3] Yang L, Bashir R, Biotechnology Advances., 26, 135 (2008)

[4] Park SG, Hwang YO, Jung JH, Lee KM, J. Food Hyg. Saf., 16, 159 (2001)

[5] Lee JH, Song KY, Hyeon JY, Hwang IG, Kwak HS, Han JA, Chung YH, Seo KH, Korean J. Food Sci. Ani. Resour., 30, 410 (2010)

[6] Moon JY, Lee EJ, Kim YB, Journal of Bacteriology and Virology., 34, 91 (2004)

[7] Rall VLM, Vieira FP, Rall R, Vieitis RL, Fernandes Jr., Candeias JMG, Cardoso KFG, Araujo JP Jr., Veterinary Microbiology., 132, 408 (2008)

[8] Huang SH, Sensors and Actuators B., 127, 335 (2007)

[9] Alocilja EC, Radke SM, Biosensors and Bioelectronics., 18, 841 (2003)

[10] Lazcka O, Campo FJD, Munoz FX, Biosensors and Bioelectronics., 22, 1205 (2007)

[11] Taylor AD, Ladd J, Yu Q, Homola SCJ, Jiang S, Biosensors and Bioelectronics., 22, 752 (2006)

[12] Skottrup PD, Nicolaisen M, Justesen AF, Biosensors and Bioelectronics., 24, 339 (2008)

[13] Paek SH, Lee CW, Lee CS, Yook SH, Korean J. Biotechnol. Bioeng., 11, 42 (1996)

[14] Kim SH, Jung WS, Paek SH, Korean J. Biotechnol. Bioeng., 18, 190 (2003)

[15] Ko S, Grant SA, Biosensors and Bioelectronics., 21, 1283 (2006)

[16] Park JK, The Magazine of the Institute of Electronics Engineers of Korea., 28, 56 (2001)

[17] Lee JH, Yang SS, Kim BW, Sim SJ, Chae H, Yoon HC, Colloids and Surfaces A: Physicochem. Eng. Aspects., 313, 509 (2008)

[18] Ivnitski D, Abdel-Hamid I, Atanasov P, Wilkins E, Biosensor and Bioelectronics., 14, 599 (1999)

[19] Chang CH, Kim SJ, Korean J. Vet. Res., 29, 503 (1989)

[20] Yoon CH, Cho JH, Oh HI, Kim MJ, Lee CW, Choi JW, Peak SH, Biosensors and Bioelectronics., 19, 289 (2003)

[21] Inoue K, Ferrante P, Hirano Y, Yasukawa T, Shiku H, Matsue T, Talanta., 73, 886 (2007)

[22] Ho JA, Hsu HW, Huang MR, Analytical Biochemistry., 330, 342 (2004)

[23] Varshney M, Li Y, Srinivasan B, Tung S, Sensors and Actuators B., 128, 99 (2007)

[24] Han SM, Cho JH, Cho IH, Paek EH, Oh HB, Kim BS, Ryu C, Lee K, Kim YK, Paek SH, Analytica Chemica Acta., 587, 1 (2007)

[25] Fernandez-Sanchez C, Mcneil CJ, Rawson K, Nilsson O, Leung HY, Gnanapragasam V, Journal of Immunological Methods., 307, 1 (2005)

[26] Choi JG, Shin WB, Je JH, Kim JY, Lee KH, Kim MG, Ha SD, Kim KS, Kim KY, Kim CH, Chung DH, Korean J. Food Sci. Technol., 39, 299 (2007)